鸡传染性贫血病毒荧光定量PCR检测方法的建立及其在疫苗污染检测中的应用

doi:10.11843/j.issn.0366-6964.2016.07.020

Abstract

Abstract:

To detect the contamination of Chicken infectious anemia virus (CIAV) in attenuated vaccines for poultry，primers and TaqMan probes were designed and synthesized based on the published CIAV genome sequence，and a real-time quantitative PCR method for detecting the CIAV was established by optimizing the reaction conditions.Results analysis showed that the sensitivity of the qPCR may increase detection to 10 copies•μL－1，and the method was approximately 100-fold higher than the sensitivity of the conventional PCR.This detection method had good specificity，and showed no cross-reactions with ALV，REV and MDV genes，the CV (coefficient of variation) of intra-assay and inter-assay were less than 2%，and also showed that the method had the characteristics of better repeatability.In our experiment，a low dosage of 1 EID50•1 000－1 plumes and in 2 positive (5.1%) of 39 samples were detected for CIAV detection from commercial poultry vaccines by qPCR method.In this study，suggesting that this method provides a high sensitivity，strong specificity and better repeatability system for detection of a low dosage CIAV contamination in attenuated vaccines.

Key words: chicken infectious anemia virus, real-time PCR, live vaccine, contamination

CLC Number:

S852.659.2

REN Zhi-hao，FANG Li-chun，LUAN Huai-biao，LI Yang，WANG Yi-xin，CUI Zhi-zhong，CHANG Shuang，ZHAO Peng. Development and Application of a Quantitative PCR for Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Vaccines[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1459-1464.

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Development and Application of a Quantitative PCR for Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Vaccines

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