PMC3656678_01
a-c Spinal cord MRI findings. a Sagittal T2-weighted MRI (TR 3809, TE110) of the cervical spine (c) in patient 1. A high-signal-intensity lesion at the cervical spinal nerve 2 is shown. b Sagittal T2-weighted MRI (TR 3797, TE 110) of thoracic spine in patient 2. c Axial T2-weighted MRI (TR 539, TE 18) of thoracic spine in patient 2. A high-signal-intensity lesion at Th 3-4 is shown. d, e Nested PCR assay results for the CSF specimen collected on admission. d Patient 1, e patient 2. * Positive result. Lane 1: molecular weight marker, lane 2: internal control (human beta-globin: 196 bp), lane 3: negative control (clinically isolated Staphylococcus aureus DNA), lane 4: case, lane 5: positive control (172 bp), lane 6: negative control, lane 7: case, lane 8: positive control (88 bp), lanes 3-5: single-PCR assay results, lanes 6-8: nested PCR results. As a positive control, the original plasmid into which a DNA fragment (172 bp) of the ORF-29 gene had been inserted was prepared. The nucleotide sequence of the inserted DNA fragment was confirmed by direct sequencing in both strands. The extracted DNA specimens from clinically isolated Staphylococcus aureus were used as a negative control. In addition, as an internal control, a pair of primers specific to the human beta-globin gene (GenBank accession no. L48217) was prepared. The 196-bp human beta-globin gene fragment as an internal control was amplified in another tube under the same assay conditions. In conventional single-PCR assay, a weak 172-bp band was only shown at lane 4 for patient 1. In contrast, the nested PCR assay demonstrated clearly positive 88-bp bands at lane 7 for both patients.
crn-0005-0081-g01.jpg
PMC3656678_02_crn-0005-0081-g01.jpg
F1
Spinal cord MRI showed a high-signal-intensity lesion at Th 3-4 on a T2-weighted image (fig. 1b and c). IgG antibody index was elevated against VZV (8.24), HSV-1 (7.30) and CMV (5.15), which suggested intrathecal synthesis of IgG antibodies (table 3). In the CSF specimen collected on admission, no VZV DNA was detected by conventional commercial single-PCR assay (table 3), but our original nested PCR assay revealed a definite positive result (table 3 and fig. 1e).
PMC3656678_02
a-c Spinal cord MRI findings. a Sagittal T2-weighted MRI (TR 3809, TE110) of the cervical spine (c) in patient 1. A high-signal-intensity lesion at the cervical spinal nerve 2 is shown. b Sagittal T2-weighted MRI (TR 3797, TE 110) of thoracic spine in patient 2. c Axial T2-weighted MRI (TR 539, TE 18) of thoracic spine in patient 2. A high-signal-intensity lesion at Th 3-4 is shown. d, e Nested PCR assay results for the CSF specimen collected on admission. d Patient 1, e patient 2. * Positive result. Lane 1: molecular weight marker, lane 2: internal control (human beta-globin: 196 bp), lane 3: negative control (clinically isolated Staphylococcus aureus DNA), lane 4: case, lane 5: positive control (172 bp), lane 6: negative control, lane 7: case, lane 8: positive control (88 bp), lanes 3-5: single-PCR assay results, lanes 6-8: nested PCR results. As a positive control, the original plasmid into which a DNA fragment (172 bp) of the ORF-29 gene had been inserted was prepared. The nucleotide sequence of the inserted DNA fragment was confirmed by direct sequencing in both strands. The extracted DNA specimens from clinically isolated Staphylococcus aureus were used as a negative control. In addition, as an internal control, a pair of primers specific to the human beta-globin gene (GenBank accession no. L48217) was prepared. The 196-bp human beta-globin gene fragment as an internal control was amplified in another tube under the same assay conditions. In conventional single-PCR assay, a weak 172-bp band was only shown at lane 4 for patient 1. In contrast, the nested PCR assay demonstrated clearly positive 88-bp bands at lane 7 for both patients.
crn-0005-0081-g01.jpg
PMC3656678_02_crn-0005-0081-g01.jpg
F1
Spinal cord MRI showed a high-signal-intensity lesion at Th 3-4 on a T2-weighted image (fig. 1b and c). IgG antibody index was elevated against VZV (8.24), HSV-1 (7.30) and CMV (5.15), which suggested intrathecal synthesis of IgG antibodies (table 3). In the CSF specimen collected on admission, no VZV DNA was detected by conventional commercial single-PCR assay (table 3), but our original nested PCR assay revealed a definite positive result (table 3 and fig. 1e).