| 同时检测猪圆环病毒2型、猪细小病毒和伪狂犬病毒连接酶检测反应-PCR基因芯片检测方法的建立 |
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| 引用本文： | 郭瑶,汪平,董沁芳,程菊会,徐辉,丁先锋,郭江峰,姜永厚.同时检测猪圆环病毒2型、猪细小病毒和伪狂犬病毒连接酶检测反应-PCR基因芯片检测方法的建立[J].中国预防兽医学报,2011,33(7). |
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| 作者姓名： | 郭瑶 汪平 董沁芳 程菊会 徐辉 丁先锋 郭江峰 姜永厚 |
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| 作者单位： | 1. 浙江理工大学 生命科学学院,浙江杭州,310018
2. 浙江省动物疫病预防控制中心,浙江杭州,310020 |
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| 基金项目： | 浙江省科技计划项目，浙江省自然科学基金，浙江省科技攻关重点项目，浙江省生物医学重中之重学科开放基金 |
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| 摘 要： | 为快速、灵敏、准确地同时检测和鉴别猪圆环病毒2型(PCV2)、猪细小病毒(PPV)和伪狂犬病毒(PRV)的方法,本研究采用连接酶检测反应(LDR)-PCR和基因芯片技术建立一种新型检测方法.首先在3种病毒的保守区内分别设计一对LDR探针,两端各连接一段通用序列,依次进行LDR、通用引物荧光标记扩增和芯片杂交,同时比较引物标记和Cy5 -dCTP标记方法的灵敏度.结果表明该方法可以特异地检测PCV2、PPV和PRV3种病毒,而对牛病毒性腹泻病毒、猪传染性胃肠炎病毒、猪流行性腹泻病毒、猪繁殖与呼吸障碍综合征病毒、猪瘟病毒、乙型脑炎病毒、猪圆环病毒1型检测结果均为阴性;对3种病毒的最低检测限少于10个拷贝;Cy5-dCTP标记检测的灵敏度显著高于引物标记.利用建立的方法对41例临床样品进行检测,与普通PCR检测结果符合率为97.6％～100％.该方法的建立为基础研究和临床应用提供了技术平台.
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| 关 键 词： | LDR-PCR 通用芯片 多重检测 猪病毒 |
A universal oligonucleotide microarray based on ligase detection reaction (LDR)-PCR for parallel detection of porcine circovirus type 2,porcine parvovirus and pseudorabies virus |
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| GUO Yao,WANG Ping,DONG Qin-fang,CHENG Ju-hui,XU Hui,DING Xian-feng,GUO Jiang-feng,JIANG Yong-hou.A universal oligonucleotide microarray based on ligase detection reaction (LDR)-PCR for parallel detection of porcine circovirus type 2,porcine parvovirus and pseudorabies virus[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(7). |
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| Authors: | GUO Yao WANG Ping DONG Qin-fang CHENG Ju-hui XU Hui DING Xian-feng GUO Jiang-feng JIANG Yong-hou |
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| Institution: | GUO Yao1,WANG Ping1,DONG Qin-fang1,CHENG Ju-hui1,XU Hui2,DING Xian-feng1,GUO Jiang-feng1,JIANG Yong-hou1 (1. College of Life Sciences,Zhejiang Sci-Tech University,Hangzhou 310018,China,2. Centre of Animal Diesase Control of Zhejiang Province,Hangzhou 310020,China) |
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| Abstract: | To establish a sensitive detection method for porcine circovirus type 2 (PCV2), porcine parvovirus (PPV) and pseudorabies virus (PRV) which cause swine severe reproductive and/or respiratory failure, a novel diagnostic oligonucleotide microarray based on ligase detection reaction PCR (LDR-PCR) was developed in this study. According to alignment of the viral sequences, LDR probes for each virus were designed, which were flanked by universal sequences on both sides of the probes. Through asymmetric PCR enrich... |
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| Keywords: | LDR-PCR universal microarray parallel detection swine viruses |
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