摘要
本文借助环介导等温扩增技术(LAMP),建立了两种常见鸡免疫抑制病的(鸡传染性贫血病、J亚群白血病)的快速检测方法。采用简易法提取DNA模板,设计特异性LAMP引物,成功扩增出梯形条带,并且对反应体系进行优化,以及特异性试验和灵敏度试验。结果表明LAMP的灵敏度可达10个拷贝,比PCR灵敏度高10倍;且具有特异性,对其他相关鸡病病原检测结果均为阴性。用建立的LAMP方法对临床样品进行检测,同时与PCR方法比较,结果表明LAMP结果基本与PCR相符,但检测的阳性率要比PCR高。操作简便,无需昂贵设备,适用于临床快速诊断。
A Loop-mediated isothermal amplification（LAMP）assay was developed for rapid detection of chicken infectious anaemia and avian leukosis in this strdy.DNA was extracted as model using simple method.Specific LAMP primers were designed and synthesized,and gradient bands were successfully amplified with reaction system optimized and the speaificity and sensitivity tested.The result showed that the sensilivity of LAMP assay reached 10 copies/test,10 times higher than common PCR with good specifitity.37 Clinical samples were comparatively tested by LAMP and PCR assays with identical result,but the sensitivity of LAMP was higher than PCR.Moreover,the LAMP assay was simple in operation and no expensive instruments were required.It is suitable for rapid clinical diagnasis.
出处
《中国动物检疫》
CAS
2009年第5期29-32,共4页
China Animal Health Inspection
基金
北京市自然基金资助(项目编号:5072011)
