Takahashi Teruyuki, Tamura Masato, Takahashi Sachiko Nonaka, Matsumoto Koichi, Sawada Shigemasa, Yokoyama Eise, Nakayama Tomohiro, Mizutani Tomohiko, Takasu Toshiaki, Nagase Hiroki
Advanced Research Institute for the Sciences and Humanities, Nihon University, and Department of Internal Medicine, Nihon University Nerima-Hikarigaoka Hospital, Tokyo, Japan.
J Neurol Sci. 2007 Apr 15;255(1-2):69-76. doi: 10.1016/j.jns.2007.01.071. Epub 2007 Mar 9.
Although the "gold standard" for diagnosis of tuberculous meningitis (TBM) is bacterial isolation of Mycobacterium tuberculosis (M. Tb), there are still several complex issues. Recently, in the diagnosis of TBM, the detection of M. Tb DNA in cerebrospinal fluid (CSF) samples using PCR has been widely performed as more rapid, sensitive, and specific diagnostic method. Based on Taq Man(R) PCR, the authors developed a novel technique of internally controlled quantitative nested real-time (QNRT) PCR assay that provided a prominent improvement in detection sensitivity and quantification. Total 43 CSF samples from 8 serial patients with suspected TBM were analyzed. The CSF samples were collected before and during standard anti-tuberculosis treatments (ATT). The QNRT-PCR assay revealed positive results for 24 out of 43 serial CSF samples (55.8%) collected during the treatment course of ATT. Moreover, the bacterial cell (BC) numbers of M. Tb analyzed by the QNRT-PCR assay decreased gradually, correlating with the improvements of the patient's clinical conditions. Since the QNRT-PCR assay provides the ability to calculate a numerical value for the initial BC numbers of M. Tb in CSF samples, this method is an extremely useful and advanced technique for use in assessing the clinical course of TBM.
虽然结核性脑膜炎（TBM）诊断的“金标准”是结核分枝杆菌（M. Tb）的细菌分离，但仍存在几个复杂问题。最近，在TBM诊断中，使用聚合酶链反应（PCR）检测脑脊液（CSF）样本中的M. Tb DNA已作为一种更快速、灵敏且特异的诊断方法被广泛应用。基于Taq Man® PCR，作者开发了一种新型的内控定量巢式实时（QNRT）PCR检测技术，该技术在检测灵敏度和定量方面有显著提高。对8例疑似TBM的连续患者的43份CSF样本进行了分析。CSF样本在标准抗结核治疗（ATT）之前和期间采集。QNRT-PCR检测显示，在ATT治疗过程中采集的43份连续CSF样本中有24份呈阳性结果（55.8%）。此外，通过QNRT-PCR检测分析的M. Tb细菌细胞（BC）数量逐渐减少，这与患者临床状况的改善相关。由于QNRT-PCR检测能够计算CSF样本中M. Tb初始BC数量的数值，该方法是评估TBM临床病程中极其有用且先进的技术。
