摘要
为了建立一种能快速检测衣阿华支原体(MI)的方法,根据基因库中MI种蛋白基因(species pro-tein 1)的保守序列,设计了1套特异性环介导等温扩增(LAMP)引物,建立了用以可视化检测衣阿华支原体的LAMP方法。结果显示,用此LAMP方法对衣阿华支原体的检测结果为阳性(肉眼观察可见翠绿色),而对其他常见病原体的检测结果均为阴性(肉眼观察可见桔红色),敏感性可达10fg DNA,高于常规PCR 100倍;该方法可在1h内完成,可通过肉眼观察颜色直接判定结果。结果表明,建立的LAMP方法简便、快速、灵敏、特异,可用于禽MI感染的快速检测。
A loop-mediated isothermal amplification（LAMP） assay was developed for detection of Mycoplasma iowae（MI）.According to the sequences specific to species protein1 of MI in GenBank,six primer pairs were designed,and the reaction conditions were optimized. In result,only LAMP reactions of Mycoplasma iowae were positive（visible green）,while LAMP reactions of the other common pathogens were negative（visible red）.The detection limit of this LAMP method was 10 fg for DNA,which was 100 times higher than routine PCR.The amplification could be finished within 1 h,and the presence of MI could be detected by naked eyes.These results show that this LAMP assay is a simple and specific method for rapid detection of MI for clinical samples.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2013年第8期817-821,共5页
Chinese Veterinary Science
基金
广西特聘专家专项(2011B020)
国家百千万人才工程人选专项(945200603)
广西科技重大专项(1222003-2-4)
关键词
衣阿华支原体
环介导等温扩增
检测
Mycoplasma iowae
loop-mediated isothermal amplification（LAMP）
detection
