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目的初步了解临床腹泻患者贾第虫的感染情况及分子特征。方法以2014年5-7月上海市某医院临床腹泻患者为研究对象,收集粪便样本95份,进行卢戈氏碘液染色,用光学显微镜检查贾第虫包囊。采用巢式PCR法扩增贾第虫磷酸丙糖异构酶基因(tpi),扩增产物测序后用BLAST、Clustal X 1.83和MEGA6.0软件进行同源性和系统发育分析。结果形态学观察发现1例腹泻患者感染贾第虫,阳性检出率为1.05%。其粪便样本经卢戈氏碘液染色后显微镜下可见清晰的包囊。对样本进行PCR检测,扩增片段大小约为530 bp,测序结果显示为贾第虫。经序列比对分析,并以tpi基因构建系统发育树,分析鉴定该分离株为集聚体B,与已报道的人源贾第虫分离株(KF271445)同源性为100%。结论临床腹泻患者中存在贾第虫感染,本研究结果可为了解贾第虫基因型特点及贾第虫病流行病学研究提供参考。
Objective To understand the infection status and molecular characteristics of Giardia in patients with clinical diarrhea. Methods From May to July in 2014, a clinical diarrhea patient was collected in a hospital in Shanghai. 95 stool samples were collected and stained with Lugol’s iodine solution. The cysts of Giardia were examined by light microscopy. Giardia triosomal triose phosphate isomerase gene (tpi) was amplified by nested PCR. The amplified products were sequenced and analyzed by BLAST, Clustal X 1.83 and MEGA 6.0 software for homology and phylogenetic analysis. Results Morphological observation showed that one case of Diarrhea patients infected with Giardia, the positive detection rate was 1.05%. The stool samples by Lugol’s iodine staining after a clear cyst visible capsule. PCR detection of the sample, the amplified fragment size of about 530 bp, the sequencing results showed Giardia. The phylogenetic tree was constructed by tpi gene analysis and sequence analysis. The isolate was identified as agglutinant B. The homology was 100% with the reported isolate of Giardia gondii (KF271445). Conclusion Giardia infection exists in patients with clinical diarrhea. The results of this study may provide reference for understanding the genotype of Giardia and the epidemiology of Giardia.