中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 基于toxR和rtxA基因水生动物2种病原弧菌的同步检测方法的建立 海洋湖沼通报 2013 (3) 95-100
Position: Home > Articles > DEVELOPMENT OF A DULPLEX PCR FOR DETECTION OF VIBRO PARAHAEMOLYTICUS ANDV.CHOLERAEBASED ON TOXRAND RTXAGENES Transactions of Oceanology and Limnology 2013 (3) 95-100

基于toxR和rtxA基因水生动物2种病原弧菌的同步检测方法的建立

作  者：
白雪松;张晓君;阎斌伦;毕可然;秦蕾
单  位：
中国矿业大学
关键词：
副溶血弧菌;霍乱弧菌;toxR基因;rtxA基因;双重PCR
摘  要：
本研究选择副溶血弧菌(Vibrio parahaemolyticus)的toxR和霍乱弧菌(Vibrio cholerae)的rtxA基因序列设计2对特异性引物,建立了在同一反应体系中同步检测2种病原菌的双重PCR检测方法,扩增目的片段大小分别为368bp和417bp。特异性试验结果表明,在同一反应体系下,副溶血弧菌和霍乱弧菌扩增出目的条带,其他4种对照病原细菌无任何扩增条带;单一及双重PCR敏感性试验结果表明,稀释浓度在(109～102)CFU/mL,单一PCR与双重PCR均可扩增出目的条带,且单一PCR与双重PCR表现出了相同的灵敏度(102 CFU/mL),表明本实验优化的双重PCR反应条件良好;人工染菌样品均可扩增出两条目的条带,表明该方法可直接用于针对病原副溶血弧菌和霍乱弧菌感染的水生动物疾病的检测以及水产品的安全检测,在水产品致病菌的风险评估和大规模样本的分析检测领域具有较高的潜在应用价值。
译  名：
DEVELOPMENT OF A DULPLEX PCR FOR DETECTION OF VIBRO PARAHAEMOLYTICUS ANDV.CHOLERAEBASED ON TOXRAND RTXAGENES
作  者：
BAI Xuesong;ZHANG Xiaojun;YAN Binlun;BI Keran;QIN Lei;China University of Mining and Technology;College of Ocean,Huaihai Institute of Technology;
关键词：
Vibrio parahaemolyticus;;Vibrio cholerae;;toxR gene;;rtxA gene;;Dulplex PCR
摘  要：
Two pairs of specific primers were designed based on toxRgene of Vibro parahaemolyticus and rtxA gene of V.cholerae.A dulplex polymerase chain reaction(PCR)that can simultaneously detect V.parahaemolyticus and V.cholerae was established,and the amplified fragments were 368bp and 417bp,respectively.The results showed that two PCR primers could amplify two gene fragments from chromosomal DNA of V.parahaemolyticus and V.choleraein one PCR reaction,and no positive reaction was detected in the other 4strains of pathogenic bacteria.The results for sensitivity of single and dulplex PCR showed that the two primers could simultaneously detect V.parahaemolyticus and V.cholerae at the level of 109CFU/mL to 102CFU/mL,and the sensitivity of single PCR was as same as that of dulplex PCR.It was proved that the dulplex PCR was an effective assay for detecting V.parahaemolyticus and V.cholerae.Two objective fragments were amplified from artificial infected samples,the results showed that the dulplex PCR protocol could be used in rapid diagnose of aquatic animal diseases caused by V.parahaemolyticus and V.cholerae and safety detection of aquatic food.

相似文章

导出引用
计量
文章访问数: 13
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46