text:
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey.
ABSTRACT Lateral flow (immuno)assays are currently used for qualitative, semiquantitative and to some extent quantitative monitoring in resource-poor or non-laboratory environments. Applications include tests on pathogens, drugs, hormones and metabolites in biomedical, phytosanitary, veterinary, feed/food and environmental settings. We describe principles of current formats, applications, limitations and perspectives for quantitative monitoring. We illustrate the potentials and limitations of analysis with lateral flow (immuno)assays using a literature survey and a SWOT analysis (acronym for "strengths, weaknesses, opportunities, threats"). Articles referred to in this survey were searched for on MEDLINE, Scopus and in references of reviewed papers. Search terms included "immunochromatography", "sol particle immunoassay", "lateral flow immunoassay" and "dipstick assay".
- Jinsu Kim, Meena Adhikari, Sagar Dhamane, Anna E V Hagström, Katerina Kourentzi, Ulrich Strych, Richard C Willson, Jacinta C Conrad[Show abstract] [Hide abstract]
ABSTRACT: We demonstrated a lateral flow immunoassay (LFA) for detection of viruses using fluorescently-labeled M13 bacteriophage as reporters and single-reporter counting as the readout. AviTag-biotinylated M13 phage were functionalized with antibodies using avidin-biotin conjugation and fluorescently labeled with AlexaFluor 555. Individual phage bound to target viruses (here MS2 as a model) captured on an LFA membrane strip were imaged using epi-fluorescence microscopy. Using automated image processing, we counted the number of bound phage in micrographs as a function of target concentration. The resultant assay was more sensitive than enzyme-linked immunosorbent assays and traditional colloidal-gold nanoparticle LFAs for direct detection of viruses.ACS Applied Materials & Interfaces 01/2015; 7(4). · 5.90 Impact Factor - SourceAvailable from: PubMed Central[Show abstract] [Hide abstract]
ABSTRACT: We demonstrate a fluorescence lateral flow system that has excellent sensitivity and wide dynamic range. The illumination system utilizes an LED, plastic lenses and plastic and colored glass filters for the excitation and emission light. Images are collected on an iPhone 4. Several fluorescent dyes with long Stokes shifts were evaluated for their signal and nonspecific binding in lateral flow. A wide range of values for the ratio of signal to nonspecific binding was found, from 50 for R-phycoerythrin (R-PE) to 0.15 for Brilliant Violet 605. The long Stokes shift of R-PE allowed the use of inexpensive plastic filters rather than costly interference filters to block the LED light. Fluorescence detection with R-PE and absorbance detection with colloidal gold were directly compared in lateral flow using biotinylated bovine serum albumen (BSA) as the analyte. Fluorescence provided linear data over a range of 0.4-4,000 ng/mL with a 1,000-fold signal change while colloidal gold provided non-linear data over a range of 16-4,000 ng/mL with a 10-fold signal change. A comparison using human chorionic gonadotropin (hCG) as the analyte showed a similar advantage in the fluorescent system. We believe our inexpensive yet high-performance platform will be useful for providing quantitative and sensitive detection in a point-of-care setting.Biosensors. 01/2013; 3(4):360-73. -
Article: Dual-layered and double-targeted nanogold based lateral flow immunoassay for influenza virus
Microchimica Acta 01/2014; 182(1-2):85-93. · 3.72 Impact Factor
Page 1
Lateral flow (immuno)assay: its strengths, weaknesses,
opportunities and threats. A literature survey
Geertruida A. Posthuma-Trumpie & Jakob Korf &
Aart van Amerongen
Received: 21 May 2008 /Revised: 1 July 2008 /Accepted: 3 July 2008 / Published online: 13 August 2008
# The Author(s) 2008
Abstract Lateral flow (immuno)assays are currently used
for qualitative, semiquantitative and to some extent quan-
titative monitoring in resource-poor or non-laboratory
environments. Applications include tests on pathogens,
drugs, hormones and metabolites in biomedical, phytosani-
tary, veterinary, feed/food and environmental settings. We
describe principles of current formats, applications, limi-
tations and perspectives for quantitative monitoring. We
illustrate the potentials and limitations of analysis with
lateral flow (immuno)assays using a literature survey and a
SWOT analysis (acronym for "strengths, weaknesses,
opportunities, threats"). Articles referred to in this survey
were searched for on MEDLINE, Scopus and in references
of reviewed papers. Search terms included "immunochro-
matography", "sol particle immunoassay", "lateral flow
immunoassay" and "dipstick assay".
Keywords Biomonitoring.Clinicalchemistry.
This study reports on the lateral flow (immuno)assay/
immunochromatography technique. Originally the tech-
nique was called "sol particle immunoassay" (SPIA) [1].
Lateral flow assays (LFAs), i.e. prefabricated strips of a
carrier material containing dry reagents that are activated by
applying the fluid sample, are important for diagnostic
purposes, e.g. to ascertain pregnancy, failure of internal
organs (e.g. heart attack, renal failure or diabetes), infection
or contamination with specific pathogens including biowar-
fare agents, presence of toxic compounds in food, feed or
the environment and abuse of (illicit) drugs. These assays
are often incorrectly referred to as "dipsticks". Real dipstick
assays are based on the immunoblotting principle and do
not rely on lateral fluid flow through a membrane. In this
survey lateral flow (immuno)assays are evaluated and,
therefore, dipsticks that are lateral flow assays will be
named LFAs. They are especially designed for single use at
point of care/need, i.e. outside the laboratory. Applications
are often designed where an on/off signal is sufficient. The
best-known application is the pregnancy test [1, 2]. Results
usually come within 10–20 min. The current generation of
LFAs has high sensitivity, selectivity and ease of use. In
most cases sensitivity and selectivity are achieved by
combining miniaturised thin-layer chromatography, the
use of analyte-specific antibodies or DNA/RNA specific
sequences, and by labelling the analyte or the recognition
The assay format in lateral flow immunoassay (LFIA) is
comparable to that in enzyme immunoassay, although it
does not replace that assay format. In fact, in the first
reports on LFIAs use of the same components as in enzyme
immunoassays was reported, immobilising the antibody on
Anal Bioanal Chem (2009) 393:569–582
DOI 10.1007/s00216-008-2287-2
G. A. Posthuma-Trumpie:J. Korf
Department of Psychiatry,
University Medical Centre Groningen (UMCG),
University of Groningen,
Hanzeplein 1, P.O. Box 30.001, 9700 RB Groningen,
The Netherlands
A. van Amerongen (*)
Agrotechnology and Food Sciences Group,
Wageningen University and Research Centre,
Bornsesteeg 59, P.O. Box 17, 6700 AA Wageningen,
The Netherlands
e-mail: firstname.lastname@example.com
Page 2
a chromatographic paper strip [3]. In doing so, advantage
could be made of the chromatographic principle in addition
to the immunological recognition system. The first labels
were the enzymes used for enzyme immunoassays [4], but
they were replaced soon by particulate labels [1]. Nowa-
days when a new method is presented for LFIAs, the
performance of the new system is often referenced to the
performance of enzyme immunoassays [5–8].
Lateral flow (immuno)assay is a technology that is
currently widely applied [5–16]; however, to our knowl-
edge no in-depth survey of recent literature is available.
Here we present a survey of development reports presented
in the scientific literature, mainly during the last decade,
where a selection of those applications is mentioned that are
real one-step dry reagent assays with coloured or lumines-
cent response. A discussion on the strengths of, weaknesses
of, opportunities for and threats to this technique is
included as well.
Principles of lateral flow (immuno)assays
Essential in the current lateral flow (immuno)assay (LFA)
formats is the movement of a liquid sample, or its extract
containing the analyte of interest, along a strip of polymeric
material thereby passing various zones where molecules
have been attached that exert more or less specific
interactions with the analyte. A typical LFA format consists
of a surface layer to carry the sample from the sample
application pad via the conjugate release pad along the strip
encountering the detection zone up to the absorbent pad.
The various elements will be discussed in more detail
below; see also Fig. 1.
The membrane is often thin and fragile, so it is attached
to a plastic or nylon basic layer to allow cutting and
handling. In addition, robustness is achieved by housing the
strips in a plastic holder, where only the sample application
window and a reading window are exposed. Current
membrane strips are produced from nitrocellulose, nylon,
polyethersulfone, polyethylene or fused silica.
