作　　者： ; ; ; ; ;

机构地区： 华南农业大学兽医学院

出　　处： 《中国预防兽医学报》 2004年第3期231-233,237,共4页

摘　　要： 经BLAST检索,以HSP70基因设计一对引物(5'＿CAATCGAATTGGATTCTTTGGTC＿3'和5'＿CACCTTCAAAT ACTTGAATAAGT＿3')对奶牛安氏隐孢子虫进行了PCR试验。结果显示所建立的PCR检测方法只能特异扩增隐孢子虫GD株DNA,而对照样本如微小隐孢子虫、弓形虫、圆孢子虫、纤毛虫、肝片吸虫、血矛线虫、莫尼茨绦虫、牛粪便以及大肠杆菌均为阴性;通过对6个浓度梯度的虫体DNA进行PCR反应,结果表明当样本中含有445个隐孢子虫卵囊的DNA时,即可扩增产生清晰可辩的条带。测得该序列长度为494bp,序列分析为牛型C.andersoni。表明该引物能特异扩增C.andersoni,敏感性较高,适合于奶牛安氏隐孢子虫的检测。 To develop a sensitive and species-specific PCR diagnostic technique for detection and identification of Cryptosporidium andersoni oocysts, a pair of primers (the forward primer-5' CAATCGAATTGGATTCTTTGGTC 3' and the reverse primer-5' CACCTTCAAATACTTGAATAAGT 3') were designed based on the published HSP70 gene.Only a 494bp target DNA fragment was amplified from C.andersoni oocyst DNA by the PCR. While no amplification products were detected in the control DNA samples extracted from C. parvum,Eimeria tenella,Escherichia coli,Toxoplasma gondii,Haemonchus contortus, Moniezia,cattle feces,Fasciolopsis hepatica,Balantidium coli.The C. andersoni oocyst DNA fragment amplified was sequenced. Sequence analysis confirmed that the species was C.andersoni. The results indicated that the PCR technique was sensitive and could detect DNA of 445 oocysts of C.andersoni.The PCR technique could be used as a diagnostic tool for the detection and identification of C.andersoni.

关 键 词： 安氏隐孢子虫 种特异性

领　　域： [农业科学] [农业科学] [农业科学]