| 罗非鱼源无乳链球菌双重PCR快速检测方法的建立 |
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| 引用本文： | 王均,汪开毓,肖丹,黄锦炉,付希,王浩丞,陈德芳,耿毅,阳涛.罗非鱼源无乳链球菌双重PCR快速检测方法的建立[J].中国兽医科学,2011(5). |
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| 作者姓名： | 王均 汪开毓 肖丹 黄锦炉 付希 王浩丞 陈德芳 耿毅 阳涛 |
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| 作者单位： | 四川农业大学鱼病研究中心;动物疫病与人类健康四川省重点实验室;通威股份有限公司; |
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| 基金项目： | 教育部“长江学者和创新团队发展计划”创新团队项目(IRT0848); 通威股份有限公司重点资助项目(2006-2009) |
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| 摘 要： | 根据GenBank中罗非鱼源无乳链球菌荚膜多糖的cpsF基因和CAMP因子的cfb基因序列,设计2对特异性引物,通过对反应体系和反应条件优化,并进行特异性试验、敏感性试验及人工感染罗非鱼组织样品检测,建立了快速检测无乳链球菌的双重PCR方法。结果显示,仅无乳链球菌能扩增出cpsF和cfb两条特异性条带,而海豚链球菌、金黄色葡萄球菌、鮰爱德华氏菌、嗜水气单胞菌、杀鲑气单胞菌、豚鼠气单胞菌、嗜麦芽寡养单胞菌无条带检出;经敏感性试验,最小模板检出量为7.632×10-2ng/μL;同时对30尾人工感染无乳链球菌的罗非鱼肝和肾组织中细菌DNA进行双重PCR检测和细菌分离鉴定,检出的阳性结果一致。证实,所建立的方法具有快速、特异、灵敏的优点,适用于对罗非鱼等无乳链球菌的感染病例进行流行病学监测。
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| 关 键 词： | 无乳链球菌 cpsF基因 cfb基因 双重聚合酶链反应 罗非鱼 |
Development of double PCR for rapid detection of Streptococcus agalactiae isolated from tilapia |
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| WANG Jun,WANG Kai-yu,XIAO Dan,HUANG Jin-lu,FU Xi,WANG Hao-cheng,CHEN De-fang,GENG Yi,YANG Tao.Development of double PCR for rapid detection of Streptococcus agalactiae isolated from tilapia[J].Veterinary Science in China,2011(5). |
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| Authors: | WANG Jun WANG Kai-yu XIAO Dan HUANG Jin-lu FU Xi WANG Hao-cheng CHEN De-fang GENG Yi YANG Tao |
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| Institution: | WANG Jun1,2,WANG Kai-yu1,XIAO Dan3,HUANG Jin-lu1,FU Xi1,WANG Hao-cheng1,CHEN De-fang1,GENG Yi1,YANG Tao3(1.Fisheries Department,Sichuan Agricultural University,Ya'an 625014,China,2.Key Laboratory of Animal Disease and Human Health of Sichuan Province,3.Tongwei Co.Ltd.,Chengdu 610041,China) |
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| Abstract: | A double PCR method for rapid detection of Streptococcus agalactiae was established.Two pairs of specific primers based on cpsF gene of tilapia S.agalactiae capsular polysaccharides and cfb gene of factor CAMP published in GenBank were designed respectively to perform PCR with optimization of reaction system and reaction condition.In results,two specific bands,cpsF and cfb,were only detected in the S.agalactiae isolates,and none was amplified from Streptococcus iniae,Staphylococcus aureus,Edward-siella icta... |
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| Keywords: | Streptococcus agalactiae cpsF gene cfb gene double PCR tilapia |
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