依赖于核酸序列恒温扩增技术快速检测副溶血性弧菌方法的建立  

Establishment of the nucleic acid sequence-based amplification method of detecting Vibrio parahaemolyticus

作　　者：倪鑫[1,2] 王志聪[1] 雷质文 梁成珠 汪东风[1] 姜英辉 王建广[3] 祝素贞 

机构地区：[1]中国海洋大学,山东青岛266003 [2]山东检验检疫局,山东青岛266002 [3]青岛科技大学,山东青岛266042

出　　处：《中国预防兽医学报》2011年第11期882-886,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局科研项目(2011IK225)

摘　　要：为建立副溶血性弧菌的快速检测方法,本研究采用依赖于核酸序列恒温扩增(NASBA)技术,以副溶血性弧菌的tlh基因为扩增的靶基因设计特异性引物和探针,建立可快速扩增副溶血性弧菌的NASBA方法。特异性和灵敏度试验结果表明:该NASBA方法对副溶血性弧菌的最小检出量为5.1×102 cfu/mL,高于普通PCR方法,而且与其他种属的菌无任何交叉反应。此外,本研究将副溶血弧菌扩增产物采用通用型核酸扩增物快速检测板进行检测,实现了特异性强的快速副溶血弧菌的检测。该方法对仪器要求低,具有良好的应用前景。In this study,we adopted a technology based on nucleic acid sequence-based amplification（NASBA）,to establish a rapid test method of Vibrio parahaemolyticus.Specific primers and probes were designed for the tlh target gene of V.parahaemolyticus and the detection method of NASBA was established.Its specificity and sensitivity were tested.The results showed that the sensitivity of the NASBA was 5.1×102 cfu/mL which was higher than that of PCR method and this method had no cross reaction with other species of bacteria.The amplification products of V.parahaemolyticus were rapidly detected by using the universal rapid detection board of nucleic acid amplification product.In conclusion,detecting V.parahaemolyticus with NASBA method was more specific and sensitive than PCR method and has lower instrumental requirement and a broad application.

关 键 词：副溶血性弧菌 检测 依赖于核酸序列恒温扩增 

分 类 号：S852.61[农业科学—基础兽医学]