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[从一名哥伦比亚艾滋病患者粪便中首次分离出肠脑炎微孢子虫]

[First isolate of Encephalitozoon intestinalis from stools of a Colombian patient with AIDS].

作者信息

Bedoya Katherine, Montoya Martha Nelly, Botero Jorge, Galván Ana Luz

机构信息

Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales - GIEPI, Corporación para el Estudio de las Patologías Tropicales - CAEPT, Universidad de Antioquia, Medellín, Colombia.

出版信息

Biomedica. 2008 Sep;28(3):441-7.

Abstract

INTRODUCTION

Microsporidia are obligate intracellular parasites that are recognized as important opportunistic pathogens of immunocompromised and transplanted patients. Enterocytozoon bieneusi and, less frequently, Encephalitozoon intestinalis are the most prevalent species in humans; both of them are associated with enteric infections. Cell cultures have been useful in the study of microsporidia biology. In Colombia, however, no isolates of microsporidia from patients with AIDS have been obtained.

OBJECTIVE

A cell culture of intestinal microsporidia was established from stools of positive patients in order to isolate a native strain.

MATERIALS AND METHODS

Stool from a single AIDS patient was concentrated with the water-ether technique, and the sediment was treated with a mixture of antibiotics and antifungal agents for 18 hours at 37 degrees C. Vero cells were cultivated in 24-well plates with Gibco RPMI medium supplemented with 10% bovine fetal serum and antibiotics. The culture was subsequently inoculated with previously concentrated spores. The medium was changed every second day and the presence of spores was evaluated with the Quick Hot Gram chromotrope stain.

RESULTS

Two weeks post-infection, microsporidial spores were identified with characteristic morphology and staining properties. PCR results showed that Encephalitozoon intestinalis was the isolated species.

CONCLUSIONS

A cell culture of microsporidia was established from a stool sample. This protocol is important to isolate and maintain additional native Colombian strains and it will contribute to biochemical, immunological and epidemiological studies of the currently established strain.

摘要

引言

微孢子虫是专性细胞内寄生虫,被认为是免疫功能低下和移植患者重要的机会性病原体。比氏肠微孢子虫是人类中最常见的种类,肠脑炎微孢子虫相对较少见;它们都与肠道感染有关。细胞培养在微孢子虫生物学研究中很有用。然而,在哥伦比亚,尚未从艾滋病患者中获得微孢子虫分离株。

目的

为了分离出本地菌株,从阳性患者的粪便中建立肠道微孢子虫的细胞培养。

材料与方法

用乙醚水技术浓缩一名艾滋病患者的粪便,沉淀物在37℃下用抗生素和抗真菌剂混合物处理18小时。将Vero细胞接种于含10%胎牛血清和抗生素的Gibco RPMI培养基的24孔板中。随后用先前浓缩的孢子接种培养物。每隔一天更换培养基,并用快速热革兰变色染液评估孢子的存在情况。

结果

感染两周后,鉴定出具有特征形态和染色特性的微孢子虫孢子。PCR结果表明分离出的物种是肠脑炎微孢子虫。

结论

从粪便样本中建立了微孢子虫的细胞培养。该方案对于分离和保存更多哥伦比亚本地菌株很重要,并且将有助于对目前已建立的菌株进行生化、免疫和流行病学研究。

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