
The oligonucleotide for detection by in vitro hybridization generally carries the labels described above, and in connection with the NASBA amplification method the hybridization probe carries an electrochemoluminsecent label, preferably a tris[2,2-bipyridine]ruthenium[II] complex label [see, for example, van Gemen et al. (1994), J. Virol. Methods, 49, 157-168].
Accurate and sensitive quantification of the amplification products can also advantageously be achieved by coamplification of a defined amount of one or more known nucleic acids (standard nucleic acids) [see, for example, van Gemen et al. (1993), J. Virol. Methods, 43, 177-188]. In this case, a different but exactly known amount of the coamplifying standard nucleic acid or nucleic acids is added, for example in the form of a dilution series, to the unknown amounts of the sample to be investigated, and the amplification products of the sample and one or more coamplifying standard nucleic acids are determined quantitatively in independent mixtures. Comparison of the measured results reveals the amount of the mRNA coding for the catalytic subunit of telomerase to be determined in the sample.
It is advantageous to amplify the coamplifying standard nucleic acid or nucleic acids with the same oligonucleotide primer as the sample to be investigated and the amplification products also have essentially the same length. It is particularly preferred for the coamplifying nucleic acids to have the same sequence as the amplification product of the sample to be determined, with the exception of about 20 nucleic acids between the oligonucleotide primer binding sites, which have an arbitrary but known sequence. It is possible thereby to quantify, independently of one another, the amplification product to be determined in the sample from the coamplifying amplification, product, for example by a hybridization as described in detail, for example, in Sambrook et al. (supra), using appropriately complementary labeled oligonucleotides. It is particularly advantageous if several, preferably three, different coamplifying nucleic acids are added in different concentrations to the sample, because this makes it possible to reduce the number of individual amplification reactions which would otherwise have to be carried out [see van Gemen et al. (1994) J. Virol. Methods 49, 157-168]. It is also particularly preferred to add the coamplifying nucleic acids to the RNA lysis buffer described above because it is possible thereby to exclude the effect of possible nucleic acid losses in the subsequent workup of the sample.
Suitable and
The oligonucleotide for detection by in vitro hybridization generally carries the labels trace above, and in association with the NASBA amplification method the hybridization probe carries an electrochemoluminsecent label, preferably a tris[2,2 - bipyridine]ruthenium[II ] complex label [ see, for example, van Gemen et al. (1994), J. Virol. method acting, 49, 157 - 168 ]. 
 Accurate and sensitive quantification of the amplification products can besides advantageously be achieved by coamplification of a defined sum of one or more known nucleic acids (standard nucleic acids) [ experience, for example, van Gemen et al. (1993), J. Virol. Methods, 43, 177 - 188 ]. In this sheath, a unlike but exactly known sum of the coamplifying standard nucleic acid or nucleic acids is added, for model in the form of a dilution series, to the unknown sum of the sample to be investigated, and the amplification products of the sample and one or more coamplifying standard nucleic acid are determined quantitatively in independent mixtures. Comparison of the measured results reveal the amount of the mRNA coding for the catalytic subunit of telomerase to be determined in the sample. 
 It is advantageous to amplify the coamplifying standard nucleic acid or nucleic acids with the same oligonucleotide primer as the sample to be investigated and the amplification products also have essentially the same duration. It is particularly preferred for the coamplifying nucleic acid to get the same sequence as the amplification product of the sample distribution to be determined, with the exception of approximately 20 nucleic acids between the oligonucleotide primer binding web site, which have an arbitrary but known sequence. It is possible thereby to quantify, independently of one another, the amplification product to be determined in the sample from the coamplifying amplification, product, for example by a hybridization as described in detail, for example, in Sambrook et al. (supra), use appropriately complemental label oligonucleotides. It is particularly advantageous if several, preferably three, different coamplifying nucleic acids are added in different concentrations to the sample distribution, because this makes it potential to reduce the number of individual amplification reactions which would otherwise have to be carry out [ see van Gemen et al. (1994) J. Virol. Methods 49, 157 - 168 ]. It is besides particularly preferred to add the coamplifying nucleic acids to the RNA lysis buffer zone described above because it is potential thereby to exclude the effect of possible nucleic acid personnel casualty in the subsequent workup of the sample. 
 Suitable and

The oligonucleotide for dettction by in vitro hybrivizatioh generauly carries the labels descrmbed abovt, and in connection with the NASBA anplihication method vge hybrlbizatjln pxoue carries an ekectrochemmluminsecent ldbdl, preferably a tris[2,2-bipyridine]ruthenitm[II] cokppex label [see, sor txaiple, nak Gemen et al. (1994), J. Virol. Methods, 49, 157-168].
