猪流行性腹泻病毒与圆环病毒2型复合PCR方法的建立

doi:10.3969/j.issn.1004-1524.2017.07.05

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (7): 1086-1092.DOI: 10.3969/j.issn.1004-1524.2017.07.05

猪流行性腹泻病毒与圆环病毒2型复合PCR方法的建立

龚双燕^a, 李小璟^a, 陈瑛琪^a, 蔡瑶^a, 李雨濛^a, 徐逸飞^a, 朱玲^{a, b}, 徐志文^{a, b, *}

a. 四川农业大学动物医学院,四川成都 611130;
b. 四川农业大学动物疫病与人类健康四川省重点实验室,四川成都 611130

收稿日期:2016-12-27 出版日期:2017-07-20 发布日期:2017-07-24
通讯作者: 徐志文,E-mail:abtcxzw@126.com
作者简介:龚双燕(1994—),女,重庆开县人,硕士研究生,主要从事动物传染病病原分子生物学研究。E-mail:654392857@qq.com
基金资助:
四川省科技支撑计划(2017NZ0038); “十二五”农村领域国家科技计划课题(2015BAD12B04-2.3)

Establishment of a multiplex PCR for porcine epidemic diarrhea virus and porcine circovirus type 2

GONG Shuangyan^a, LI Xiaojing^a, CHEN Yingqi^a, CAI Yao^a, LI Yumeng^a, XU Yifei^a, ZHU Ling^{a, b}, XU Zhiwen^{a, b, *}

a. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China;
b. Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China

Received:2016-12-27 Online:2017-07-20 Published:2017-07-24

摘要/Abstract

摘要： 为研究猪流行性腹泻病毒(PEDV)、猪圆环病毒2型(PCV2)在腹泻仔猪中的感染状况,建立一种能同时检测 PEDV与PCV2混合感染的复合PCR方法。根据GenBank已发表的PEDV、PCV2的基因序列,选择保守区域分别设计、合成1对特异性引物,扩增目的片段分别为530和278 bp。经反应条件优化,建立了特异性检测PEDV、PCV2的复合PCR方法。应用本试验所建复合PCR方法,对2016年4—8月采自四川省乐山、宜宾、广元和遂宁等地区的67份腹泻仔猪样品(肠系膜淋巴结、肠道黏膜及其内容物混合)进行检测。结果显示:PEDV、PCV2单一感染阳性率分别为34.33%、73.13%;PEDV+PCV2复合感染率为16.42%。经与特异性检测PCV2的PCR法和PEDV的RT-PCR法检测结果进行比较分析,符合率均为100%。

关键词: 流行性腹泻病毒, 圆环病毒2型, 复合PCR

Abstract: In order to study the infection status of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 2 (PCV2) in diarrhea piglets, a multiplex PCR method for simultaneous detection of PEDV and PCV2 mixed infection was established. According to the published sequence of PEDV and PCV2 in GenBank, 2 pairs of specific primers were designed and synthesized for the conserved region, and the amplified fragments were 530 and 278 bp, respectively. The multiplex PCR method for the specific detection of PEDV and PCV2 was established by optimizing the reaction conditions. A total of 67 diarrhea samples (mixed mesenteric lymph nodes, intestinal mucosa and their contents) were collected from Leshan, Yibin, Guangyuan and Suining of Sichuan Province from April to August, 2016, and were detected by multiplex PCR. The results showed that the single infection positive rates of PEDV and PCV2 was 34.33% and 73.13%, respectively. The combined infection rate of PEDV and PCV2 was 16.42%. The coincidence rates of the multiplex PCR method and the specific detection methods of PCV2 PCR and PEDV RT-PCR were all 100%.

Key words: porcine epidemic diarrhea virus, porcine circovirus type 2, multiplex PCR

中图分类号:

S855.3

龚双燕, 李小璟, 陈瑛琪, 蔡瑶, 李雨濛, 徐逸飞, 朱玲, 徐志文. 猪流行性腹泻病毒与圆环病毒2型复合PCR方法的建立[J]. 浙江农业学报, 2017, 29(7): 1086-1092.

GONG Shuangyan, LI Xiaojing, CHEN Yingqi, CAI Yao, LI Yumeng, XU Yifei, ZHU Ling, XU Zhiwen. Establishment of a multiplex PCR for porcine epidemic diarrhea virus and porcine circovirus type 2[J]. , 2017, 29(7): 1086-1092.

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猪流行性腹泻病毒与圆环病毒2型复合PCR方法的建立

Establishment of a multiplex PCR for porcine epidemic diarrhea virus and porcine circovirus type 2

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