目的对云南大理HIV阳性者人源弓形虫c22-8基因进行分析。方法运用巢式PCR技术对HIV阳性者全血标本进行弓形虫c22-8基因的扩增,扩增产物用限制性内切酶BsmAⅠ和MboⅡ酶切鉴定,并对基因序列进行测定与分析。结果 291份HIV阳性者血样中,32份成功扩增出弓形虫c22-8基因,扩增产物酶切后获得约200bp(〈200bp)、约100bp和〈100bp等3条片段。对扩增产物进行测序分析,云南大理人源弓形虫c22-8基因与基因Ⅰ型标准株(RH株)c22-8基因序列一致。结论初步分析云南大理人源弓形虫与弓形虫基因I型标准株一致。
Objective To analyze the c22-8 gene sites in Toxoplasma g ondii infecting humans in Dali, Yunnan Prov- ince. Method Nested PCR was used to amplify the c22-8 gene from blood samples from individuals who were HIV-pos- itive in Dali. The product was cleaved with the restriction enzymes BsmA Ⅰ and Mbo Ⅱ , and gene sequences were deter mined and analyzed. Results In total, 291 blood samples were collected from HIV positive individuals. The c22 8 gene fragment was successfully amplified from 32 samples. The c22 8 gene fragment was subjected to enzyme digestion and produced 3 bands of about 200 bp (%200 bp), 100 bp, and %100 bp. These bands are consistent with those produced by the standard virulent strain. Sequencing indicated that blood samples with the T. gondii c22-8 gene had the same nucleo tide variation as the RH strain of T. gondii. Conclusion Preliminary analysis indicated that the genotype of T. gondii infecting humans in Dali, Yunnan Province is the same as that of the RH strain of T. gondii.