Lee B W, Tan J A, Wong S C, Tan C B, Yap H K, Low P S, Chia J N, Tay J S
Department of Paediatrics, National University of Singapore.
J Neurol Sci. 1994 May;123(1-2):173-9. doi: 10.1016/0022-510x(94)90220-8.
DNA amplification of three Mycobacterium tuberculosis-specific DNA sequences by the polymerase chain reaction (PCR) were evaluated as a means for rapid diagnosis of tuberculous meningitis (TBM). The DNA sequences amplified were a 123 bp region of the IS6110 insertion elements which occur in multiple copies in the mycobacterial genome, a 240 bp region (nts 460-700) from the MPB 64 protein coding gene, and the 383 bp region of the 65 kDa heat shock protein (HSP) antigen. Twenty-seven cerebrospinal fluid (CSF) specimens were studied. Six were obtained from patients with TBM diagnosed by culture (4/6) or by the patients' response to anti-tuberculous therapy (2/6). The remaining 21 specimens were obtained from patients with febrile seizures (3/21), aseptic meningitis (3/21), septic meningitis (14/21), and cryptococcal meningitis (1/21), and these served as negative controls. Our results indicate that although the protocols involving the 3 DNA sequences were able to detect TB DNA in the 6 TBM specimens, the main drawback was their extreme sensitivity, thus giving rise to false positive results. In particular, the repeat copy sequence, IS6110, and the 65 kDa HSP gave unacceptably large numbers of false positive results (62% and 33%, respectively).
通过聚合酶链反应（PCR）对三种结核分枝杆菌特异性DNA序列进行DNA扩增，以此作为快速诊断结核性脑膜炎（TBM）的一种方法进行了评估。扩增的DNA序列包括：在分枝杆菌基因组中以多拷贝形式存在的IS6110插入元件的一个123 bp区域、来自MPB 64蛋白编码基因的一个240 bp区域（核苷酸460 - 700）以及65 kDa热休克蛋白（HSP）抗原的383 bp区域。对27份脑脊液（CSF）标本进行了研究。其中6份取自经培养确诊为TBM的患者（4/6）或对抗结核治疗有反应的患者（2/6）。其余21份标本取自热性惊厥患者（3/21）、无菌性脑膜炎患者（3/21）、化脓性脑膜炎患者（14/21）和隐球菌性脑膜炎患者（1/21），这些作为阴性对照。我们的结果表明，尽管涉及这3种DNA序列的方案能够在6份TBM标本中检测到结核DNA，但其主要缺点是灵敏度极高，从而导致假阳性结果。特别是，重复拷贝序列IS6110和65 kDa HSP产生的假阳性结果数量多得令人无法接受（分别为62%和33%）。
