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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800 bp大小的GRA6基因片段,MseⅠ内切酶内切后得到约600 bp和200 bp 2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146 bp和690 bp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 compare the expression of the granulocyte antigen antigen (GRA6) between Toxoplasma gondii and RH strains infected with HIV-positive individuals isolated in Dali area. Methods The nested PCR method was used to amplify the genomes of RH positive and HIV blood samples from Dali patients. The products of positive amplification of GRA6 gene were selected and digested with Mse Ⅰ endonuclease. The gene sequence was determined and analyzed. RESULTS: The GRA6 gene fragment of about 800 bp in size was successfully amplified. Two fragments of about 600 bp and 200 bp were obtained after Mse Ⅰ endonuclease digestion. Sequencing results showed that there was only 2 bases difference between the amplified product of HIV-positive blood sample and the GRA6 gene of RH strain amplified product: the 447th base changed C to G, the 623th base changed to T, Moreover, the Mse I restriction site (TTAA) was found at 146 bp and 690 bp in the sequence. Conclusion The Toxoplasma gondii genotype infected with HIV-positive in Dali district is consistent with that of Toxoplasma gondii RH strain.