Establishment and application of multiplex realtime fluorescence quantitative PCR assay for detecting various respiratory pathogens

doi:10.3969/j.issn.10073205.2016.11.016

Abstract

Abstract: [
Abstract ] Objective Todevelopmultiplexreal-timefluorescencequantitativePCR ( MRT-
PCR ) assayfor detecting variousrespiratory pathogens.Methods MRT-PCR assay was
establishedwithkindsofrespiratorypathogenicgenes , selectedbasedontheirrelevantreference
strainsbybioinformaticsanalysis , astargetgenes.Standardproductsandqualitycontrolsamples
werebuilttotestingandanalysisingthesensitivity , specificityandreproducibilityofthissystem.
Clinicalsampleswereusedtoevaluating MRT-PCRandsinglefluorescencequantitativePCR.
Results The MRT-PCR detectionsystem weresuccessfullyestablished whichshowedfine
specificity.Thedetectionlimitoftheassayswas10copies / μ L.Coefficientofvariationwasfrom
0. 99%to2. 50% , whichconductedtoassessthereproducibilityoftheassays.Among530clinical
specimens , thedetectableratesof MRT-PCRandsinglefluorescencequantitativePCR were
59. 2% ( 314 / 530 ) and61. 1% ( 324 / 530 ), respectively.Conclusion Wedevelopedafast , specific ,
sensitiveand stable MRT-PCR detection assay , which possessed good clinicalapplication
prospect.

Key words: espiratory tract infections, polymerase chain reaction, sensitivity and specificity

DI Hongqin, YANG Yonghui, KANG Lifei, AN Xiaoying, LI Xiaoxia, ZHANG Hui. Establishment and application of multiplex realtime fluorescence quantitative PCR assay for detecting various respiratory pathogens[J]. Journal of Hebei Medical University, doi: 10.3969/j.issn.10073205.2016.11.016.

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Establishment and application of multiplex realtime fluorescence quantitative PCR assay for detecting various respiratory pathogens

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