At one end of the strip a sample application pad is
provided. The sample application pad is usually made of
cellulose or cross-linked silica.
In close contact with the strip material and the sample
application pad is the conjugate release pad, made of cross-
linked silica. Labelled analyte or recognition element(s)
(depending on the application) are dried on this pad and
after addition of the sample, this material will interact with
the fluid flow; specific interactions will be initiated here
and will continue during the chromatographic process.
Alternatively, the labelled analyte or recognition element(s)
are dried in a reaction tube and the sample plus strip are
added to the tube [6] (Fig. 2).
Labels are made of coloured or fluorescent nanoparticles
with sizes of 15–800 nm, allowing an unobstructed flow
through the membrane. They are often made of colloidal
gold [1, 17–20] or latex [21], less often selenium [22],
carbon [2, 23] or liposomes [24–29] are used. Latex
nanoparticles are coloured [21] for optical detection. In
liposomes coloured [21], fluorescent [27] or bioluminescent
[29] dyes can be incorporated, allowing visualisation, and,
when applicable, quantitation of the response. The newest
labels include quantum dots [30] and upconverting phos-
phor technology [9, 31].
At least two lines are sprayed on the strip: a test line and
a control line. At the test line the recognition of the sample
analyte and the reporter will result in the required response.
Fig. 1 Lateral flow immunoassay (LFIA) in sandwich format. Nano-
particle labelled antianalyte antibody 1 is dried at the conjugate release
pad. Antianalyte antibody 2 is sprayed at the test line (T). Antispecies
immunoglobulin G is sprayed at the control line (C). Sample flows
from the sample pad to the conjugate pad and into the membrane.
Strips are mounted in a device for protection and easier handling
Fig. 2 Lateral flow (immuno)assay tube format; conjugate is
dehydrated in a test tube. Tube and strip are stored in a sealed
aluminium pouch and a desiccant. To run the test, sample (and buffer)
are pipetted into the test tube, conjugate is dissolved and the strip is
inserted. Response at the test line (T) is dependent on the analyte
concentration; response at the control line (C) indicates a proper flow
through the membrane
570G. Posthuma-Trumpie et al.
Page 3
A response at the control line confirms a proper flow of the
liquid through the strip. More test lines can be applied
allowing for multianalyte testing [32–36] or for semiquan-
titative evaluation of the response [37, 38].
The liquid moves because of the capillary force of the
strip material, but to maintain a flow an absorbent pad is
attached at the distal side of the strip. This absorbent pad
will wick the liquid to the end of the strip, thus maintaining
the flow.
When exclusively antibodies are used as recognition
elements, the tests are called "lateral flow immunoassays"
(LFIA). A combination of antigen-antibody interaction and
specific tagged doubled-stranded amplicon (ds-amplicon)
detection after PCR is also possible, and is called "nucleic
acid lateral flow immunoassay" (NALFIA) [39, 40, 42, 69].
Specific nucleic acid hybridisation of amplicons with
immobilised complementary probes is another option and
is called "nucleic acid lateral flow assay" (NALF) [16, 41].
A selection of published LFA applications with the method
applied and the sensitivity is presented in Tables 1, 2, 3 and
4; these were mainly published in the last decade. The
examples have been grouped into four categories: infections
(Table 1), metabolic disorders (Table 2), toxic compounds
(Table 3), and miscellaneous applications (Table 4).
Lateral flow immunoassay
The LFIA set-up is designed to confirm the presence or
absence of an (un)wanted compound; the analyte under
consideration can be this compound, or a metabolic
derivative. Antibodies to the analyte are used for recogni-
tion. Several formats have been described for LFIAs. These
formats are chosen depending on the analyte. When the
analyte is of low molecular mass and has only one epitope,
i.e. a hapten, the format is restricted to the competitive
design. Two layouts are possible: (1) antibody is sprayed at
the test line, a mixture of sample analyte and labelled
analyte is applied at the conjugate pad and the sample
analyte and labelled analyte compete for binding sites on
the antibody at the test line [68]; (2) an analyte-protein
conjugate is sprayed at the test line, and a mixture of
labelled antibody and sample analyte is applied at the
conjugate pad, giving the sample analyte a head start for
binding to the antibody [13]. The preferred layout is
dependent on the particular application. In the competitive
LFIA format the response is negatively correlated to the
analyte concentration (i.e. more analyte present, less signal;
no analyte gives the highest signal). A typical scheme of
the competitive design and the response using option 2 is
depicted in Fig. 3 (top).
For analytes with more than one epitope, the sandwich
format is applicable. In this format the test line is prepared
using one analyte-specific antibody. The conjugate release
pad contains a second, labelled, antianalyte antibody.
Analyte in the sample will bind during the initial chromato-
graphic process to the second antibody. The free epitope can
bind to the immobilised antibody at the test line [65]. The
response is directly proportional to the amount of analyte in
the sample; a scheme is depicted in Fig. 3 (bottom).
Nucleic acid lateral flow (immuno)assay
The NALFIA and the NALF set-ups are usually designed
for testing the presence or absence of pathogens in food,
feed or the environment. In the NALFIA set-up the analyte
is an amplified double-stranded nucleic acid sequence (ds-
amplicon) specific of the organism using primers with two
different tags; recognition of the analyte is done by binding
to a tag-specific antibody [39, 40, 42, 69]. In a typical
layout developed for the detection of pathogenic bacteria
the nucleic acid was amplified using PCR with two tagged
primers (Fig. 4a). A ds-amplicon was obtained with one
strand labelled with biotin and the other strand labelled
with, e.g., fluorescein isothiocyanate or digoxigenine. A
solution of antibodies raised against the tag was sprayed at
the test line. The biotin will bind to the avidin-labelled
nanoparticles and the other tag will bind to the antitag
antibody, resulting in the coloured signal [39]. The response
is directly proportional to the amount of analyte.
For the NALF set-up several formats have been
published: (1) nanoparticle-labelled reporter probe and
biotin-labelled immobilised capture probe via avidin,
single-stranded amplicon (ss-amplicon) hybridises with
complementary reporter and capture probes (Fig. 4b) [16];
(2) nanoparticle-labelled reporter probe and bovine serum
albumin labelled capture probe, immobilised through
passive adsorption, ss-amplicon hybridises with comple-
mentary reporter and capture probes (Fig. 4c) [70]; (3)
nanoparticle-labelled reporter probe; capture probe is
immobilised at the test line through passive adsorption,
ss-amplicon hybridises with complementary reporter and
capture probes (Fig. 4d) [70]. The response is directly
proportional to the amount of analyte [16].
Critical parameters and sensitivity of the assay
Membrane material
The membrane used as strip material is probably the single
most important part of the test system. Most often the strip
material is made of nitrocellulose [12, 19, 23, 33, 46, 58,
63, 71, 72], available from Whatman, Pierce, Advanced
Microdevices (India) or Millipore. Several other polymeric
materials are also available, e.g. nylon [73] (Millipore, Pall
Biosciences), polyethersulfone [15, 16] (Pall Biosciences or
Nalgene-Nunc), polyethylene [51] (Porex) or fused silica
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey 571
Page 4
(Fusion5, Whatman), but these are used less often. A slight
tendency to use polyethersulfone material was observed for
the NALF format [15, 16]. Important are not only the
capillary forces of the carrier material, but also the ease of
binding and immobilising proteins necessary for sequential
selection, reaction and detection.
