Detection and Identification of Mycobacterium Tuberculosis by Nested PCR Assays in Cerebrospinal Fluid Samples from Patients with Suspected Tuberculous Meningitis

我們利用先前研發出來的巢式聚合酵素連鎖反應（Nested PCR）來檢測及鑑定臨床上疑似結核性腦膜炎病患的腦脊髓液中的肺結核分枝桿菌。此方法包括以多倍數的IS6110序列和單數的mtp40為標的核酸。當合併使用時，此方法能檢鑑定腦脊髓液中的肺結核分枝桿菌的存在；同樣的檢體以細菌培養法常無法檢測到。我們分析了11個腦脊髓液檢體；其中五例是來自臨床上疑似結核性腦膜炎病患。除了五例外，其它皆為細菌培養陰性。此巢式聚合酵素連鎖反應結果顯示：三例為肺結核分枝桿菌陽性，一例為牛型結核分枝桿菌陽，而只有一例為肺結核分枝桿菌複合群陰性。其它六例，為來自根據臨床所認定的非肺結核分枝桿菌感的病患。令人訝異的是；其中有蛛網膜下出血（SAH）和抗利尿賀爾蒙分泌失調症（SIADH）的檢體為肺結核分枝桿菌陽性。這些發現支持以結核性腦膜炎可能為這些神經病變的原因之一。因此，此巢式聚合酵素連鎖反應提供神經內科醫生一個除了利用臨床表現、腦脊髓液實驗室生化與微生物分析檢驗以外的一個重要輔助方法。

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並列摘要

We used the nested polymerase chain reaction (PCR) assays developed previously to detect and identify Mycobacterium tuberculosis (M. tuberculosis) in the cerebrospinal fluid (CSF) samples from patients with suspected tuberculous meningitis and non-tuberculous patients. Our nested PCR assays target the multi-copy IS6110 insertion element and the single-copy mtp40 genomic DNA of M. tuberculosis. These assays, when used in combination, allowed us to detect a very low number of M. tuberculosis in the CSF samples, which otherwise would be undetectable by the culture method, and to distinguish M. tuberculosis from M. bovis. We applied these nested PCR assays to analyze eleven CSF samples. Among these, five of them were from patients with suspected tuberculous meningitis but all except one were culture negative. Our results of PCR assays show that three of these five are M. tuberculosis positive, one of which is M. bovis positive, and only one is M. tuberculosis negative. The other six CSF samples were from the clinically diagnosed non-tuberculous patients. Surprisingly, two of these so called non-tuberculous patients, a subarachnoid hemorrhage (SAH) and the syndrome of inappropriate antidiuretic hormone secretion (SIADH), were shown M. tuberculosis positive. This finding supports a long-standing argument that tuberculous meningitis is one of the causes of these neurological diseases. These nested PCR assays thus provide the neurologistis with an important adjunct, in addition to the patient’s clinical presentation and laboratory data, for the diagnosis of tuberculous meningitis.

並列關鍵字

Mycobacterium tuberculosis ； PCR ； tuberculous meningitis

國際替代計量

Detection and Identification of Mycobacterium Tuberculosis by Nested PCR Assays in Cerebrospinal Fluid Samples from Patients with Suspected Tuberculous Meningitis

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