目的:对分离自大理地区的HIV阳性者感染的弓形虫与RH株进行致密颗粒抗原(GRA6)基因的对比分析。方法采用巢式PCR方法对大理HIV阳性者血液样本与RH株基因组进行扩增,选取GRA6基因阳性扩增产物,用MseⅠ内切酶进行酶切电泳成像,并对基因序列进行测定与分析。结果成功扩增出约800bp大小的GRA6基因片段,MseⅠ内切酶内切后得到约600bp和200bp2个条带。测序结果显示,HIV阳性者血样扩增产物与RH株扩增产物的GRA6基因仅存在2个碱基差异:第447对碱基处C变成G、第623对碱基处G变成T,而且在序列中的146bp和690bp处均找到MseⅠ酶切位点(TTAA)。结论初步判断大理地区的HIV阳性者感染的弓形虫基因型与弓形虫RH株一致,同属基因Ⅰ型。
Objective To comparatively analyze Toxoplasma gondii separated from HIV-positive people and RH strain GRA6 gene. Methods By using the nested PCR the amplification of Dali HIV-positive blood samples and RH strains of Toxo-plasma GRA6 genome was performed. The GRA6 gene amplification positive product was selected and the electrophoresis imag-ing was performed by being digested with the Mse I endonuclease and the gene sequences were measured and analyzed. Re-sults The GRA6 gene fragment 800 bp was successfully amplified and about 600 bp and 200 bp bands were got by Mse I. The sequencing results showed that T. gondii GRA6 gene positive samples had 2 nucleotide variation compared with T. gondii strain RH namely 447 base pair at C becoming G and 623 base pair at G becoming T. At 146 bp and 690 bp the Mse I restric-tion sites TTAA were found. Conclusion The preliminary judgment shows that the Dali HIV-positive T. gondii genotype is consistent with RH strain belonging to genotype I.