For nitrocellulose, various pore sizes are available,
varying from 0.05 to about 12 μm. However, the pores
are not randomly distributed because of the manufacturing
process, and a better description of the strip material should
be capillary flow time, i.e., the time for the fluid front to
travel into the strip material and is expressed as seconds per
centimetre. First of all, the pore size and the material are
important for the transport of the label [74]. A second
requirement is the speed with which the complex of the
sample analyte and the label are transported through the
Table 1 Overview of lateral flow (immuno)assays: detection of infectious agents
AnalyteMethod ApplicationSensitivitya
Specific RNA after isothermal
Polystyrene dyed microsphere
labelled specific
Colloidal gold labelled
oligo dT strands
Colloidal dye (palanyl red)
labelled monoclonal antihuman
IgM antibody
Colloidal gold labelled
primary antibody
Dye-entrapped and
Dye-entrapped liposome
DNA probe
Detection of Bacillus
2 B. anthracis cells[43]
PCR amplified product with
dA tail of 23S ribosomal RNA
Brucella-specific IgM
antibodies in serum
Bacterial infections
in arthroplasty
10 cells of S. aureus [15]
93% positives [44]
Citrus tristeza virus in
extracts from leaves or fruits
Cryptosporidium species
oocytes in water
Detection of virus infection90% (leaves)a;
97% (fruits)a
1 oocyst in 10 μL
Contamination of
4 dengue serotypes after
isothermal nucleic acid
based amplification in serum
Human or animal IgG
in mosquito bloodmeal
Detection of virus infection 50–50,000 copies of
RNA molecules in
10 μg L−1
Dye-labelled anti-IgG antibodiesIgG (human or chicken) to detect
different host sources of blood
meal in insects
Multianalyte detection of influenza
A, influenza B and respiratory
syncytial virus
Influenza A, influenza B and
respiratory syncytial virus
Raman signature labelled
antigen measures Raman
Colloidal dye labelled
monoclonal antihuman IgM
Colloidal gold labelled primary
Colloidal gold labelled antibody
Leptospira-specific IgM
in serum
87% overall
Rotavirus in bovine faeces Detection of Rotavirus
Detection of virus infections
70% of positives[17]
Satsuma dwarf virus in
fruit extracts
Antibody to Schistosoma
japonicum in serum
Blue colloidal dye labelled egg
Schistosomiasis97% positives in
acute form;
94% in chronic
0.2 μg L−1
Schistosoma circulating
cathodic antigen in urine
Streptococcus pneumoniae
DNA in a complex matrix
(fish sperm)
Antibodies to Treponema
pallidum and hepatitis B
antigen in serum
Antibody to Trichinella in
swine serum
Colloidal carbon labelled
primary antibody
Upconverting phosphor-labelled
species-specific DNA sequence
Streptococcus pneumoniae
1 ng of AluI-
restricted S.
pneumonia DNA
5 μg L−1hepatitis
B antigen
Colloidal gold and
oligonucleotide labelled
Colloidal gold labelled antigen
Treponema pallidum;
hepatitis B infection
Trichinellosis100% of positives[19]
IgG immunogobulin G, IgM immunogobulin M
aPercentage of positives compared to the "gold standard"
572G. Posthuma-Trumpie et al.
Page 5
membrane allowing optimal reaction time. These parame-
ters are also important when the sample has a high viscosity
or contains fat globules, e.g. milk.
Usually nitrocellulose material to prepare the strips can
be stored at ambient temperature and humidity; however, at
low humidity, handling of the material can be difficult
owing to accumulation of static electricity.
For polyethylene, the pore size can be controlled
precisely, and when there is need for such properties, i.e.
better reproducibility during the assembly and consequently
better reproducibility during the tests, using this strip
material is an option [51]. It might be noted here that
larger pore sizes will widen the test line, and the sensitivity
of the test will decrease, because faint lines can be missed.
However, publications describing a comparison of the
performance of different strip materials could not be found.
Material of the sample pad
The sample pad can be used to perform multiple tasks; the
most important is the promotion and even distribution of
the sample to the conjugate pad. The sample pad can also
be impregnated with, e.g. proteins, detergents, viscosity
enhancers and buffer salts to influence the flow rate of the
sample. The aim of such additions can be to increase the
sample viscosity, to increase the reaction time at the
conjugate pad or even to chemically modify the sample
for binding at the test line. The pores in the sample pad can
be symmetrically, homogeneously or inhomogeneously
distributed, or can be asymmetrically distributed, providing
an initial filter to remove coarse material, e.g. whole cells.
Asymmetrical pore size distribution can be obtained from,
e.g., Pall Ahlstrom (Cytosep) [6, 47, 55], or from Advanced
Microdevices (India) [63].
The material of the sample pad is often chosen in
advance in accordance with the aim of the test and the
properties of the sample. Optimisation is usually not
Material of the absorbent pad
The primary function of the absorbent pad is to wick the
fluid through the membrane. When an absorbent pad is
used, the amount of sample can be increased, resulting in an
increased sensitivity. Most often cellulose filters are used.
Material and size of the label
As material of the label nowadays colloidal gold is used
most often [1, 5, 7, 10, 15, 18–20, 33, 34, 51, 53, 54, 57,
Table 2 Overview of lateral flow (immuno)assays: detection of metabolic disorders
Analyte MethodApplicationDetection limitReference
Albumin in serum/whole
Fluorescent dye labelled
antibody or fluorescent dye
labelled antigen
Fluorescent dye labelled
Albumin as a biomarker
for diabetes
12.2–60 g L−1
C-reactive protein in
whole blood
C-reactive protein; heart-
type fatty acid binding
protein in plasma
Lipoprotein A in serum,
plasma or blood
Albumin in urine
C-reactive protein as a potential
risk predictor for future
cardiovascular diseases
C-reactive protein and heart-type fatty
acid binding protein as biomarkers of
cardiovascular diseases
Lipoprotein A as biomarker
for atherosclerosis
0.133 mg L−1
Colloidal gold labelled
primary antibody
2 mg L−1C-reactive protein;
5 μg L−1heart-type fatty
acid binding protein
<40 mg L−1lipoprotein A
Selenium-labelled antigen
Alkaline colour reaction to
tetrabromphenol blue
Eu(III) chelate microparticles
conjugated to primary
Gold-labelled secondary
Colloidal gold labelled
primary antibody and biotin-
labelled primary antibody
Europium- or biotin-labelled
Eosinophil protein X;
neutrophil lipocalin in
whole blood
Free and total prostate-
specific antigen
Human serum albumin
in urine
Point of care for neutrophil,
eosinophil and leucocyte counts
0.082 g L−1for eosinophil
protein X; 0.05 g L−1for
neutrophil lipocalin
1 μg L−1prostate-specific
30 mg L−1
Screening on prostate-specific
Detection of renal disease and
its progression
Total PAPP-A and
PAPP-A complexed with
PAPP-A as a biomarker for
acute coronary syndromes
0.18 mIU L−1for total
PAPP-A and 0.23 mIU L−1
for PAPP-A/proMBP
PPV Positive predictive value of indicated metabolic disorder, PAPP-A pregnancy-associated plasma protein A, proMBP pro form of eosinophil
major basic protein
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey573
Page 6
61, 66, 67, 71, 75–77]. A runner-up is the use of coloured
latex particles [21, 76, 78]. The use of other coloured
nanoparticles was mentioned as well. Among those are
carbon [2, 23] and selenium [22]. Chemiluminescent and
fluorescent nanoparticles are used less often [9, 16, 58,
79]. An important requirement for the nanoparticle labels is
their colloidal stability in solution. Among others, this can
be achieved upon coating the binding proteins on the
surface of the nanoparticle. Electrical charge can be
generated by choosing the right conditions (i.e. buffer
composition, buffer strength, pH). A stable suspension
ensures a uniform distribution of the conjugate when
preparing the conjugate pad. The size of the gold nano-
particles can be controlled during manufacturing. Several
publications report on a comparison of the size of the
nanoparticles [68, 74, 77]. Better sensitivity using larger
particles was observed; however, the stability of the colloid
was diminished with particles over 40 nm [68]. This
problem was not mentioned in [77], where an optimum
was found using gold nanoparticles of 80 nm. Liposome
colloids can be instable as well, so in a typical application,
the liposomes were immobilised and the label ruptured the
liposomes, releasing an electroactive compound in a
concentration-dependent way [80].
As was the case for strip material, a comparison of
sensitivities using different labels could not be found.