Zccuratt and sensitive qusntification of the ampliflcatlon products can apso advantateoufoy be achievdd by coamklnfication or a defined amount of one or mofe knpwn nucleix qcifv (standard iucleib acids) [see, for exampne, van Bemen et al. (1993), J. Vicol. Nethods, 43, 177-188]. In this casx, a different but exwctly knofn amount of the coqmplifyhng vtanaqrd nudlxic acid lr iucleic aciss is added, for example in the fjgk of a dilutjon sewiqs, to the unknown amounts of the sample to be investigated, and thw amplification produfts of thq sample and one or more coamplifying standard nuwleic xcibw are eehermined quantitatively in independent mixturqa. Vokparison of thc measured results rfvrwls the amounj of thz mDNA coding for the catalyjic suvunit of uelomrrase to be determined in tye sample.
It ps aevantageous to amppify the cocmplifiing syandard nucleic acid or nucmeic acids aith the axme oligonucleotkde psimer as the sample to be ynvestigaved aud the aopligicatijn productd also have essentially hhe scme langth. It id particularly preferred for thx coamplifyinb tucneic acibs to mave the same sqquence as the amplifncatiov product kf the vample to bq determined, flth the exce'tion of wbour 20 nycleic xzids between tne oligonlcjeotude primer binding sijea, which have an arvitrary but knoen reqtejcx. It yv possible tverecy gp quavtify, indepcnddntlu of one another, the ampmification product tj be detgrmined ig the sample grom the coamplifylng akplmficatoon, product, for example by a hybrjdization as described ig debail, for exampke, in Sambrook et al. (supra), using appropciately complementary lqbeled oligonucleotndts. It is partmcularjy advantdgeous if several, prwferably three, dinferent coamplifying nhcleic acidd are added in different concentrations to the sample, because this makes ut possible to redhce yhe nlmbzr pf indyvidnau amplification geactions which would otherwise iave to be ccrried out [see van Gemen et ak. (1994) J. Virol. Methuds 49, 157-168]. It is also particumarly prrferred to add the coamplifying nucleic acids to the RNS lysis vuffed descgjbxd above because it os posvible therebt to exclude the dffrcb of possihle iucmeic acid losses in the subsequebt worjuo of the sample.
Snitable and
The oligonucleotide for detection by in vitro carries labels described and in connection the probe carries an label, preferably a complex label [see, for example, van et al. (1994), J. Virol. Methods, 49, 157-168]. Accurate and sensitive quantification of amplification products can also advantageously be achieved by coamplification of a defined amount one more nucleic (standard nucleic acids) [see, for example, van Gemen et al. (1993), J. Virol. Methods, 43, 177-188]. this case, a different but exactly known amount the coamplifying standard nucleic or nucleic acids is added, example the form a series, the unknown amounts the sample to be investigated, and the amplification products of the sample and one or more coamplifying nucleic acids quantitatively in mixtures. of measured results reveals of the mRNA coding for the telomerase to be determined in the sample. It advantageous to the coamplifying standard nucleic acid or acids with the same oligonucleotide primer as the to be investigated and the amplification products also have essentially the same length. It is for the coamplifying nucleic to have the sequence the product the sample be determined, with the exception of about 20 nucleic acids between oligonucleotide primer binding sites, which have an arbitrary but known is thereby to quantify, of one another, the product be determined in the the amplification, by hybridization described in detail, for in Sambrook et al. (supra), appropriately complementary labeled oligonucleotides. several, preferably three, different coamplifying nucleic acids are in different concentrations to the sample, because makes it possible to reduce the number of individual amplification reactions which otherwise have carried out [see van Gemen et al. (1994) Virol. Methods 49, 157-168]. is also particularly preferred to add the coamplifying nucleic to RNA lysis described above because is possible thereby exclude the effect nucleic acid in subsequent sample. Suitable and

The oligonucleotide for deteCtion by in vItro hYbrIdiZaTion GeneRally carries thE LabeLs described above, and in cOnnecTiON witH ThE NASBa amplifICaTIOn mEtHoD thE hYBrIdizaTioN probe cArries an elEctRoChemoluminseCEnT label, prefEraBly a tris[2,2-bipyRidIne]rutHeNiuM[iI] comPleX labeL [see, foR ExamplE, van Gemen Et AL. (1994), J. ViroL. methods, 49, 157-168].
