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通过扩增胸水和胃液中的分枝杆菌DNA检测结核分枝杆菌:IS6110基因与Amplicor分枝杆菌检测法的比较

[Detection of Mycobacterium tuberculosis by amplification of mycobacterial DNA from pleural fluid and gastric juice: the comparison of IS6110 gene with Amplicor mycobacterium].

作者信息

Fusegawa H, Miyachi H, Ohshima T, Satoh T, Ohta S, Fujita K, Ando Y

机构信息

Department of Clinical Pathology, Tokai University School of Medicine, Isehara.

出版信息

Rinsho Byori. 1995 Sep;43(9):941-7.

PMID:7474458
Abstract

The polymerase chain reaction (PCR) using oligonucleotides based on the repetitive sequence (IS6110) of Mycobacterium tuberculosis and Amplicor Mycobacterium, which combines a PCR with the hybridization assay, were evaluated for detection of M. tuberculosis in pleural fluid and gastric juice. The detection limits of these two assay systems for cultured M. tuberculosis were less than 2 cells/ml, as compared with 200 cells/ml by culture. A total of 42 pleural fluid and 94 gastric juice specimens were examined. A total of 5 pleural fluid and 5 gastric juice were culture positive for M. tuberculosis. Only the PCR gave positive results in 2 pleural fluid of which laboratory findings are characteristic of tuberculous pleuritis, and in one gastric juice of the patient who was diagnosed as having pulmonary tuberculosis. Sensitivity, specificity and positive predictive value for IS6110 gene were 75%, 94% and 71.1%, respectively, in pleural fluid. Two of three positive specimens for IS6110 gene from pleural fluid were negative for Amplicor Mycobacterium. These specimens resulted in positive with Amplicor Mycobacterium when solvent extracted DNA was used. Both the PCR systems had the same sensitivity (80%), specificity (98.8%) and positive predictive value (83.3%) in gastric juice. The PCR systems would be useful for rapid detection of M. tuberculosis without long term culture and for detection of non-cultured one from pleural fluid as well as gastric juice. However, when the presence of an inhibitor of PCR is suspected in a specimen, DNA should be purified from the specimen before amplification.

摘要

使用基于结核分枝杆菌重复序列(IS6110)的寡核苷酸进行的聚合酶链反应(PCR)以及结合了PCR与杂交检测的Amplicor分枝杆菌检测法,被用于检测胸腔积液和胃液中的结核分枝杆菌。与培养法检测限为200个细胞/毫升相比,这两种检测系统对培养的结核分枝杆菌的检测限均小于2个细胞/毫升。共检测了42份胸腔积液和94份胃液标本。共有5份胸腔积液和5份胃液结核分枝杆菌培养呈阳性。只有PCR在2份具有结核性胸膜炎实验室特征的胸腔积液以及1份被诊断为肺结核患者的胃液中给出了阳性结果。在胸腔积液中,IS6110基因的敏感性、特异性和阳性预测值分别为75%、94%和71.1%。胸腔积液中IS6110基因的3份阳性标本中有2份Amplicor分枝杆菌检测为阴性。当使用溶剂提取的DNA时,这些标本Amplicor分枝杆菌检测呈阳性。两种PCR系统在胃液中的敏感性(80%)、特异性(98.8%)和阳性预测值(83.3%)相同。PCR系统对于无需长期培养快速检测结核分枝杆菌以及检测胸腔积液和胃液中未培养的结核分枝杆菌将很有用。然而,当怀疑标本中存在PCR抑制剂时,应在扩增前从标本中纯化DNA。

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