Recognition element
The sensitivity of the LFIA is to a large extent dependent
on the affinity of the specific antibody. Specific antibodies
(primary antibodies) can be labelled to provide a response;
however, a high concentration of, often expensive, material
is necessary. When secondary antispecies antibodies (often
Table 3 Overview of lateral flow (immuno)assays: detection of toxic compounds
AnalyteMethodApplicationDetection limit References
Aflatoxin B1in pig feed
Botulinum neurotoxin D
in horse faeces
Carbaryl and endosulfan
in extracts of cereals and
(Dihydro)streptomycin in raw
Colloidal gold labelled primary
Colloidal gold labelled primary
Colloidal gold or HRP labelled
primary antibody
Contamination with
aflatoxin B1
Screening on Botulinum
Pesticide residues of
carbaryl and endosulfan
2 μg kg−1
50 ng L−1
100 μg L−1for carbaryl;
10 μg L−1for endosulfan
Colloidal gold labelled primary
Antibiotics residues of
25 μg kg−1for
streptomycin; 50 μg kg−1
for dihydrostreptomycin
1.0 μg L−1
Fumonisin B1 in extracts of
cereals and peanuts
Glycyrrhizic acid in extracts of
roots, leaves or stems of
liquorice plant samples
Medroxy-progesterone acetate
in extractions of swine liver or
Microcystin-LR in freshwater
Colloidal gold labelled primary
Colloidal gold labelled primary
Contamination with
mycotoxin fumonisin B1
Determination of
glycyrrhizic acid
Residue of analyte
20–50 μg L−1
Colloidal gold labelled primary
10 mg kg−1
Biotin-fluorescein-labelled analyte
or antianalyte biotin-fluorescein-
labelled primary antibody
Blue latex microsphere labelled
primary antibody
Colloidal gold labelled primary
Colloidal carbon labelled secondary
Colloidal gold labelled antibody
Determination of
microcystin presence
50 ng L−1or 100 ng L−1
Nicarbazin residues in poultry
Sulfadimidine in urine or milk
Residue of pesticide
Residue of antibiotic
Residue of antibiotic
Residue of antibiotics
2 mg kg−1
10 μg L−1in urine; 10 μg
L−1in fresh bovine milk
6.3 μg L−1in diluted
(1/10) urine
10–40 μg L−1
Sulfametazine in urine
Generic test on Sulfonamides
in eggs and chicken muscle
Verotoxigenic Escherichia coli
in raw milk, minced beef,
apple juice and salami
Colloidal carbon labelled primary
Single analyte and
multianalyte verotoxigenic
Escherichia coli detection
100% positivesa
[11, 60]
HRP horseradish peroxidase
aPercentage of positives compared to the "gold standard"
574G. Posthuma-Trumpie et al.
Page 7
less expensive) are labelled, the primary antibodies can be
titrated for optimal response [13]. Moreover, the affinity of
the primary antibodies might decrease upon conjugation to
the label, diminishing the sensitivity. This phenomenon is
comparable to immobilisation of an enzyme in a reactor,
shown, e.g., in [81]. Steric hindrance might hamper the
antigen-antibody interaction as well. The dependency of the
affinity on labelling primary antibody cannot be predicted;
Fig. 3 Difference in layout and signal generation between competi-
tive LFIA and sandwich LFIA. In the competitive format the response
is negatively correlated to the amount of analyte in the sample (top).
In the sandwich format the response is directly proportional to the
amount of analyte in the sample (bottom)
Table 4 Overview of lateral flow (immuno)assays: miscellaneous applications
AnalyteMethod ApplicationDetection limitReferences
protective IgG in
serum or whole
Bovine IgG in dried
porcine plasma
Cortisol in blood/
Fungal α-amylase in
hGC in urine
Colloidal gold labelled
streptavidin with analyte
Confirmation of previous infection or
3 mg L−1IgG in serum;
∼14 mg L−1IgG in whole
blood; 100% positivesa
Colloidal gold labelled
primary antibody
Colloidal gold labelled
primary antibody
Colloidal carbon labelled
primary antibody
Bovine IgG as marker of plasma
Quantitation of cortisol concentration
0.01% v/v[62]
3.5 μg L−1in plasma[63]
Fungal α-amylase contamination at
the workplace
1–10 μg L−1
[6, 64]
Colloidal carbon labelled
primary antibody
Colloidal gold labelled
primary or secondary
Uniformly blue dyed
polystyrene microsphere
labelled primary antibody
Colloidal gold labelled
analyte-ovalbumin conjugate
Determination of presence of hCG
(pregnancy test)
Determination of presence of hCG
(pregnancy test) or TPSA for
screening for prostate cancer
Confirmation of pregnancy
10 mIU mL−1
hCG or TPSA1 ng L−1hCG in urine; 10 ng
L−1hCG in serum; 0.2 μg L−1
TPSA in serum
5 μg L−1
Oestrone sulphate in
mares' urine
Progesterone in
bovine milk
Determination of the reproductive
status of dairy animals
5 μg L−1
hGC human chorionic gonadotropin, TPSA total prostate-specific antigen
aPercentage of positives compared to the "gold standard"
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey575
Page 8
however, comparison of labelling primary or secondary
antibodies on the sensitivity of the assay could not be
For the nuclei acid assays, the availability of good
primers during the amplification process is indispensable.
In the NALFIA format the recognition is done by antibody
tag-label interaction and the biotin-avidin reaction. In the
NALF format complementary probes have to be hybridised
to the amplicons. This hybridisation is often done before
the chromatographic process. It might be noted here that
hybridisation usually takes more time than antibody tag-
label interaction, favouring NALFIA over NALF.
Position of the test line
The position of the test line is important for the perfor-
mance of the assay. More interaction time is available when
the test line is farther downstream from the sample
application side of the strip. An optimum is usually
obtained, as the speed of movement of the fluid decreases
with distance.
Usually a first step in the development of the assay is the
optimisation of the concentration of the label and the
recognition element. The second step is the optimal position
of the test line. Both are aimed at optimal sensitivity of the
assay. Strip material, the label and the detection strategy are
usually the personal choice of the research group developing
the assay. However, often the development of a new test is
aimed at circumventing existing patents.
SWOT analysis
A SWOT (acronym for "strengths, weaknesses, opportuni-
ties and threats") analysis of the lateral flow (immuno)assay
format is discussed here. An overview of strengths and
weaknesses, internal to the format, is given in Table 5. An
overview of opportunities and threats, presented by the
external environment, is given in Table 6.
Many successful LFAs are mentioned and all are designed
for use at point of care/need: the physician's examination
room, emergency ward in general hospitals, tests on food or
feed for toxic compounds, presence or absence of infectious
diseases in medical or phytopathological settings or adulter-
ation with undesirable components in food/feed industries.
Also for tests related to biowarfare, law enforcement and
forensics, this format is preferred regularly. However,
regulatory bodies often require an independent test to be
performed on samples that test positive. In this case LFA
tests can be used for primary screening at point of care/need.
Fig. 4 A number of possible formats for nucleic acid lateral flow
assays; detail at the test line (not to scale). a Nucleic acid lateral flow
immunoassay test format: nanoparticle-labelled avidin is used as
reporter and two tagged primers (e.g. biotin and fluorescein
isothiocyanate, FITC) label the double-stranded amplicon (ds-ampli-
con). Biotin binds to avidin; and anti-FITC antibody, sprayed at the
test line, binds to FITC. b Nucleic acid lateral flow test format:
nanoparticle-labelled reporter probe and biotin-labelled immobilised
capture probe, single-stranded amplicon (ss-amplicon) hybridises with
reporter and capture probes. c Nucleic acid lateral flow test format:
nanoparticle-labelled reporter probe and bovine serum albumin
labelled capture probe, immobilised through passive adsorption, ss-
amplicon hybridises with reporter and capture probes. d Nucleic acid
lateral flow test format: nanoparticle-labelled reporter probe; capture
probe is immobilised at the test line through passive adsorption, ss-
amplicon hybridises with reporter and capture probes
576G. Posthuma-Trumpie et al.
Page 9
Especially in third-world countries these tests are used
for biomedical purposes, since there is no need to
refrigerate the strips prepared. Moreover, visual interpreta-
tion of the results is often satisfactory. Owing to their
extended shelf life, large batches can be prepared, dimin-
ishing the variation between batches.
LFAs are designed for disposable single use and no
contamination with previously tested sample will occur.
The weaknesses will be discussed in more detail and, if
possible, improvements will be proposed, especially with
respect to quantitative results.
LFAs are a one-step assay, no washing step is possible.
Preblocking of the strips is not advisable. Nitrocellulose
immunostrips are prepared with well-balanced hydropho-
bicity and wetting properties. Washing or pretest blocking
will destroy these properties. However, in several publica-
tions preblocking the strips was mentioned [38, 68, 74].
