中国兽医科学2017，Vol.47Issue(4)：461-465,5.DOI:10.16656/j.issn.1673-4696.2017.04.009
旋毛虫LAMP检测方法的建立与应用
Establishment and application of loop-mediated isothermal amplification for the detection of Trichinella spiralis
摘要
Abstract
To establish a fast and accurate method for detection of Trichinellaspiralis in the meat product,loop-mediated isothermal amplification (LAMP) was performed with a set of primers specific to internal transcribed spacer Ⅱ gene(ITS-Ⅱ) of this parasite.To test the sensitivity of this method,T.spiralis genomic DNA extracted from muscle larval was diluted by 10 times as a series,followed by detection with LAMP and conventional PCR.Results showed that the sensitivity of LAMP method was 1 000 times higher than that of PCR.To study the specificity of this method,genomic DNAs from T.spiralis,Toxoplasma gondii,Haemonchus contortus,Eimeria tenella,Cysticercus cellulosae and Escherichia coli were used as templates,respectively.The results indicated that the primers used for the LAMP method was specific for T.spiralis.To get mock samples,different amounts of T.spiralis muscle larvae were added to the same weight of murine muscles(without infection of T.spiralis).Then the DNAs were extracted from these samples and detected by LAMP method.It was found that one larva in 2 grams of meat could be detected by LAMP method.To apply the method,110 tongue samples from pigs,chickens,ducks,lambs and cattle were tested by LAMP.No sample showed infection of T.spiralis.In conclusion,the LAMP method for detection of T.spiralis was established,with the advantages of rapid,sensitive,specific and easy to perform.关键词
旋毛虫/ITS-Ⅱ/环介导等温扩增技术/敏感性/特异性Key words
Trichinella spiralis/internal transcribed spacer Ⅱ/loop-mediated isothermal amplification/sensitivity/specificity分类
农业科技引用本文复制引用
黄芸,徐立新,宋小凯,李祥瑞,严若峰..旋毛虫LAMP检测方法的建立与应用[J].中国兽医科学,2017,47(4):461-465,5.基金项目
江苏省自然科学基金项目(BK20141365) （BK20141365）
