用PCR扩增tim基因检测蓝氏贾第鞭毛虫

采用聚合酶链反应 (PCR)对蓝氏贾第鞭毛虫 (Giardialamblia)的磷酸丙糖异构酶 (triosephosphateisomerase ,缩写为tim)基因进行特异性扩增 ,结果扩增出 1条 6 83bp的DNA片段。此方法的特异性可高达10 0 % ,而其它DNA样本 ,如日本血吸虫 (Schistosomajaponicum)、刚地弓形虫 (Toxoplasmagondii)、微小隐孢子虫 (Cryptosporidiumparvum)、溶组织内阿米巴 (Entamoebahistolytica)、旋毛虫 (Trichinellaspiralis)和阴道毛滴虫 (Trichomonasvaginalis) ,以及人体血细胞等均未出现扩增反应。本法的敏感性也很高 ,可检测到0 4pg贾第虫包囊的DNA。 13株来自不同地理位置和 或宿主的贾第虫DNA样本在PCR中均各产生 1条长为 6 83bp的目的片段。上述结果表明本实验建立的检测贾第虫的PCR方法有效

PCR DETECTION OF GIARDIA LAMBLIA BY TIM GENE AMPLIFICATION

A polymerase chain reaction (PCR) technique was used to amplify the gene of triose phosphate isomerase ( time gene) to detect Giardia lamblia .A fragment of 683bp could be amplified specifically.The primers showed 100% specificity to the isolates of Giardia lamblia only,no amplifications were found with DNAs from other control samples,such as Cryptosporidium parvum,Schistosoma japonicum,Toxoplasma gondii,Entamoeba histolytica,Trichinella spiralis,Trichomonas vaginalis and cells from human. The test could detect even 0 4pg of genomic DNA from Giardia cysts. In addition DNAs from 13 isolates of Giardia lamblia from different geographic locations and/or hosts could also be amplified by PCR,indicating that PCR in the present experiment was effective.