Tests are developed aiming at on/off results. When a more
semiquantitative result is required, simple evaluation by
spraying a series of lines of capture reagent with increasing
concentration (comparable to a bar code) is sometimes
sufficient [37, 38]. A more extended format uses evaluation
of the signal by a measuring device. This can be a simple
hand-held reflectometer [24, 82, 83], but when a more
precise evaluation is required, the signal can be digitised
using a flatbed scanner or a CCD camera and dedicated
software [23, 39, 65]; however, costs and analysis time will
increase, although there is a trend in reduction of hardware
costs due to miniaturisation. In contrast, using a reader
diminishes subjective user interpretation, enables the storage
of data and is often more sensitive than visual interpretation.
In the competitive format the response is negatively
correlated to the concentration. Especially when it is used
Table 6 SWOT analysis of lateral flow assay: external factors
New applications at point of care/need
Application on other biomatrices: tears, saliva, sweat
Better reproducibility using automation of manufacturing, sample
and reagent addition, drying time and read-out of results
Miniaturisation of the LFIA strips
GC-MS or LC-MS in an automated format
Automated enzyme innunoassays
Apoenzyme reactivation immunological system
Immunosensor formats (e.g. Biacore, grating coupler, total internal
reflectance fluorescence, magneto-biosensor, Young interferometer sensor)
Microparticle immunoassays (e.g. Luminex® and related brands)
Lab-on-a-chip technology
Concentration of lateral flow (device) patents at one commercial enterprise
(Inverness Medical Innovations)
Development of high-throughput LFIA assays
Sweeping of the market at existing and upcoming economies
New applications can be ahead of other technologies owing to
faster development
GC-MS gas chromatography-mass spectrometry, LC-MS liquid chromatography-mass spectrometry, LFIA lateral flow immunoassay
Table 5 SWOT analysis of lateral flow assay: internal factors
Strengths Weaknesses
One-step assay, no washing step necessary
Fast and low cost, low sample volume
Reduced development time brings applications faster to the market
One-step assay, no washing step possible
Qualitative or semiquantitative results
Response is negatively correlated to concentration in competitive
Imprecise sample volume reduces precision
Restriction on total volume in test gives a limit on sensitivity
In one-step format no possibility to enhance the response by enzyme
Good antibody preparation or hybridisation nucleic acid sequence
Usually designed for individual tests, not for high-throughput
Obstruction of pores due to matrix components
Simple test procedure
Applications at point of care/need
Sensitive for proteins, haptens and nucleic acid amplicons
Versatile format
Individual tests or sometimes array format for batchwise mid-throughput
Prolonged shelf life without the need to refrigerate, larger batches can be
prepared in advance
Qualitative (on/off) or semiquantitative result
Sample pretreatment is often not necessary when the sample is a fluid
Immunostrips usually not manufactured for the purpose
Sample pretreatment is obligatory when the sample is not a fluid
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey 577
Page 10
by untrained persons, this might cause ambiguities. Chang-
ing the format by using an "immunothreshold" can
overcome this problem [63]. In this format two lines are
sprayed on the immunostrip, line 1 is the analyte-protein
conjugate and line 2 is the secondary antibody. Primary
antibodies conjugated to colloidal nanoparticles are applied
on the conjugate release pad. When no analyte is present in
the sample, all antibodies will bind to the immobilised
analyte at line 1; however, when analyte is present, primary
antibodies will bind to line 1 and/or line 2 in an analyte-
concentration-dependent way, and the intensity of the latter
line is evaluated.
Tests are usually sold as kits, providing all the items
necessary for performance of the test, such as a dropper for
taking a fixed amount of sample. However, when an
untrained user executes the test, accurate use of the dropper
is not an easy task, resulting in test errors. Application of a
better measuring device for sample addition will improve
this part of the execution of the test. An option is to use an
automatic dispenser; however, when the point of care/need
is restricted to "in the field" this will not be easy, but for
applications where the point of care/need is located in, e.g.,
a feed mill, in the physician's examination room or in the
emergency ward of hospitals, sample addition with an
automated sampling device will increase the accuracy and
reproducibility of the test. It will depend on the purpose of
the test whether this is necessary, taking into account that
the investment costs will increase.
Inherent to the test format, the mounting device has a
restricted volume, often limited to 100 μL; larger volumes,
especially over 1 mL, are precluded. Addition of several
loads of sample will deteriorate the immunostrip, and will
increase the assay time. The development and application
of more sensitive tags is highly recommended here. To
accommodate larger volumes a different assay format
should be used, most probably not a one-step format.
Several attempts have been made to increase the signal
by introducing an enzyme as the label [36, 84, 85] or
electrochemical signal generation [86–88]. Silver enhance-
ment of the response of the gold nanoparticles has been
mentioned as well for sensitivity increase [89–91]. Apart
from losing the "one-step" concept when using an enzyme,
a biological element with limited stability is introduced;
shelf life may decrease and handling becomes more
complicated. The application of a silver enhancer also
requires a second step. When an electrochemical transducer
is used, investment costs might rise to undesirable levels.
Approaches using other physical properties of the label
include the localised surface plasmon resonance of the gold
nanoparticles to increase the signal [66] and surface-
enhanced Raman scattering of gold or silver nanoparticles
[45]. More sensitive chemiluminescent [50] or fluorescent
[24, 47, 48, 92] labels may increase the sensitivity, but will
increase the cost of the assay, because more sophisticated
hardware and software are needed to read the signal. A
typical approach that is used for signal enhancement and
quantitation employs hexacyanoferrate(II)-loaded lipo-
somes in combination with a melittin-labelled analyte
analogue [80]. On the membrane, antianalyte antibody is
immobilised in an antibody competition zone and hexacya-
noferrate(II)-loaded liposomes are immobilised in a signal
generation zone. At the capture zone, analyte and labelled
analyte analogue compete for binding sites of the immobi-
lised antibody. The amount of the melittin conjugate bound
at the antibody site is inversely proportional to the
concentration of the analyte in the sample. The unbound
conjugate can migrate to the signal-generating zone where
the immobilised loaded liposomes are lysed owing to the
action of the melittin, releasing the hexacyanoferrate(II).
The amount of the latter compound can be measured
amperometrically and is directly proportional to the amount
of analyte in the sample.
Raising a sensitive antibody with good selectivity is not
always possible owing to the nature of the analyte under
consideration. Selection of the antibody with the required
properties is dependent on several factors that may not be
compatible with one another. Furthermore, covalent attach-
ment can decrease the affinity for the antigen. Often a
compromise is the only option and consequently the
performance of the test is restricted in one aspect or
another. Selection of a good nucleotide sequence that
enables rapid hybridisation in the NALF format can be
very time-consuming and is often trial and error. Several
authors reported on batchwise implementation of assays
(multiple sample testing), in the microarray format [24, 39,
43] or simply by sticking several strips on a plastic backing
[85]. There are also reports on combined testing of one
sample on several analytes (multianalyte testing) [33, 34,
36, 73, 93]. However, individual tests performed sequen-
tially in high-throughput mode in an automated set-up,
necessary for, e.g., automated progesterone tests in the
milking parlour, are not mentioned.
Inherent to a rapid test, it is advantageous when sample
pretreatment is not necessary. For applications using blood/
plasma/serum or extracts, special sample pad materials are
available that filter cells and/or debris from obstructing the
immunostrip. However, the sample has to be in the liquid
state, and sample pretreatment is obligatory when solid
samples, or dust particles in the atmosphere are analysed.
Even when the sample is liquid, sample cleanup may be
necessary. Also, when the analyte has a very low
concentration, a preliminary concentration step is obligato-
ry. In the nucleic acid lateral flow(immuno)assay set-up, an
amplification step (nucleic acid based amplification, PCR)
is often necessary. Often such protocols can be automated,
but handling will be more complicated and assay time will
578G. Posthuma-Trumpie et al.
Page 11
increase. Still, LFAs can offer rapid results compared with
traditional techniques [39, 41].
