通过实时逆转录聚合酶链反应检测和定量基孔肯雅病毒

Detection and Quantification of Chikungunya Virus by Real-Time RT-PCR Assay.

作者信息

Wang Seok Mui, Ali Ummul Haninah, Sekaran Shamala Devi, Thayan Ravindran

机构信息

Faculty of Medicine, Jalan Hospital, Universiti Teknologi MARA (UiTM), Sungai Buloh Campus, 47000 Sungai Buloh, Selangor Darul Ehsan, Malaysia.

Department of Medical Microbiology, Faculty of Medicine, University of Malaysia, Jalan Universiti, Kuala Lumpur, 50603, Malaysia.

出版信息

Methods Mol Biol. 2016;1426:105-17. doi: 10.1007/978-1-4939-3618-2_10.

DOI:10.1007/978-1-4939-3618-2_10

PMID:27233265

Abstract

Real-time PCR assay has many advantages over conventional PCR methods, including rapidity, quantitative measurement, low risk of contamination, high sensitivity, high specificity, and ease of standardization (Mackay et al., Nucleic Acids Res 30:1292-1305, 2002). The real-time PCR system relies upon the measurement of a fluorescent reporter during PCR, in which the amount of emitted fluorescence is directly proportional to the amount of the PCR product in a reaction (Gibsons et al., Genome Res 6:995-1001, 1996). Here, we describe the use of SYBR Green I-based and TaqMan(®) real-time reverse transcription polymerase chain reaction (RT-PCR) for the detection and quantification of Chikungunya virus (CHIKV).

摘要

与传统PCR方法相比，实时PCR检测具有许多优势，包括快速性、定量测量、低污染风险、高灵敏度、高特异性以及易于标准化（Mackay等人，《核酸研究》30:1292 - 1305，2002年）。实时PCR系统依赖于在PCR过程中对荧光报告分子的测量，其中发射的荧光量与反应中PCR产物的量直接成正比（Gibsons等人，《基因组研究》6:995 - 1001，1996年）。在此，我们描述基于SYBR Green I和TaqMan®实时逆转录聚合酶链反应（RT-PCR）用于检测和定量基孔肯雅病毒（CHIKV）。