作　　者： ; ; ; ; ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《中国兽医科学》 2015年第1期80-84,共5页

摘　　要： 为了对野生动物捻转血矛线虫进行分子鉴定并建立特异性PCR检测方法,基于ITS序列,对草食类野生动物粪便中毛圆线虫虫卵及幼虫DNA进行分子鉴定,并比较粪便和虫体样品的PCR扩增效果;基于ITS1序列建立了特异性PCR检测方法,并对20份临床粪样进行了检测。结果显示,2种草食动物(长颈鹿、弯角剑羚)粪便和虫体样品均扩增出ITS1和ITS2片段,且虫体DNA比虫卵DNA的扩增效果更好;经系统发育分析和BLAST比对,该毛圆线虫分离株属于毛圆科血矛属,与基因库中多株捻转血矛线虫ITS1和ITS2序列的相似性为98%~99%。结果表明,基于ITS1序列建立的PCR检测方法具有良好的特异性和敏感性,与幼虫鉴定法的符合率为95%,高于粪便虫卵检查法(70%),具有一定的临床应用价值。 To identify molecularly Haemonchus contortus from wildlife animals and to establish specific PCR for detecting it,molecular identification of the DNA from eggs and larvae in feces of herbivorous wildlife was conducted respectively based on ITS sequences,and PCR amplification effects between two DNA samples were compared.A PCR for the detection of H.contortus was established based on ITS1 sequence and 20 clinical samples were detected by this assay.In result,ITS1 and ITS2fragments of egg or larval samples from two herbivores including Giraffa cameleopardalis and Oryx dammah were amplified,and larval DNA had better stability and accuracy in PCR amplification.Phylogenetic analysis and BLAST alignment showed that they belonged to Haemonchus of Trichostrongylidae,and the sequence similarity between all isolates from herbivores and the H.contortus references in GenBank was from 98%to 99%at the ITS1 and ITS2loci.The result showed that the established PCR had good specificity and sensitivity,and the accordance rate of the PCR detection with the larval identification was 95%,higher than that of the feces examination(70%).Therefore,it has certain clinical application value.

关 键 词： 捻转血矛线虫 内转录间隔区 分子鉴定 检测 野生动物

领　　域： [农业科学] [农业科学] [农业科学]