Opportunities are mainly related to off-laboratory applica-
tions. The success of the pregnancy test is the best-known
example. Many tests are available nowadays for home
testing; these are non-prescription tests that can be obtained
at the pharmacy or through the Internet, however there is
still an enormous potential left. Examples include testing in
the consulting room of the general practitioner or in the
emergency ward at the hospital, testing on drugs of abuse
for law enforcement, absence of pathogenic organisms in
food/feed, seed and seedlings, forensics and tests related to
biowarfare, where fast results are of utmost importance.
The matrix of these tests is usually blood/serum/plasma
or urine, (extracts of) the food or feed to be tested or
extracts of soil or water for environmental issues. In
biomedical applications the analyte under consideration is
often also present in tears, saliva or sweat, and the use of
these matrices is still under developed, mainly because of
sensitivity aspects. Application of more sensitive tags might
be an option here. Although the restriction on the sample
volume is a weakness (see earlier), it is also an opportunity,
especially when only a limited amount of sample can be
obtained, provided the target analyte is present in sufficient
quantity to meet the sensitivity of the test.
Better reproducibility can be achieved by automation of
the manufacturing process, automated addition of the
sample and controlled drying time and read-out of the
result. Preparation of the strips in a highly reproducible way
using a controlled atmosphere (temperature, humidity, dust)
is a recommendation. One approach is implemented by, e.
g., Biodot, and is described in [94]. The use of an internal
standard for higher reproducibility was mentioned [47, 48].
The lack of published applications in the field of
environmental issues and plant pathology related items is
remarkable. A few applications on the detection of plant
viruses could be found [8, 18, 78]. However, many
commercial kits are available nowadays.. For ecotoxico-
logical items only one publication could be found [58],
apart from the screening of some pesticides in food or feed
(Table 3). LFAs can be of substantial interest in these fields.
Implementation of the microarray format as described in
[39, 43] using an LFA and automated sample addition and
read-out of the results might be an option for some
Compared with biosensor technology (see later) the LFA
technology can be brought to the market extremely quickly
with a relatively small investment. At the time of writing,
the investments in hardware and software for the consumer
are also nil to minimal.
Nowadays the price of hardware and software of many test
systems tends to decrease. This is, among others, due to
miniaturisation of the hardware and reuse of software
components. For some applications in, e.g., the emergency
ward of hospitals, the physician's office, the milking
parlour in the dairy industry, but not in the field, hardware
that was out of the scope a few years ago might now
provide an option for reliable, fast and low-cost tests.
Although gas chromatography-mass spectrometry and
liquid chromatography-mass spectrometry require trained
personnel, applications use far more automation than a few
years ago. Equipment miniaturised and integrated in the
lab-on-a-chip format is already on the market (http://www.
Traditionally enzyme immunoassays are performed in
centralised laboratories by manual intervention of trained
technicians. Nowadays these tests can also be performed
using highly sophisticated hardware in automated set-ups
(ELISA robots). Two research groups designed an auto-
mated enzyme immunoassay set-up for off-laboratory tests
on progesterone in bovine milk in the milking parlour
for information on the reproductive status of dairy cows
[95, 96].
The apoenzyme reactivation immunological system
(ARIS) was rather popular at the end of the twentieth
century [97]. This technique combines recognition of the
analyte by its immunoglobulin (IgG) with signal enhance-
ment by enzymatic processing of the label. Analyte-labelled
flavin adenine dinucleotide can combine with apoglucose
oxidase or can bind to analyte-specific IgG. When analyte
is present in the sample, this will bind to the IgG and the
analyte-labelled flavin adenine dinucleotide will reactivate
the apoprotein to active holoenzyme. Addition of the
substrate glucose generates H2O2, and horseradish peroxi-
dase combined with a chromogen gives the signal that is
linearly related to the amount of analyte in the sample.
Many applications of the ARIS technique are already
available in dry reagent format. These use an Ames
Seralizer reflectometer that was specifically developed for
this assay format [98–101]. Tests are provided for blood/
serum/plasma or saliva. However, recent applications, apart
from the quantitation of trinitrotoluene in drinking water
[102], were not mentioned.
According to the International Union of Pure and
Applied Chemistry, a biosensor is "a self-contained
integrated device, which is capable of providing specific
quantitative or semi-quantitative analytical information
using a biological recognition element (biochemical recep-
tor) which is retained in direct spatial contact with a
transduction element" [103]. The receptor acts upon a
biochemical mechanism, while the transducer can be
Lateral flow (immuno)assay: its strengths, weaknesses, opportunities and threats. A literature survey579
Page 12
diverse in nature, e.g. a chemically modified electrode of
electronic conducting, semiconducting or ionic conducting
material, plasmon-generating, magnetoresistive or interfer-
ometer set-up, and is in direct contact with the biochemical
receptor component. Some immunosensor formats are
surface plasmon resonance, grating coupler, total internal
reflectance fluorometry, magnetobiosensor and interferom-
eter sensor. However, in some publications an LFA with
optical transduction is called a "biosensor" [24, 26, 58, 79].
The magnetobiosensor, under development at Philips
Research, may be a serious threat upon market introduction,
because of its concept of full integration and disposable
assay cartridges [104, 105]. However, from a cost calcula-
tion point of view, applications are primarily foreseen in the
mid- and high-end markets. Introduction of the Young
interferometer sensor as developed at the MESA+Institute
at Twente University, The Netherlands, may be a serious
threat for some applications [106].
Reduction of hardware costs for the microparticle
immunoassay might give this technique a realistic option
in resource-poor settings [107]. This can be an advantage
for multianalyte testing. With use of, e.g., the Luminex®
100 technology, up to 100 different analytes can be tested
in one run in high-throughput mode. At the time of writing,
the price of the microparticles is a hurdle that has to be
overcome. Other brands are available as well, sometimes
for reduced costs. Especially when automated sample
preparation is used this will reduce the assistance of trained
personnel [108]. However, resource-poor settings often are
devoid of electricity, which would hamper the use of such
A format that gets a lot of attention these days is the lab-
on-a-chip format [109–112]. Apart from a much longer
development time, the investments are considerable, as the
whole microfluidic device has to be designed in advance
and prepared using a cleanroom. In addition, the very small
volumes that can be added to the microfluidic device also
implicates that sufficient analyte should be present in such a
small volume to be detectable.
With respect to the patent situation concerning lateral
flow and lateral flow devices, the situation is very
complicated. Several hundred patents have been filed and
Inverness Medical Innovations has collected a number of
important patents on lateral flow over the few last years by
buying strategic companies. However, one of the most
restricting patents on lateral flow is expiring, which would
enable easier market access.
Future perspectives
For commercial applications it is important to improve on
the reproducibility of the tests by automation of sample
addition, recording and data processing.
The use of new labels such as quantum dots and the
upconverting phosphor technology can improve the sensitiv-
ity, allowing for more dilution of the sample with less matrix
interference or use of matrices with lower concentrations,
such as tears or saliva [9, 113]. Multiplexing techniques,
such as labelling the immunoreagents with quantum dots
[37], or upconverting phosphors with different optical
properties is also an option for multianalyte testing [114,
115]. However, the "one-step" principle is lost in an
application as described in [115] as several consecutive
flows were applied. For some multianalyte applications the
recognition elements were reported to be peptides [116] or a
dedicated DNA sequence with hexapet tags [117]. Peptides
can often be synthesised without difficulty, and can sidestep
raising of specific antibodies. However, thorough investiga-
tion of sensitivity and selectivity will be necessary. Nucleic
acid sequences of hexameric repeat tags (hexapet) as capture
probes were reported for detection, and with use of this
approach, discrimination between single nucleotide poly-
morphism sequences [117] was demonstrated.
Applications in the medical field in the Western world
can take advantage of the development of integrating the
LFA in a lab-on-a-chip design [113, 115]. Moreover, often a
clinical laboratory is nearby providing additional facilities.
However, costs will increase substantially. For real off-
laboratory applications at point of care/need in resource-
poor settings, the best option is to use LFAs. Strips can be
stored at ambient temperature for a prolonged time.
However, good recognition elements must be available
and visual qualitative on/off or semiquantitative results
using, e.g., the bar code layout or a reflectometer, must be
supported the study; grant number GPG 06038.
The Dutch Technology Foundation (STW)
Open Access
Creative Commons Attribution Noncommercial License which per-
mits any noncommercial use, distribution, and reproduction in any
medium, provided the original author(s) and source are credited.
