摘要
目的建立一种可以根据颜色变化判断结果的环介导等温扩增方法(LAMP)检测贝类包拉米虫。方法根据贝类包拉米虫的基因,设计一套LAMP引物,进行LAMP反应,并优化反应条件,进行特异性和敏感性实验。结果经过反应条件优化,可通过肉眼观察颜色直接判定结果,在1h内完成整个反应,检测的敏感性最低达到15.4fg,是普通PCR方法的10倍,对其他贝类常见病原体的检测结果均为阴性。结论本实验建立的检测贝类包拉米虫的LAMP方法,具有良好的特异性和敏感性,不需要昂贵的仪器,可以在短时间内检测包拉米虫,在基层兽医具有良好的诊断应用价值。
Objective To develop a loop-mediated isothermal amplification （LAMP） assay for rapid detection of Bonamia based on the colour change. Method A set of specific primers targeting the conservative region of Bonamia was designed for LAMP assay, the reaction parameters were optimized and then specificity and sensitivity were tested. Result The results could be observed within an hour after optimization of the reaction conditions. The detection limit of this LAMP assay was 15.4 fg, 10 times sensitive than routine PCR assay and had no cross reaction with other oyster pathogens. Conclusion The LAMP assay was of good specificity and sensibility, and results could be observed in a short time without expensive machines to detect Bonamia from oyster samples, It could applied for Grassroots veterinary clinical diagnosis.
出处
《中国动物检疫》
CAS
2013年第4期43-46,共4页
China Animal Health Inspection
基金
国家百千万人才工程人选专项资金(945200603)
广西特聘专家专项(2011B020)
广西科技攻关项目(桂科攻0630001-3M)
关键词
包拉米虫
环节导等温扩增
可视化检测
Bonamia. loop-mediated isothermal amplification
visual detection