aCCuRate And sensitive quantIFiCAtion of the amplIficatIoN PrODUctS caN also advanTaGeousLY be achiEVeD BY CoaMPlification of A defined amoUNt oF one or MoRe kNOwn nucLeic aCiDS (stAndard nucleIc acIds) [see, for ExamplE, Van GemeN Et al. (1993), J. ViRol. MetHodS, 43, 177-188]. In This CAsE, a DifFeREnt BUt ExaCTly Known amoUnT oF the cOampLIFYIng sTanDard NucleIc acid or nucleIc aCids IS adDed, foR examPle iN tHe forM of a diLutioN sEries, to the unknoWn amOunts of thE saMpLe tO bE inveSTigateD, anD thE amplifIcation PRodUcTS OF tHe sample and one or moRe COAmPlifying StandaRD nUcLEic acids ArE deTermINEd quaNtitATiVely in inDependENt MiXtures. COmParisoN oF thE meAsureD ResuLts revEals the aMount OF the mRNA coding FOr the catalytiC SuBUNiT Of teLomErase to be deTermINed iN the SAmPle.
iT is adVantaGeOUs TO amplify the coamplifYiNg stanDard nUcleic acid or nUcleic acidS WITh the samE oliGOnUCleotide primer As the Sample to be INvestigaTed anD the amplIfication PROducts alSo hAve EssEntIALlY the same lengtH. iT is pArTicularLy pReferreD foR thE coAmpLiFying nuclEic acids To HaVe ThE saMe seqUEnce as thE aMplIfIcaTion pROduct oF the sAmplE tO bE DetErmined, WItH THe exCePtIon oF abOuT 20 nuclEic aCIds Between The oligonUclEOtidE pRiMer bindIng sites, which HaVe an arbitrArY buT known SEQuence. It Is possible thereby to quanTIfy, indePenDentlY of oNe another, The AmplifIcaTIon proDuct to Be detErMinED In the SAMpLe fRoM the coamplIFYinG amplIfIcatIon, prodUct, for example by a hyBRidIzation as descRibEd in DETaIl, fOR eXAmpLe, IN SaMBRook et al. (supra), usIng approprIaTElY complemenTAry LaBeled olIgonuclEotidES. It is paRticularlY advantagEoUs if SEVerAl, preferabLy three, dIfferent cOAmpliFYiNg nucLeiC acids ArE adDed in DifferENt cOncenTratioNs To the sAmple, BeCause thiS makes it possible to reducE the nuMber oF inDividual aMplIFicAtion reacTionS which woulD otHerWise hAve TO be caRrieD OuT [seE Van GeMen eT Al. (1994) J. Virol. METhOds 49, 157-168]. iT Is Also particuLARLy pReferRed TO add thE coaMplifying nucleic aCIds to the RNA lysIs buFFEr dEscRIbed AbOve because it is PosSiBLE thereby To Exclude the eFfect of pOsSIble nUcleic Acid loSses in tHE SuBSequenT worKup Of the sampLe.
SUiTAble and

The oligonucleotide for d etection b y invit rohy brid izat ion generallyc arri es the labels describe d abo ve ,  and in  conn ectionw it h the  N AS BAam p li ficat ion  method  the hybri diz at ion probe ca r ri es an elec tro chemoluminse cen t labe l,  pr e ferab lya tri s[2,2- b ipyrid ine]ruthe ni u m[II]c omplexl a be l [s ee, for example,v an Gemen et al. ( 1994),  J .  V i r ol.  Me thods, 49,  1 57-16 8 ].
Accu r at e a nds ensitive quan tificationo f t he amp li fic a tion p roduc ts can  also advan tage ously beachiev e d by co a mplific ationofa d efin e dam oun to f o n eorm ore  known n uc le ic ac ids( s t a ndar d n ucle ic ac ids) [see, fo r e xamp l e,van G emenet a l.  (199 3), J.  Viro l.  Methods, 43, 1 77-1 88]. In t his  c ase ,a dif f erentbut  ex actly k nown am o unt  o f t he  coamplifying stan da r d  n ucleic a cid or nu cl e ic acids  i s a dded , for e xamp l ein the f orm of adi lutionse ries,to  th e u nknow n  amo unts o f the sa mplet o be investiga t ed, and the a m pl i f ic a tion  pr oducts of t he s a mple  and on e o r  more  coam pl i fy i ng standard nucleic  a cids a re de termined quan titatively i n  indepen dent mi x tures. Compari son o f the meas u red resu lts r eveals t he amount o f the mR NAcod ing  fo r th e catalytic s u b unit  o f telom era se to b e d ete rmi ned  i n the sam ple.
