SURVIVAL OF MICROSPORIDIA AFTER EXPOSURE TO DISINFECTANTS AND ENVIRONMENTAL CONDI

接触消毒剂和环境条件后微孢子虫的存活率

• 批准号: 6289462
• 负责人: DANIEL P. FEDORKO
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6289462
• 关键词: Microsporidia; alcohols; chlorine; disinfectants; microorganism culture; microorganism genetics; microorganism growth; quaternary ammonium compound; stainings; temperature

Microsporidia are ubiquitous parasites causing infections in insects, fish, and mammals. Recently microsporidia have been demonstrated to infect humans. These organisms cause ophthalmic and gastrointestinal infections, primarily in patients with acquired immune deficiency syndrome. Several genera of human pathogens have been cultivated in cell culture. Currently, there are no data regarding the ability of the human pathogens Encephalitozoon intestinalis and Encephalitozoon hellem to survive under various environmental conditions. Also, there are no reports regarding the effects of disinfectants on spores of these two species. The survival of microsporidial spores after exposure to disinfectants such as chlorine, alcohol, and quaternary ammonium compounds, and environmental conditions such as elevated temperature and dessication will be studied. Cultivation of microsporidia in the shell vial system using various cell lines has been investigated and microsporidia appear to replicate in both fibroblast and epithelial cell lines. Staining methods are being evaluated to detect microsporidial inclusions in infected cell culture after exposure to disinfectants and environmental conditions. A monoclonal antibody produced by Dr. Theodore Nash (LPD, NIAID) has replaced giemsa staining for detection of infected cells. Although the monoclonal antibody makes detection of infected cells easier, the assay is still cumbersome and tedious. In an effort to replace staining of infected cell cultures, a method for quantitation of microsporidial DNA has been developed. This assay utilizes polymerase chain reaction and a europium-labeled DNA probe and is semi-quantitative.

微孢子虫是一种普遍存在的寄生虫，可引起昆虫、鱼类和哺乳动物的感染。最近微孢子虫已被证明可以感染人类。这些微生物引起眼部和胃肠道感染，主要发生在获得性免疫缺陷综合征患者中。人类病原体的几个属已经在细胞培养物中培养。目前，还没有关于人类病原体脑胞虫和脑胞虫在各种环境条件下生存能力的数据。此外，也没有关于消毒剂对这两个物种孢子影响的报告。将研究暴露于氯、酒精和季铵化合物等消毒剂以及高温和干燥等环境条件后微孢子虫孢子的存活情况。已经研究了使用各种细胞系在壳瓶系统中培养微孢子虫，并且微孢子虫似乎在成纤维细胞和上皮细胞系中复制。正在评估染色方法，以检测暴露于消毒剂和环境条件后感染细胞培养物中的微孢子虫内含物。西奥多纳什博士（LPD，NIAID）生产的单克隆抗体已取代吉姆萨染色用于检测感染细胞。虽然单克隆抗体使感染细胞的检测更容易，但该测定仍然繁琐和乏味。为了取代感染细胞培养物的染色，已经开发了一种用于定量微孢子虫DNA的方法。该检测采用聚合酶链反应和铕标记的DNA探针，是半定量的。