大理某屠宰场猪弓形虫的分离及基因型鉴定

Isolation and Genotype Identification of Toxoplasma gondii from Pigs in Dali,China

为了分离云南省大理猪弓形虫虫株并鉴定其基因型,以大理屠宰场采集到的147份新鲜猪肝为材料,每份猪肝取2g生喂给未感染的实验猫,在喂食病料后6d^8d粪检,获得弓形虫卵囊(命名为TgPDL1)。提取弓形虫卵囊DNA,用套式PCR法扩增弓形虫B1基因并测序,使用PCR-RFLP技术在12个基因位点对弓形虫TgPDL1虫株进行基因型鉴定。结果表明,TgPDL1株扩增片段的序列与NCBI中弓形虫B1基因序列(AF179871)完全相同,12个基因位点与ToxoDB#9型株对比完全一致。说明该虫株为基因型ToxoDB#9型。猪弓形虫大理株的成功分离与鉴定,为进一步研究其生物学特性奠定了基础。

The objective of this study was to isolate T.gondii from pigs in Dali city of Yunnan province and determine the genotypes,147 fresh liver samples were collected from pigs in the local slaughterhouse,then fed an uninfected experimental cat,T.gondii oocysts(named TgPDL1)was obtained after 6-8 days of feeding through stool examination.The oocyst DNA was extracted,the T.gondii B1 gene was amplified by nested PCR and sequenced.DNA from the oocysts were genotyped by PCR/RFLP using 12 genetic markers.Results showed that the sequence of amplified fragment of TgPDL1 strain was the same as the B1 gene(AF179871).The complete genotyping data for all loci indicated the strain belongs to ToxoDB#9.The T.gondii strain was successfully isolated and identified from pigs in Dali.The result provided the foundation for further studying its biological characteristics.