环介导等温扩增技术快速检测志贺菌  

Rapid detection of Shigella by loop-mediated isothermal amplification technology

作　　者：吴家林[1] 沙丹[1] 孙燕萍[1] 马广源[1] 张敬平[1] 凌霞[1] 

机构地区：[1]无锡市疾病预防控制中心,江苏无锡214023

出　　处：《中国卫生检验杂志》2014年第9期1221-1224,共4页Chinese Journal of Health Laboratory Technology

基　　金：卫生部科研基金(WKJ2006-2-10);无锡市卫生系统"十一五"重大综合发展项目(WZD0601);无锡市卫生系统科研项目(XM0901)

摘　　要：目的建立能快速、特异检测志贺菌的环介导等温扩增技术。方法针对志贺菌侵袭性质粒抗原H基因(ipaH)设计4条引物,建立环介导等温扩增检测方法,优化反应体系,测定特异性和灵敏度,与PCR法进行比较,并进行了猪肉模拟样品和实际样品检测。结果环介导等温扩增技术特异性高。灵敏度高于PCR法,对细菌纯培养物的检测灵敏度为101cfu/ml,对猪肉模拟样品检测灵敏度102cfu/g。而PCR法的灵敏度分别为102cfu/ml和104cfu/g。采用环介导等温扩增技术在34份实际样品中检测出5份ipaH基因阳性。结论初步建立能快速、灵敏、特异地检测志贺菌的环介导等温扩增技术。其在食源性疾病监测和应急检测中具有重大意义,值得推广应用。Objective To establish the loop- mediated isothermal amplification（LAMP） technique for rapid detection of Shigella. Methods Four primers were designed specifically according to ipaH gene of Shigella to develop and optimize the LAMP system. The specificity and sensitivity of this system were determined and compared with the polymerase chain reaction（PCR）. Artificially- contaminated pork and practical samples were detected by LAMP to test the practicability of LAMP. Results LAMP technology was highly specific,and showed higher sensitivity than the PCR assay. The sensitivity of the LAMP detection was 10^1cfu / ml for bacteria pure culture samples and 10^2cfu / g for Shigella in artificially- contaminated pork,while those of the PCR assay were 10^2cfu / ml and 10^4cfu / g. ipaH gene positive were detected in 5 samples of 34 practical samples by LAMP technology. Conclusion A rapid,specific and sensitive LAMP technique for detection of Shigella has been preliminarily established,and it is worthy of popularization,with important significance in monitoring and emergency detection of food- borne disease.

关 键 词：环介导等温扩增 志贺菌 IPAH基因 

分 类 号：R446.5[医药卫生—诊断学]