This article is distributed under the terms of the
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Tech Consumer Advice
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3D vision with poor eyesight
Likes # 0
Posted February 20, 2012 at 8:07AM
Has anyone any experience of using 3D viewing glasses in conjunction with corrective glasses?
I have found only one manufacturer that mentions their 3D glasses can be used WITH ordinary glasses, and then only in vague terms. As I wear varifocals, and the presciption is different for each eye, I do wonder if I will even be able to use 3D.
Likes # 0
Posted February 20, 2012 at 9:08AM
I have no personal experience but I do not see why 3D glasses cannot be worn over spectacles. One type merely acts as a couple of shutters which open and close in synchronism with the display so that each eye sees the appropriate image. Many stores will have systems on display for one to try so perhaps you could conduct your own test. If this is inconvenient due to your location, I am sure someone on the forum will have a try for you.
Likes # 0
Posted February 20, 2012 at 9:30AM
I had same issue and opted for contact lenses for watching 3D. Trying to ware the 3D glasses over normal one just isn't comfortable or practicably
Likes # 0
Posted February 20, 2012 at 10:43AM
Hi, both my wife and myself wear glasses and we have not found any problem wearing the 3D glasses over them. This is on a Sony system. I'm sure any local retailer would be happy to let you try them.
Likes # 0
Posted February 20, 2012 at 1:55PM
I use the Nividia 3D glasses. No problem with them at all when I wear varifocals or reading glasses
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Miyeon Kwon (MFA 2000) of Prak-Sis Contemporary Art Association is organizing the Axis 2013 International Art Festival: Royal&Loyal, which will take place May 2nd – May 5th at the Ravenswood Event Center in Chicago.
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Salt Lake has been purchased by Betty Mabee and her son, Larry Mabee, to stand at their Golden Eagle Farm near Ramona, Calif., for the 2006 breeding season. A son of the late Deputy Minister, Salt Lake previously stood at Walmac Farm near Lexington.As a sire, Salt Lake is represented by 62 stakes winners, including Champagne Stakes (gr. I) winner Ordway and Salt Champ, winner of the Santa Monica Handicap (gr. I). His progeny, 10 crops of racing age, have earned $37,529,566.Racing for the Young family's Overbrook Farm and trainer D. Wayne Lukas, Salt Lake ended his career with a record of 7-3-4 from 21 career starts. He earned $536,218. His major wins came in the Hopeful Stakes (gr. I), Cowdin (gr. II), Tremont (gr. III), King's Bishop (gr. II), and the Fall Highweight (gr. II).Crook Investment Co. bred Salt Lake in Kentucky from the Queen City Lad mare Take Lady Anne.
Salt Lake to Golden Eagle
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Public protector Thuli Madonsela will get the financial support she needs to expand her staff capacity, while a new central government tender database will soon be launched. These are just two of the key governance-related announcements made by Finance Minister Nhlanhla Nene in his maiden budget in Parliament on Wednesday.
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You Never Give Me Your Money: The Beatles After the Breakup by Peter Doggett
Jane Austen (Canterbury Classics) by Jane Austen
Don Quixote: Complete and Unabridged (Signet Classics) by Miguel de Cervantes Saavedra
Whoever Fights Monsters: My Twenty Years Tracking Serial Killers for the FBI by Robert K. Ressler
Helter Skelter: The True Story of the Manson Murders by Vincent Bugliosi
Marilyn : Norma Jean
Damaged Goods (Pinnacle True Crime) by Jim Henderson
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Gather with birders at the Fish Tale Brewpub on the third Monday of each month. People will share their "best birding spots in our area."
Come ready to share where and why you go birding in a particular spot. People new to our area will benefit from your knowledge. We will gather at the couches in the back.
We just grab a coffee and speak French. Some people have been coming every week for months... it creates a kind of warmth to the group.
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Various machinery/equiment up for sale, see below details including pricing: - One stainless steel, one high tensile and one aluminium mig-welding wires. All 0.90mm in size. Price = $400 for all 3. - 200 litre drum oil pump. Price = $70. - Tanaka chainsaw = $150. - Homelite chainsaw = $120. ...- Baumr chainsaw = $220. - Mulcher = $1050 plus. If interested please contact Steve (owner) on 0. *pick up only* show more
Scania v8 auto opti cruise 6 x 4 prime mover 2003 in very good condition registered super singles and near new all round tyres 90000 GCM air condition power windows immaculate inside air suspension and double chassis
have 390 cu m per hour rated, electric, 3 phase, submersable pump mounted on a flotation frame.frame is 100x150 galvanised box section with two heavy duty plastic flotation pods made by dynaplas.pump is made by EIM electric co. and yes its rated for 390,000 lt per hour. unit is in as new con would s...uit mining application or marine,would be good for cleaning out dams or flooded ship,s holds de watering applications for more info and picturec call or text me on 0 thanks mike show more
4WD Tractor, New Holland, Boomer 25, 3 cylinder diesel engine, 27 HP, 2 Range Hydrostatic Transmission, Single Pedal, Diff Lock, 60 inch Cutting Deck. Excellent condition. Well maintained and stored under cover. Very low hours. Service History and owner manuals available.
If you have got heavy Vehicles or commercial cars and willing to sell it for a top price, please do not hesitate to contact me for a quote. Heavy Vehicles such as: Hino, Volvo, Scania, Isuzu, Mazda, Mercedes and Nissan Commercial Cars such as: Hilux, Vans, Holden Rodeo, Corolla and etc.. ... call Haji on: 0 show more
Solid Cast Brass "EUREKA" 1 I/4" BSP Foot -Valve ......... in OK used condition , Postage $15.00 .
Isuzu FRR 500 2003 model 171 000 km's, beaver tail. Huski 5sdk8 2010 model 1400 hours,tilting hitch,auger unit with augers in various sizes, pallet forks,chain digger attachment,spreader bar. Truck and bobcat both well maintained good tyers all round one owner
FULLY MOBILE ROCK CRUSHING PLANT FOR SALE Primary Crusher Trailer Jaw Crusher – Kang Wong 42"x30" Model FS 4230 Feed Bin into Jaw – track feed type ... Bin – full removal down to top of track feed Secondary Cone Crusher Kang Wong CEC900 3 ft - this is a copy of a Symons. ... show more
Wanted Acetylene or/and Oxy Bottle Full or Near Full Mob 0
mobile crane,300 litre fuel tank,mobile air compressor,fire fighting water tank w/diesel pump,550 kevreck crane,triaxle tilt/slide car trailer,2008 extra cab ford ranger. sale will be held in toodyay on the 5th april 2015.
2012 mazda bt50,stat writeoff,home made san blasting pot,drum cradle,850x250 atlas air compressor,ford ranger spares,oil tanks,ancient gaz producer. sale 5th april 2015 at toodyay
4x unused crusher plates,drill rig hydro control,2x rolls 16 mm unused steel cable,heavy duty draw bar,hydro oil syntectic and standart,spliter box,mercedes benz fully recond truck engine,mercedes benz spares,volvo l50c,4800 hours,forks,4x1 bucket,stick rake,loading scales. sale 5th april 2015 tood...yay. show more
heavy duty hose reel,drill pipe hydro clamp,5 tonne hydro winches,oil coolers,massey 14 disk ,MTU V8 only 55 hours use,ranger px extra cab tub,75 and90mm blue line poly,Edsondrill heads parts. sale 5th of april 2015 in toodyay
24 volt pallet truck,heavy duty 480 volt arc welder,FL6/18 volvo truck w/palfinger crane,triplex pump w/ diesel,workover drill frame,large mono pump w/hydro motor,Cat air filter housings unused,old winch, 2x scania motors,unused hydro hose winch. clearing sale to be held at toodyay on the 5th april... 2015 show more
hydaulic pumps and motors,some recnd,some unused,single phase oil transfer pump,unused welders pipe clamp,Cat C9 unused parts,fuel pump,water pump,ecu,exaust manifold,Cat backhoe unused spare parts,unused socks and wipchecks,unused drilling subs,2012 band saw,275 single phase mig welder. clearing ...sale will be held in toodyay on the 5th april 2015 show more
44hp, turbo, 307 hours, comes with loader and forks, save thousands off new price. Always kept in shed.
