The diagnosis of tuberculous meningitis using the polymerase chain reaction.
OBJECTIVE
DNA amplification by the polymerase chain reaction (PCR) was evaluated as a means for rapid diagnosis of tuberculous meningitis (TBM).


METHODS
A 240 bp region (nts 460-700) from the MPB 64 protein coding gene specific for Mycobacterium tuberculosis (TB) was selected for amplification. Nineteen clinical samples were studied. Six were obtained from patients with TBM diagnosed by culture (4/6) or by response to therapy (2/6). The remaining 13 samples were obtained from patients with febrile seizu es (8/13), aseptic meningitis (3/13) and septic meningitis (2/13), and these served as negative controls.


RESULTS
We detected TB DNA in all the 6 CSF specimens obtained from patients with TBM. PCR alone was sufficient to detect TB DNA in 5 of these 6 samples. However, one sample was positive only when PCR was followed by oligonucleotide hybridisation. In the 2 patients whose CSF were obtained only after commencement of TB therapy, TB cultures were negative but positive on PCR nd oligoprobe labelling. The diagnosis of TBM was confirmed based on their remarkable response to therapy. Twelve of the thirteen negative controls were TB DNA negative. There was one false positive sample, which was thought to be due to TB DNA contamination.


CONCLUSIONS
Taken together, our results indicate that DNA amplification using PCR, followed by oligonucleotide hybridisation offers a rapid (5 working days) means of diagnosis of TBM, provided care is taken to ensure that cross contamination of DNA samples is avoided.
Lee
B W
BW
Department of Paediatrics, National University Hospital, Singapore.
Tan
J A
JA
Wong
S C
SC
Tan
C B
CB
Yap
H K
HK
Low
P S
PS
Chia
J N
JN
Tay
J S
JS
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D005784:Gene Amplification; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D008582:Meningitis, Aseptic / Q000175:diagnosis / Q000382:microbiology; D016920:Meningitis, Bacterial / Q000175:diagnosis / Q000382:microbiology; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D003294:Seizures, Febrile / Q000175:diagnosis / Q000382:microbiology; D014390:Tuberculosis, Meningeal / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000382:microbiology
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D009841:Oligonucleotides