犬瘟热病毒、犬细小病毒二重TaqMan MGB探针实时荧光定量PCR检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2018.08.006

Abstract

Abstract:

This study was aimed to establish a double TaqMan MGB Real-time PCR assay to simultaneously and specifically detect canine distemper virus (CDV) and canine parvovirus (CPV) in one reaction.Two pairs of specific primers for CDV and CPV,along with two TaqMan MGB probes for each virus were designed in the assay basing on CDV H gene and CPV VP2 gene sequences.The specificity,sensitivity and repetition of the double TaqMan MGB Real-time PCR assay were tested,and 48 samples taken from clinic suspicious CDV and CPV infected canines had been testified by the established double TaqMan MGB Real-time PCR.The results indicated that the doulde TaqMan MGB Real-time PCR assay was successfully established,and the number of standard curve correlation (R²) of CDV and CPV were 0.997 and 0.993,respectively.The specificity of the double TaqMan MGB Real-time PCR assay revealed that amplifications were showed on CDV and CPV samples,but other pathogens and negative controls had no amplifications;The sensitivity of CDV and CPV were both 10 copies/μL.Meanwhile,14 CDV positive samples,19 CPV positive samples and 4 CDV/CPV double positive samples were detected,which were consistent with the results of the sequencing.Therefore,the established double TaqMan MGB Real-time PCR assay had high sensitivity,specificity and flux accurate quantitative,which could be applied to clinical CDV/CPV infection each periods.

Key words: canine distemper virus (CDV); H gene; canine parvovirus (CPV); VP2 gene; double TaqMan MGB Real-time PCR

CLC Number:

S852.65⁺5

ZHAO Xueli, LIU Yi, BAN Fuguo, YAN Ruoqian, WANG Huajun, WANG Shujuan, MA Zhenyuan, WANG Dongfang. Establishment and Application of A Double TaqMan MGB Real-time PCR Assay for Detection of Canine Distemper Virus and Canine Parvovirus[J]. , 2018, 45(8): 2086-2094.

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Establishment and Application of A Double TaqMan MGB Real-time PCR Assay for Detection of Canine Distemper Virus and Canine Parvovirus

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