牛副流感病毒3型3种基因型多重RT-PCR检测方法的建立

doi:10.16431/j.cnki.1671-7236.2018.01.004

Abstract

Abstract:

In order to develop a multiplex RT-PCR method for detecting three genotypes of bovine parainfluenza virus type 3 (BPIV3), according to the hemagglutinin-neuraminidase protein genomic sequences of three BPIV3 genotypes obtained from GenBank, three pairs of specific PCR primers were designed. A multiplex RT-PCR detecting three genotypes of BPIV3 was established and optimized. The results showed no cross-reactivity with IBRV, BRSV, BVDV, M.bovis, PPRV, B.melitensis, B.abortus, bovine P.multocida serotype A and bovine P.multocida serotype B. In this study, the amplification produced a series of specific fragments with lengths of 150 bp (BPIV3a), 253 bp (BPIV3b) and 342 bp(BPIV3c), respectively. The limit detection of three recombinant plasmids were 0.89×10⁴, 0.92×10⁴ and 1.53×10⁴ copies/μL. This multiplex RT-PCR method was high specificity and simplicity of operator. It was applied to detect clinical samples and could rapidly detect three genotypes of BPIV3.

Key words: bovine parainfluenza virus type 3 (BPIV3); genotypes; multiplex RT-PCR; detection

CLC Number:

S858.23

YANG Fan, SHI Shunli, XU Na, LEI Yu, CHANG Tana, LI Pingan, GUAN Pingyuan. Establishment of a Multiplex RT-PCR Detection Method for Three Genotypes of Bovine Parainfluenza Virus Type 3[J]. , 2018, 45(1): 32-38.

References

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Establishment of a Multiplex RT-PCR Detection Method for Three Genotypes of Bovine Parainfluenza Virus Type 3

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