Zambardi G, Druetta A, Roure C, Fouqué B, Girardo P, Chypre C, Marchand J, Freney J, Fleurette J
Départment dEtudes et de Recherche en Bactériologie (EA 1655) Faculté de Médecine Alexis Carrel, Lyon, France.
Mol Cell Probes. 1995 Apr;9(2):91-9. doi: 10.1016/s0890-8508(95)80033-6.
A polymerase chain reaction (PCR) assay was developed for the detection in clinical samples of mycobacteria belonging to the Mycobacterium tuberculosis complex. PCR products were detected with a simple and rapid colormetric method. With this method, 50 fg of M. tuberculosis DNA were detectable with the repetitive DNA-sequence-derived primers, corresponding to 10 genome equivalents. Detection of M. tuberculosis in 258 clinical samples by PCR was compared with detection by culture. PCR was positive for 56 of 57 culture-positive and Ziehl-Neelsen-staining-positive (ZN) samples, 11 of 18 culture-positive and ZN-negative samples. The presence of groEL DNA sequences was also investigated by PCR for all the specimens with the same revelation protocol. Three of the eight false-negative samples with the repetitive element-derived primers were found to contain groEL DNA sequences specific for the Mycobacterium genus. Among the 183 culture-negative samples, 30 were positive by PCR. When clinical data were known, the diagnosis of tuberculosis was established for the patients from whom those samples had been obtained. The results show that the rapid and simplified PCR assay described here is slightly more sensitive than culture and can be used in routine clinical practice.
开发了一种聚合酶链反应（PCR）检测方法，用于检测临床样本中属于结核分枝杆菌复合群的分枝杆菌。采用一种简单快速的比色法检测PCR产物。用这种方法，使用重复DNA序列衍生引物可检测到50 fg的结核分枝杆菌DNA，相当于10个基因组当量。将PCR法检测258份临床样本中的结核分枝杆菌与培养法检测结果进行比较。在57份培养阳性且齐-尼氏染色阳性（ZN）的样本中，PCR有56份呈阳性；在18份培养阳性但ZN阴性的样本中，有11份呈阳性。还用相同的显色方案通过PCR对所有标本检测groEL DNA序列的存在情况。在8份使用重复元件衍生引物检测为假阴性的样本中，有3份被发现含有分枝杆菌属特异性的groEL DNA序列。在183份培养阴性的样本中，有30份PCR呈阳性。当了解临床数据时，对获取这些样本的患者确诊为结核病。结果表明，本文所述的快速简化PCR检测方法比培养法稍敏感，可用于常规临床实践。
