鸡细小病毒半巢式PCR检测方法的建立及初步应用

doi:10.16431/j.cnki.1671-7236.2017.05.006

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (5): 1295-1301.doi: 10.16431/j.cnki.1671-7236.2017.05.006

鸡细小病毒半巢式PCR检测方法的建立及初步应用

奉彬¹, 谢芝勋¹, 张艳芳¹, 黄娇玲¹, 王盛¹, 范晴¹, 黄莉¹, 谢丽基¹, 曾婷婷¹, 罗思思¹, 邓显文¹, 谢志勤¹, 刘加波¹, 宋中宝², 林二克²

1. 广西壮族自治区兽医研究所, 广西兽医生物技术重点实验室, 南宁 530001;
2. 南宁市良凤农牧有限责任公司, 南宁 530031

收稿日期:2016-11-17 出版日期:2017-05-20 发布日期:2017-05-18
通讯作者: 谢芝勋 E-mail:xiezhixun@126.com
作者简介:奉彬(1987-),女,广西贺州人,硕士,研究方向:禽病快速诊断技术,E-mail:fengbin0609@163.com
基金资助:
广西科技项目（15-140-31-A-1、15-140-31-B-1、桂科AB16380003）；南宁市科技攻关项目（20152308）；国家“万人计划”领军人才专项（2016-37-88）

Development and Preliminary Application of a Semi-nested PCR Assay for Detection of Chicken Parvovirus

FENG Bin¹, XIE Zhi-xun¹, ZHANG Yan-fang¹, HUANG Jiao-ling¹, WANG Sheng¹, FAN Qing¹, HUANG Li¹, XIE Li-ji¹, ZENG Ting-ting¹, LUO Si-si¹, DENG Xian-wen¹, XIE Zhi-qin¹, LIU Jia-bo¹, SONG Zhong-bao², LIN Er-ke²

1. Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Nanning 530001, China;
2. Nanning Liang Feng Agriculture and Animal Husbandry Co., Ltd., Nanning 530031, China

Received:2016-11-17 Online:2017-05-20 Published:2017-05-18

摘要/Abstract

摘要：

为建立一种快速、特异、灵敏的检测鸡细小病毒（chicken parvovirus，ChPV）的方法，根据ChPV的保守基因NS1设计了3条特异性引物，建立并优化了能快速检测ChPV的半巢式PCR方法，对其进行特异性和敏感性试验，并用所建立的方法对48份临床样品进行了检测。特异性和敏感性试验结果显示，建立的半巢式PCR只对ChPV敏感，扩增产物为186 bp的特异性条带；其最低能检测到5.62 fg/μL的ChPV DNA；而对鸡新城疫病毒、H9亚型禽流感病毒、马立克氏病病毒、鸡传染性喉气管炎病毒、鸡传染性支气管炎病毒不敏感。临床检测结果显示，同时对48份临床样品进行检测，检出率为16.67%（8/48），提示广西区内鸡群存在ChPV感染。本研究建立的ChPV半巢式PCR方法适用于ChPV的临床检测。

关键词: 鸡细小病毒; 半巢式PCR; 检测

Abstract:

In order to set up and optimize a semi-nested PCR for rapid detection of chicken parvovirus (ChPV), three specific primers were designed according to conserved sequences of NS 1 gene of ChPV. The specificity and sensitivity of ChPV semi-nested PCR were tested, and the assay was applied to detect 48 clinical samples. The specificity and sensitivity tests showed that this semi-nested PCR was only sensitive to ChPV for amplifying specific band of 186 bp and it could detect 5.62 fg/μL of ChPV DNA, without any sensitivity to other viruses, such as Newcastle disease virus, H9 subtype avian influenza virus, Marek's disease virus, infectious laryngotracheitis virus and infectious bronchitis virus. 48 chicken samples were detected and the positive rate was 16.67% (8/48). The results of our study demonstrated that the optimized semi-nested PCR could be a method that was suitable for clinical detection of ChPV.

Key words: chicken parvovirus; semi-nested PCR; detection

中图分类号:

S858.31

奉彬, 谢芝勋, 张艳芳, 黄娇玲, 王盛, 范晴, 黄莉, 谢丽基, 曾婷婷, 罗思思, 邓显文, 谢志勤, 刘加波, 宋中宝, 林二克. 鸡细小病毒半巢式PCR检测方法的建立及初步应用[J]. 《中国畜牧兽医》, 2017, 44(5): 1295-1301.

FENG Bin, XIE Zhi-xun, ZHANG Yan-fang, HUANG Jiao-ling, WANG Sheng, FAN Qing, HUANG Li, XIE Li-ji, ZENG Ting-ting, LUO Si-si, DENG Xian-wen, XIE Zhi-qin, LIU Jia-bo, SONG Zhong-bao, LIN Er-ke. Development and Preliminary Application of a Semi-nested PCR Assay for Detection of Chicken Parvovirus[J]. , 2017, 44(5): 1295-1301.

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鸡细小病毒半巢式PCR检测方法的建立及初步应用

Development and Preliminary Application of a Semi-nested PCR Assay for Detection of Chicken Parvovirus

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