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目的了解和比较安徽农村地区人类免疫缺陷病毒(human immunodeficiency virus,HIV)感染者和幼儿园儿童中蓝氏贾第鞭毛虫(Giardia lamblia)感染情况及其基因型。方法2015年4月24日至5月9日收集安徽农村地区某艾滋病治疗点登记在册的HIV感染者及当地一家幼儿园所有儿童的相关信息,并采集粪样进行碘液染色及镜检。提取镜检阳性者的粪样DNA,采用巢式PCR扩增蓝氏贾第鞭毛虫磷酸丙糖异构酶(triosephosphate isomerase,tpi)基因并测序,通过BLAST、Clustal X 1.83和MEGA 6.0软件对基因序列进行同源性比较和系统发育分析。结果共调查HIV感染者127人和儿童125人,发现贾第鞭毛虫感染者4例,其中儿童3例,HIV感染者1例,感染率分别为2.40%(3/125)和0.79%(1/127),两者差异无统计学意义(P>0.05)。3例儿童感染者粪便为成形软便,无腹泻、腹痛等临床症状;1例HIV感染者粪便为稀便,且有明显腹痛、腹泻、乏力症状,体重下降明显。PCR检测结果显示,4例感染者粪样DNA扩增产物均在约500 bp处出现特异条带。测序结果显示,4例均感染蓝氏贾第鞭毛虫,其中HIV感染者2个重复样品的序列相似性为79%,与已报道的上海人源贾第鞭毛虫分离株(Gen Bank登录号:KF271445)相似性分别为79%、98%;3名儿童感染者的样本序列相似性达99%,与上海人源贾第鞭毛虫分离株相似性均为79%。以tpi基因构建的系统进化树结果显示,HIV感染者的蓝氏贾第鞭毛虫分离株为复合型集聚体A+B,3名儿童感染者分离株均为集聚体A;HIV感染者的集聚体A与3名儿童感染者分离株的同源性较高。结论安徽农村地区HIV感染者和儿童人群中存在蓝氏贾第鞭毛虫感染,HIV感染者的虫株基因型为复合型集聚体A+B,儿童的虫株基因型皆为集聚体A。
Objective To understand and compare Giardia lamblia infections and genotypes among HIV-infected and kindergarten children in rural Anhui province. Methods From April 24, 2015 to May 9, 2015, all HIV-infected patients registered in an AIDS treatment center in rural Anhui and all the children in a local kindergarten were collected and dung samples were collected for iodine staining and microscopic examination. The stool-like DNA of the positive samples was extracted. The nested PCR was used to amplify and sequence the gene of triosephosphate isomerase (tpi) of G. lamblia. The genes of BLAST, Clustal X 1.83 and MEGA 6.0 Sequence homology comparison and phylogenetic analysis. Results A total of 127 HIV-infected and 125 children were enrolled. Four cases of Giardia infection were found, of which 3 cases were children and 1 was HIV-infected. The infection rates were 2.40% (3/125) and 0.79% (1) / 127), the difference was not statistically significant (P> 0.05). Three cases of children infected with feces were soft, no diarrhea, abdominal pain and other clinical symptoms; one case of HIV-infected stool loose stools, and obvious abdominal pain, diarrhea, fatigue symptoms, weight loss significantly. The results of PCR showed that specific DNA bands appeared in about 500 bp of DNA amplification products from 4 infected individuals. The sequencing results showed that all the 4 cases were infected with Giardia lamblia, and the sequence similarity of two replicates of HIV-infected persons was 79%, which was similar to the reported genomic sequence of Gen Bank Accession No. GenBank: KF271445) were 79% and 98% respectively. The sequence similarity of the three children infected was 99%, which was 79% similar to that of Shanghai human origin. The phylogenetic tree based on the tpi gene showed that the isolate of A. lamblia from HIV-infected patients was complex cluster A + B, and the isolates of three children infected were all cluster A. Cluster of HIV-infected individuals Body A and three children infected with isolates of high homology. CONCLUSION: Giardia lamblia infection is found in HIV-infected and pediatric populations in rural Anhui. The genotype of HIV-infected strains is A + B and the genotypes of children are all aggregates.