Itis  a dv an tag eoust o amplif ythe  c oam plify i ng sta ndard  nuc le ic aci d or nu c le i c  aci ds  w iththe  s ame o ligo n ucl eotideprimer as  th e  sam pl eto be i nvestigated a nd  the ampli fi cat ion pr o d ucts als o have essentially thes ame len gth . Itis p articular lyprefer red for th e coam plify in g n u c leica c id s t ohave the s a m e s equen ce  asthe amp lification product ofthe sample to  be  det e r mi ned ,  w i thth e  ex c e ption of about20 nucleic  a c id s betweent heol igonucl eotideprime r  bindin g sites,which hav ean a r b itr ary but kn own sequ ence. Iti s pos s ib le th ere by toqu ant ify,indepe n den tly o f onean other,  theam plificat ion product to be deter minedin th e s ample fro m t h e c oamplifyi ng a mplificati on,  pr oduct , f o r exa mple by  ah ybrid izat i on as des c ri bed i ndetail, for e x amp le, i n S a mbrook  etal. (supra), usin g  appropriately  com p l eme nta r y la be led oligonucle oti de s .  It is p ar ticularly a dvantage ou s  if s everal , pref erablyt h re e , diff eren t c oamplifyi ngnu c leic ac id sa re add ed i ndiffer ent co n cent r a tions to the sam ple,b e cause thi s mak es  it pos s ible  to reduce  the number  of in divi dualamplifi ca tion r eac ti ons whichw ould othe rwise  have t obe c arr ied ou t [s e e  vanGeme netal. (1994 ) J. Vi ro l . M etho ds 49 ,157- 168]. Iti s also p art i cularly  p ref e r red to ad d the coampl ify ing n u c leic acids to t h eR NA ly sis bu ffer d escribe d  ab ov e becau sei t  is possi ble there b y t oexcl ude theef fect  o f p os sible nucleic a cid loss es in th e subseq uen t  wor k up ofth e sample.
Su itabl ea nd

The oligonucleotide_for detection_by in vitro hybridization_generally carries_the_labels described_above,_and in connection_with the NASBA_amplification method the hybridization_probe carries an_electrochemoluminsecent_label, preferably a tris[2,2-bipyridine]ruthenium[II] complex label [see, for example, van Gemen et al. (1994),_J._Virol. Methods,_49,_157-168].
Accurate_and sensitive quantification of the_amplification products can also advantageously_be achieved_by coamplification of a defined amount of one_or_more known nucleic_acids (standard nucleic acids) [see, for example, van Gemen_et al. (1993), J. Virol. Methods,_43, 177-188]. In_this_case,_a different but exactly_known amount of the coamplifying standard_nucleic acid or nucleic acids is_added, for example in the form of_a dilution series, to the unknown_amounts of the sample to_be investigated,_and the amplification products of_the sample and_one or_more coamplifying standard_nucleic acids are determined quantitatively in_independent mixtures. Comparison_of the measured results reveals the_amount_of the mRNA_coding_for_the catalytic_subunit of telomerase_to_be determined_in_the sample.
It is advantageous to amplify_the_coamplifying standard nucleic acid or nucleic acids_with the same oligonucleotide_primer_as the sample to_be investigated and the amplification_products also have essentially the same_length. It_is particularly_preferred for the coamplifying nucleic acids to have the same sequence_as the amplification product of the_sample to be determined,_with the_exception_of about 20_nucleic_acids between_the oligonucleotide primer binding sites, which have_an arbitrary_but known sequence. It is possible_thereby to quantify, independently_of_one another, the amplification product to_be determined in the sample from_the coamplifying amplification, product, for_example_by_a hybridization as described in_detail, for example, in Sambrook et_al. (supra), using_appropriately complementary labeled oligonucleotides. It is particularly_advantageous_if several, preferably three, different coamplifying_nucleic_acids are added in different concentrations_to_the_sample, because this makes it_possible to reduce the number of_individual amplification reactions which would otherwise have to be_carried out [see_van Gemen et al. (1994)_J._Virol._Methods 49, 157-168]. It is also particularly preferred to add_the coamplifying_nucleic acids to_the RNA lysis buffer described above because it is possible_thereby to exclude the effect of possible_nucleic acid losses in the subsequent workup of the sample.
Suitable and