Yam W C, Yuen K Y, Seto W H
Department of Microbiology, The University of Hong Kong, Queen Mary Hospital, Pokfulam.
Clin Chem Lab Med. 1998 Aug;36(8):597-9. doi: 10.1515/CCLM.1998.104.
The performance of an automated DNA amplification assay (Roche Cobas Amplicor Mycobacterium Tuberculosis Test (aPCR) was compared with an in-house single tube nested polymerase chain reaction (nPCR) for the direct detection of Mycobacterium tuberculosis in respiratory specimens. Among 385 specimens, 56 were culture positive for mycobacteria (44 positive for Mycobacterium tuberculosis and 12 positive for non-tuberculosis mycobacteria). The diagnostic sensitivities of aPCR and nPCR were 86% and 91% whereas a 100% diagnostic specificity of both assays was attained. By aPCR, inhibitors were detected in 6% of the clinical samples. For nPCR, the usage of a new thermostable DNA polymerase facilitated pre-PCR decontamination using uracil-N-glycosylase and "hot start" in single step procedure. The results of the study indicated that DNA amplification assays, either manual or automated, were rapid and specific tools for diagnosing pulmonary tuberculosis.
将一种自动化DNA扩增检测方法（罗氏Cobas Amplicor结核分枝杆菌检测（aPCR））与一种用于呼吸道标本中结核分枝杆菌直接检测的内部单管巢式聚合酶链反应（nPCR）进行了比较。在385份标本中，56份分枝杆菌培养呈阳性（44份结核分枝杆菌阳性，12份非结核分枝杆菌阳性）。aPCR和nPCR的诊断敏感性分别为86%和91%，而两种检测方法的诊断特异性均达到100%。通过aPCR，在6%的临床样本中检测到抑制剂。对于nPCR，使用一种新的热稳定DNA聚合酶有助于在单步程序中使用尿嘧啶-N-糖基化酶进行PCR前净化和“热启动”。研究结果表明，DNA扩增检测方法，无论是手动还是自动化的，都是诊断肺结核的快速且特异的工具。
