摘要
为建立一种鸡毒支原体(MG)的快速检测方法,本研究根据GenBank中MG的基因保守序列,设计1套特异性环介导等温扩增(LAMP)引物,优化反应条件,建立了MG LAMP的可视化检测方法。该方法的敏感性可达10fg/μL,高于常规PCR方法100倍;其特异性好,对其他的鸡常见病原体的检测结果均为阴性。MG LAMP反应只需一个可控温的水浴锅,在1h内完成全部反应,通过肉眼观察颜色直接判定结果。本研究建立的LAMP方法具有灵敏度高、特异性好、操作简便的优点,可用于MG感染的快速检测。
A loop-mediated isothermal amplification（LAMP） assay was developed for detection of mycoplasma gallisepticum（MG）.According to the sequences of MG in GenBank,six primers were designed,and the reaction conditions were optimized.The results showed that the detection limit of this LAMP method was 10 fg/μL,which was 102-fold higher than the routine PCR.There was no cross-reactivity with the other pathogen of chicken.The amplification could be finished by water bath within 1 h,and the presence of MG could be detected and visualized by naked eyes.These results suggested that this LAMP assay was a simple and specific method for rapid detection of MG of clinical samples.
出处
《中国畜牧兽医》
CAS
北大核心
2012年第9期30-33,共4页
China Animal Husbandry & Veterinary Medicine
基金
广西特聘专家专项经费(2011B020)
新世纪百千万人才工程国家级人选专项经费(945200603)
