一种通用型肠道病毒微滴式数字PCR检测方法的建立及应用

doi:10.13217/j.scjpm.2019.0015

Abstract

Abstract: Objective To establish a quantitative detection method of droplet digital polymerase chain reaction （ddPCR） for identification and quantification of enterovirus. Methods The probe concentration and annealing temperature in ddPCR were optimized using enterovirus standards, and the detection range of ddPCR was determined. Viral load assays were performed on 28 clinical samples using optimized reaction conditions. Results In this study, the optimal probe concentration for ddPCR was 0.4 μmol/L, the annealing temperature was 51 °C, the nucleic acid detection range was 3.02-3.59×10⁶ copies/μL, and the detection limit was 3.02 copies/μL. The linear correlation coefficient of the ddPCR method was 0.993 8, showing a good linear relationship. Conclusion This study established a quantification method based on ddPCR for general enterovirus, which can effectively quantify enterovirus load in clinical samples and provide a technique for the determination of viral load in clinical research.

Key words: Digital PCR, Enterovirus, Real-time fluorescence quantitative PCR

CLC Number:

R117

YUAN Run-yu, ZENG Han-ri, SU Juan, LI Wei, MO Yan-ling, LU Jing, KE Chang-wen. Establishment and application of universal droplet digital polymerase chain reaction method for quantification of enterovirus[J].South China Journal of Preventive Medicine, 2019, 45(1): 15-20.

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Establishment and application of universal droplet digital polymerase chain reaction method for quantification of enterovirus

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