We are after old unwanted unused trucks, they can be running or not running, defected, damaged rusty or just used and abused.we need these truck makes: Toyota, Mazda, UD nissan, hino, Mitsubishi, Daihatsu basically any jap truck brand.We can come to you 24/7 anywhere in SA, country SA is a specialit...y, we will pay cash on the spot and organise all transport. Feel free to call text or email us or send a pick and we will be back to you promptly.even truck drive trains etc. show more
20 kva generator,perkins 3 cylinder,single phase bench grinder,sockets and plugs single and three phase,old english pedestal drill,elliot brand good working condition,air compressor air dryer,6 unused trailer air bags,6 unused trailer axle drums,lots of hydraulic fittings unused,7.5 hp single phase ...motor. sale will be held in toodyay on the 5th april 2015 show more
Grey Fergy. With Original service Mobilubrication chart. Fully hardened respray. New battery, steering box seals, gearbox input shaft seal, distributor cap and leads. Rewired. Near new tires with front wheel weights. 4.0-19, 11.2-28. Roll bar with canopy. New engine oil, filter and transmission oil.... Tuned governor. Brakes serviced. Runs very well. $2500.00 PH 0. show more
EPW,10 metre,electric hydraulic,in good nick,.hydro power pack,18hppetrol motor,as new.high pressure cleaner,13hp motor,little use.unused drilling head w/hydro motor,heavy duty.mono pump w/hydro motor large.mono pump w/hydro motor. clearing sale to be held in toodyay,on the 5th april2015.
WOWW!!!!!!!! Finally a Good Quality Diesel Powered UTV Side by Side with all round durability that won't break the budget. This brilliant piece of machinery comes standard with :- 1. A reliability Perkins 403D-07 3 Cylinder Diesel engine with CVTECH Transmission, 2. Brilliant 4WD Capa...bilities, 3. Increased Load Capacity (300Kgs) 4. Approved Roll Over Protection, 5. Steel Lined Tilt Tray, 6. Heavy Duty Steel Wheels. 7. Heavy Duty 6 Ply R Tyres ... show more
These are one of the most versatile machines you can get to compliment your workforce. With independant heavy duty suspension, lockable 4WD and a guttsly 600cc EFI engine, this machines performs like no other arouind the farm or worksite. Ready for conditional rego they are fully equiped with many s...tandard accessories including tow bar, winch, full lighting and actuated tipper, 4 wheel disc brakes and even an MP3 player. NOW WITH A FULL 12 MONTHS FACTORY WARRENTY (Conditions Apply) EZI ... show more
TRANSPORTING TRACTORS, TRUCKS, MACHINERY, ETC PLEASE LEAVE A NUMBER IF REQ A QUOTE ( NO PRIVATE NUMBERS ) Can carry 4.5 tons x 28' tray ring for quote, as i dont check emails that much will travel almost anywhere
A Rolabar hay rake, in good condition, all tynes there, working order. Sale due to property sold. See also seeder. Inspection welcoem phone david 0
Horse drawn drill converted to use by tractor, in excellent working order, gears to change sowing rates also included, Seed and fertiler chart (photocopy) available. The pictures tell the story, shedded with bags covering to protect against the moisture. Fill with seed and super and start sowin...g - no repairs required. First to see will buy. See also rake Inspection welcome phone david 0 show more
Caterpillar D2 Dozer, PTO with 3 point linkage to attach Auxiliary implements. 24v Electric starter, always starts first time. Caterpillar Diesel runs like brand new. Two Brand new lifting rams. Brand new cutting edge supplied. PRICE IS NEGOTIABLE, Come and have a look and make an offer. You wont be... dissapointed. show more
ISUZU 85 NKR TRAY TRUCK 2100 x 3000 drop side tray recond gear box new clutch and bearings diff reshim brakes rebuild new injectors new slave cylindors new windscreen/wipers and motor new lights new bumber bar new heater new headlining spair bucket seat drives great was project bu...t no longer doing due to illness Bargain $ 3500.00 ONO show more


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NEW YORK (CNNMoney.com) -- A congressional committee is widening its investigation of a drug making arm of Johnson & Johnson to include an attempted 2009 "phantom" recall of Motrin.
The House Committee on Oversight and Government Reform was already looking into several recalls by the division, McNeil PPC, and charges by the Food and Drug Administration of sloppy drug-making processes.
Now it is looking into FDA charges that in June 2009, McNeil attempted to disguise its efforts to remove Motrin from store shelves without initiating a formal recall.
After the FDA learned of the effort, it prompted McNeil to initiate a nationwide recall of more than 88,000 tablets of Motrin because of a problem with the way the drug dissolved, according to an FDA enforcement report.
The FDA said it became aware of the alleged Motrin scheme after it obtained a memo that detailed instructions to contract workers to buy up Motrin from stores. The memo was first sent anonymously to Oregon state regulators.
"You should simply 'act' like a regular customer while making these purchases. There must be no mention of this being a recall of the product!" reads the memo dated June 12, 2009.
The FDA could not identify the other companies involved, named in the memo as WIS and CSCS.
The FDA said it was aware that McNeil was using a contractor to remove a sample batch of the product, but not at such a wide scale.
"Once we learned of the contractor's activities, the FDA asked McNeil to initiate a recall, and the company complied," the agency said in a statement.
The FDA subsequently classified the Motrin recall as Class 2, meaning the recalled products have a low chance of causing major injuries or death, but still pose some risk of "adverse events."
It appears that McNeil never issued a press release about the recall. Companies are not required to issue press releases for Class 2 recalls, though most companies do, as did McNeil for four other Class 2 recalls in the past year.
The FDA, which typically posts such press releases, confirmed that it does not have a press release from McNeil about the Motrin recall.
McNeil would not comment on whether it publicly disclosed the recall. The House committee has given Johnson & Johnson until 4 p.m. on June 7 to provide more information and records. McNeil said it is looking into the memo and "will respond back to the [congressional] committee."
FDA spokeswoman Elaine Gansz Bobo said the agency could not yet determine if McNeil violated any regulations with its actions.
"This product appears to have been adulterated because it didn't meet dissolution specifications," said David Rosen, who worked at the FDA for 14 years and now advises major pharmaceutical companies on FDA regulation and compliance. However, Rosen said the law prohibits a company to introduce or deliver a product that is adulterated or misbranded into the marketplace.
McNeil Consumer Healthcare, a division of Johnson & Johnson, has initiated four recalls of its over-the-counter drug products such as Tylenol, Motrin, Zyrtec in the past eight months.
The drugmaker last month shut production at its Fort Washington, Pa., plant where its recalled children's drugs were manufactured and pledged to remedy serious quality and safety lapses at its manufacturing facilities.
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Guinness owner says inclusion of an LGBT group in this year's parade is reason to resume its sponsorship, though some gay rights groups are still protesting the parade's policies. More |
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The Stratos Card is a new Bluetooth-connected device that wants to make all of your credit cards, debit cards, loyalty cards, and gift cards obsolete. More |
Self-described 'Last of the Mohicans' is still fixing typewriters after 55 years. More |
Last week, the House of Representatives passed a bill that would expand benefits offered by the college savings plans to include coverage of expenses like computers and Internet access, among other things. And there is bipartisan support for it in the Senate, too. More |


text:
James Pond 2: Codename: RoboCod
Published by Developed by Released Platforms |
The title music by Richard Joseph bares much similarity to the theme from RoboCop (the movie). Some notes are changed, and the music is obviously much "cuter", but you can still hear the RoboCop theme in there!Product placement
The UK release of the game featured considerable product placement from Penguin chocolate biscuits, making it one of the first games to use this form of marketing (Zool is another famous example from this era).Awards
- Commodore Format
- July 1993 (Issue 34) - Modern Classics: Platformers (Check 'em out)
- Retro Gamer
- September 2004 (Issue #8) – #67 Best Game Of All Time (Readers' Vote)
- ST Format
- January 1993 (issue #42) - #17 in '50 finest Atari ST games of all time' list
Contributed by Martin Smith (63052) on Mar 07, 2005. [revised by : *C64* (54133), Patrick Bregger (119826) and Jo ST (16310)]. -- edit trivia