Williams D L, Spring L, Gillis T P, Salfinger M, Persing D H
Molecular Biology Research Department, Gillis W. Long Hansen's Disease Center at Louisiana State University, Baton Rouge 70894, USA.
Clin Infect Dis. 1998 Feb;26(2):446-50. doi: 10.1086/516313.
In a double-blind study, 655 sputum specimens were obtained from individuals suspected of having tuberculosis and were analyzed for the presence of Mycobacterium tuberculosis and rifampin susceptibility with use of a polymerase chain reaction (PCR)-based universal heteroduplex generator assay (PCR/UHG-Rif). Of the specimens containing viable M. tuberculosis, 100% of the smear-positive (n = 41) and 50% of the smear-negative (n = 6) specimens tested positive for the organism by PCR/UHG-Rif. Nineteen of 537 culture-negative specimens tested positive for M. tuberculosis by PCR/UHG-Rif and were from patients with confirmed tuberculosis who were receiving antituberculosis therapy at the time of specimen collection. Thirty-five specimens contained nontuberculous mycobacteria and were negative by PCR/UHG-Rif. Genotypic evidence of rifampin resistance in five of six culture-confirmed, rifampin-resistant isolates was obtained by PCR/UHG-Rif, yielding a sensitivity and specificity for the assay of 83% and 98.2%, respectively. These results demonstrate the feasibility of using a PCR-based assay directly on sputum specimens for simultaneous detection of M. tuberculosis and rifampin susceptibility, and they suggest that patients with smear-positive, untreated tuberculosis and those presenting with suspected drug-resistant tuberculosis are the most appropriate groups for testing by PCR/UHG-Rif.
在一项双盲研究中，从疑似患有结核病的个体中获取了655份痰标本，并使用基于聚合酶链反应（PCR）的通用异源双链体生成测定法（PCR/UHG-Rif）分析了结核分枝杆菌的存在情况及利福平敏感性。在含有活结核分枝杆菌的标本中，涂片阳性标本（n = 41）通过PCR/UHG-Rif检测该菌的阳性率为100%，涂片阴性标本（n = 6）的阳性率为50%。537份培养阴性标本中有19份通过PCR/UHG-Rif检测结核分枝杆菌呈阳性，这些标本来自确诊为结核病且在采集标本时正在接受抗结核治疗的患者。35份标本含有非结核分枝杆菌，通过PCR/UHG-Rif检测为阴性。在6株培养确诊的利福平耐药菌株中，有5株通过PCR/UHG-Rif获得了利福平耐药的基因型证据，该测定法的敏感性和特异性分别为83%和98.2%。这些结果证明了直接对痰标本使用基于PCR的测定法同时检测结核分枝杆菌和利福平敏感性的可行性，并且表明涂片阳性、未经治疗的结核病患者以及疑似耐多药结核病患者是通过PCR/UHG-Rif进行检测的最合适人群。
