Chen Cheng-Yen, Ballard Ronald C
Laboratory Reference and Research Branch, Division of STD Prevention, National Center for HIV/AIDS, Viral Hepatitis, STD and Tuberculosis Prevention, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Methods Mol Biol. 2012;903:103-12. doi: 10.1007/978-1-61779-937-2_6.
Highly sensitive and specific nucleic acid amplification tests (NAATs) have emerged as the gold standard diagnostic tests for many infectious diseases. Real-time PCR has further refined the technology of nucleic acid amplification with detection in a closed system and enabled multiplexing to simultaneously detect multiple pathogens. It is a versatile, fast, and high-throughput system for pathogen detection that has reduced the risk of PCR contamination, eliminated post-PCR manipulations, and improved the cost-effectiveness of testing. In addition, real-time PCR can be applied to self-collected noninvasive specimens. Here, we describe an in-house developed TaqMan-based real-time multiplex PCR (M-PCR) assay for the diagnosis of sexually transmitted genital ulcer disease (GUD) and discuss briefly on issues associated with validation of assay performance.
高灵敏度和特异性的核酸扩增检测（NAATs）已成为许多传染病的金标准诊断检测方法。实时荧光定量聚合酶链反应（Real-time PCR）通过在封闭系统中进行检测，进一步完善了核酸扩增技术，并实现了多重检测，能够同时检测多种病原体。它是一种用于病原体检测的通用、快速且高通量的系统，降低了聚合酶链反应（PCR）污染的风险，省去了PCR后的操作，并提高了检测的成本效益。此外，实时荧光定量聚合酶链反应（Real-time PCR）可应用于自行采集的非侵入性标本。在此，我们描述了一种自行研发的基于TaqMan的实时荧光定量多重聚合酶链反应（M-PCR）检测方法，用于诊断性传播性生殖器溃疡疾病（GUD），并简要讨论了与检测性能验证相关的问题。
