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原位杂交:一种用于诊断微孢子虫寄生虫比氏肠微孢子虫的分子方法。

In situ hybridization: a molecular approach for the diagnosis of the microsporidian parasite Enterocytozoon bieneusi.

作者信息

Velásquez J N, Carnevale S, Labbé J H, Chertcoff A, Cabrera M G, Oelemann W

机构信息

Hospital Municipal de Infecciosas Dr. Francisco Javier Muñiz, Instituto Nacional de Microbiologia Dr. Carlos G. Malbrán, Buenos Aires, Argentina.

出版信息

Hum Pathol. 1999 Jan;30(1):54-8. doi: 10.1016/s0046-8177(99)90300-3.

Abstract

Microsporidia are emerging as opportunistic pathogens in patients with acquired immunodeficiency syndrome (AIDS). Enterocytozoon bieneusi is the most commonly reported microsporidium that is detected in gastrointestinal specimens. This report describes an in situ hybridization technique with a 30-base specific synthetic DNA probe for detection of E bieneusi by light microscopy. Formalin-fixed paraffin-embedded duodenal biopsy specimens from three patients with AIDS, chronic diarrhea, and E bieneusi infection confirmed by electron microscopy were used in this study. Light microscopic examination after colorimetric detection allowed the identification of different stages of the pathogen's life cycle in the cytoplasm of enterocytes. No cross-reactivity was noted between the probe and human DNA. Our study underscores the applicability of a synthetic-labeled oligonucleotide for the detection and identification of E bieneusi in clinical samples.

摘要

微孢子虫正成为获得性免疫缺陷综合征(AIDS)患者的机会性病原体。肠脑炎微孢子虫是在胃肠道标本中检测到的最常报告的微孢子虫。本报告描述了一种使用30个碱基的特异性合成DNA探针通过光学显微镜检测肠脑炎微孢子虫的原位杂交技术。本研究使用了来自三名AIDS患者、患有慢性腹泻且经电子显微镜确诊为肠脑炎微孢子虫感染的福尔马林固定石蜡包埋十二指肠活检标本。比色检测后的光学显微镜检查能够识别病原体在肠上皮细胞胞质中不同的生命周期阶段。未观察到探针与人类DNA之间的交叉反应。我们的研究强调了合成标记寡核苷酸在临床样本中检测和鉴定肠脑炎微孢子虫的适用性。

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