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埃塞俄比亚南部奥罗米亚地区甘博医院胃肠道症状患者中十二指肠贾第虫、隐孢子虫属和内阿米巴属的检测与分子特征分析

Detection and molecular characterisation of Giardia duodenalis, Cryptosporidium spp. and Entamoeba spp. among patients with gastrointestinal symptoms in Gambo Hospital, Oromia Region, southern Ethiopia.

作者信息

Flecha María J, Benavides Cynthia M, Tissiano Gabriel, Tesfamariam Abraham, Cuadros Juan, de Lucio Aida, Bailo Begoña, Cano Lourdes, Fuentes Isabel, Carmena David

机构信息

Hospital Príncipe de Asturias, Alcalá de Henares, Madrid, Spain.

Gambo Rural General Hospital, Shasehmane, Ethiopia.

出版信息

Trop Med Int Health. 2015 Sep;20(9):1213-1222. doi: 10.1111/tmi.12535. Epub 2015 May 19.

Abstract

OBJECTIVES

To assess the prevalence and genetic diversity of the enteric protozoa species G. duodenalis, Cryptosporidium spp. and Entamoeba histolytica in individuals with gastrointestinal symptoms compatible with infections by these pathogens seeking medical attention in a rural area in southern Ethiopia.

METHODS

A total of 92 stool samples were initially screened by direct microscopy and immunochromatography and further confirmed by molecular methods. G. duodenalis-positive samples were molecularly characterised by multilocus genotyping of the glutamate dehydrogenase and β-giardin genes of the parasite. PCR and DNA sequence analysis of the gene encoding the 60-kDa glycoprotein was used for the subtyping of Cryptosporidium isolates. Detection and differential diagnosis of E. histolytica/dispar were conducted by real-time PCR.

RESULTS

PCR-based prevalences were 10.9% for G. duodenalis, 1.1% for Cryptosporidium spp. and 3.3% for Entamoeba spp. Seven (four novel and three known) subtypes of G. duodenalis assemblage B were identified at the GDH locus and 5 (one novel and four known) at the BG locus. A novel variant of C. hominis subtype IbA9G3 was also identified. Two Entamoeba isolates were assigned to E. dispar and an additional one to E. histolytica.

CONCLUSION

Although preliminary, our results strongly suggest that giardiasis, cryptosporidiosis and amoebiasis represent a significant burden in Ethiopian rural population.

摘要

目的

评估在埃塞俄比亚南部农村地区因出现与这些病原体感染相符的胃肠道症状而就医的个体中,肠道原生动物物种十二指肠贾第虫、隐孢子虫属和溶组织内阿米巴的流行情况及遗传多样性。

方法

共92份粪便样本最初通过直接显微镜检查和免疫层析法进行筛查,并进一步通过分子方法进行确认。十二指肠贾第虫阳性样本通过对寄生虫的谷氨酸脱氢酶和β-贾第素基因进行多位点基因分型进行分子特征分析。采用编码60 kDa糖蛋白的基因的PCR和DNA序列分析对隐孢子虫分离株进行亚型分析。通过实时PCR对溶组织内阿米巴/迪斯帕内阿米巴进行检测和鉴别诊断。

结果

基于PCR的流行率分别为:十二指肠贾第虫10.9%,隐孢子虫属1.1%,阿米巴属3.3%。在GDH位点鉴定出7种(4种新的和3种已知的)十二指肠贾第虫B组亚型,在BG位点鉴定出5种(1种新的和4种已知的)。还鉴定出一种人隐孢子虫IbA9G3亚型的新变体。2株阿米巴分离株被鉴定为迪斯帕内阿米巴,另外1株为溶组织内阿米巴。

结论

尽管是初步结果,但我们的结果强烈表明,贾第虫病、隐孢子虫病和阿米巴病给埃塞俄比亚农村人口带来了重大负担。

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