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通过对国内来源于不同宿主的ZS人株、SH人株、CN猪株、QH绵羊株4个弓形虫虫株,以及国际标准强毒株RH株的致密颗粒抗原(GRA6)基因进行PCR-RFLP分析,首次对中国弓形虫虫株进行了基因分型研究,旨在了解中国弓形虫基因型的分布情况,从而为进一步的分子遗传学研究以及弓形虫病的防制提供资料。PCR-RFLP分析显示两种带型:RH、SH、CN3株弓形虫为一种带型,QH和ZS2株弓形虫为另一种电泳带型。GRA6序列分析结果显示:RH、SH、CN3株弓形虫的GRA6基因序列上的MseΙ酶切位点与国外报道的RH株一致,同属于基因型Ι,为强毒株;QH和ZS2株弓形虫的MseΙ酶切位点与国外报道的弱毒株BEVERLEY株和ME49株一致,同属于基因型Ⅱ,与PCR-RFLP分析结果一致。除MseΙ酶切位点外,中国的几个虫株GRA6序列与GenBankTM注册的RH株及BEVERLEY和ME49两个弱毒株有个别碱基的差异。结果证明中国人源弓形虫包含了Ι、Ⅱ两种基因型,而动物源性弓形虫则因宿主和来源地不同亦有Ι、Ⅱ两个基因型。
The results of PCR-RFLP analysis showed that the GRA6 gene of four Toxoplasma gondii strains, SH strain, CN pig strain, QH sheep strain and RH strain of RH strain from different hosts in China were PCR-RFLP For the first time, the genotyping of Toxoplasma gondii strains in China was conducted to understand the distribution of Toxoplasma gondii genotypes in China and provide information for further molecular genetics studies and the control of toxoplasmosis. PCR-RFLP analysis showed two bands: Toxoplasma gondii RH, SH and CN3 belonged to one band, while QH and ZS2 Toxoplasma gondii belonged to another band. The results of GRA6 sequence analysis showed that the MseI restriction sites of GRA6 gene of RH, SH and CN3 strains were the same as the RH strains reported abroad, belong to genotype Ι, and were virulent strain. Toxoplasma gondii of QH and ZS2 strains The Msel restriction sites were consistent with those reported in the foreign strains of BEVERLEY and ME49, belonging to genotype II, consistent with the results of PCR-RFLP analysis. In addition to MseI restriction sites, several strains of GRA6 in China have individual base differences with GenBankTM registered RH strains and two attenuated strains of BEVERLEY and ME49. The results showed that Chinese human toxoplasma gondii contained two genotypes, Ⅰ and Ⅱ, while Toxoplasma gondii had two genotypes, Ⅰ and Ⅱ, due to different host and origin.