应用PCR快速诊断小儿结核性脑膜炎的研究

Rapid diagnosis of tuberculosis meningitis in children using polymerase chain reaction

作者报道特异性寡核苷酸引物引导聚合酶链反应（ＰＣＲ）技术检测脑脊液中结核杆菌ＤＮＡ。所用引物来自于编码结核杆菌的６５ＫＤａ蛋白质抗原基因，产生特异的３８３ｂｐ片段。对１０倍连续稀释的人型结核杆菌ＤＮＡ进行ＰＣＲ，可检测出１ｐｇ结核杆菌ＤＮＡ，相当于１０～１００个细菌。应用ＰＣＲ检测４２例结脑脑脊液，阳性率为５７．１４％，直接涂片镜检和细胞培养阳性率分别为７．１４％和１４．２８％；４６例非结脑脑脊液中，３种检测结果均阴性。研究结果表明，ＰＣＲ检测脑脊液中结核杆菌ＤＮＡ有较高的敏感性和特异性，它快速、简便，有望成为一种临床常规检测结核杆菌方法。

The mycobacterium tuberculosis genome was detected in cerebrospinal fluid by poly-merase chain reaction (PCR ),in which the primers came from the gene coding for the 65 KD My-cobacterial antigens of M. tuberculosis,and a 383 bp segment was amplified using Taq polymerase。 Se-rial dilutions of purified M. tuberculosis DNA(from 1 ng to l00 fg)were used as the templates for thePCR。 Tlie primers could detect less than l pg of DNA。 Of 42 cases with tuberculous meningitis(TBM),24 was positive, with PCR, The percentages of smear and culture were 7.14%and l4. 28%respectively, 46 other neurological disorder were negative, with three methods。 The results suggestedthat PCR be more sensitive and specific than those methods of traditional bacteriology for the detectionof TBM。 PCR is a simple and rapid technique,and it could be used to detect M。 tuberculosis in clinicalspecimens.