猪瘟病毒实时荧光定量PCR检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2017.07.029

Abstract

Abstract:

A Real-time quantitative PCR assay for detection of classical swine fever virus (CSFV) was developed using the specific probe and primers designed basing on the E2 gene of CSFV. The Real-time quantitative PCR assay was established using the total RNA of CSFV as template. The specificity, sensitivity and repeatability of the assay were tested, and samples taken from clinic suspicious CSFV infected pigs had been testified by the established assay. The results indicated that the Real-time quantitative PCR assay was successfully established, and showed a good linear relationship at a template range of 10¹ to 10⁶ copies/μL with a coefficient correlation of 0.999; The specificity of the assay revealed that amplifications were showed on CSFV samples, but other pathogens had no amplifications; The sensitivity of the assay was 10 copies/μL nucleic acid and 1 TCID₅₀/mL virus; Meanwhile,19 positive samples were detected, which were consistent with results of CSFV detected by Nested RT-PCR, cloning and sequencing. The eatablished Real-time quantitative PCR assay was specific, sensitive rapid and suitable for early detection and epidemiological study of CSFV.

Key words: classical swine fever virus; Real-time quantitative PCR; specificity; repeatability; sensitivity; application

CLC Number:

S852.65⁺1

WANG Shu-juan, LIU Mei-fen, YAN Ruo-qian, BAN Fu-guo, ZHAO Xue-li, MA Zhen-yuan, WANG Hua-jun, WANG Cui, ZHAO Ming-jun. Establishment and Application of Real-time Quantitative PCR Assay for Detection of Classical Swine Fever Virus[J]. , 2017, 44(7): 2112-2118.

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Establishment and Application of Real-time Quantitative PCR Assay for Detection of Classical Swine Fever Virus

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