猪瘟病毒和牛病毒性腹泻病毒双重荧光定量PCR检测方法的建立和初步应用

doi:10.11843/j.issn.0366-6964.2023.10.038

Abstract

Abstract: The aim of this study was to establish a rapid, efficient and sensitive assay for the dual detection of classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV) by real-time PCR. Specific primers and probes were designed according to the highly conserved regions of CSFV E2 gene and BVDV 5'UTR gene sequences, respectively, and optimized the reaction conditions and program to establish a dual RT-qPCR method. The results showed that the limit detection of this method was 5 copies·μL^-1, which was about 200 times more sensitive than the traditional PCR. This method with high specificity could distinguish CSFV and BVDV, and had no cross-reactivity with common pig pathogens such as foot-and-mouth disease virus (FMDV) and pseudorabies virus (PRV). The coefficients of variations of intra- and inter-group data were less than 2%, demonstrating the good repeatability of this assay. Finally, 109 clinical samples and 7 commercial bovine sera were detected to compare the dual RT-qPCR assay with the CSFV RT-qPCR assay (GB/T 27540-2011) and BVDV RT-qPCR assay (GB/T 18637-2018). The coincident rate was 100%, and the results also illustrated that the BVDV contamination was existed in some commercial fetal bovine serums. Together, this assay could provide technical support for the differential diagnosis and epidemiological field investigation of CSFV and BVDV.

Key words: classical swine fever virus, bovine epidemic diarrhoea virus, dual real-time RT-PCR

CLC Number:

S852.659.6

ZOU Hong, XIA Yingju, LI Ling, XU Lu, ZHAO Junjie, WANG Tuanjie, ZHANG Qianyi, SONG Zhenhui. Establishment and Preliminary Application of a Dual Real-time RT-PCR Assay for CSFV and BVDV[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4422-4427.

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Establishment and Preliminary Application of a Dual Real-time RT-PCR Assay for CSFV and BVDV

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