Departamento de Microbiologia e Imunologia, Instituto de Biociências de Botucatu, Universidade Estadual Paulista (UNESP), Botucatu, Brazil.
Med Mycol. 2013 Jan;51(1):83-92. doi: 10.3109/13693786.2012.698444. Epub 2012 Jul 5.
Taking into account that paracoccidioidomycosis infection occurs by inhalation of the asexual conidia produced by Paracoccidioides spp. in its saprobic phase, this work presents the collection of aerosol samples as an option for environmental detection of this pathogen, by positioning a cyclonic air sampler at the entrance of armadillo burrows. Methods included direct culture, extinction technique culture and Nested PCR of the rRNA coding sequence, comprising the ITS1-5.8S-ITS2 region. In addition, we evaluated one armadillo (Dasypus novemcinctus) as a positive control for the studied area. Although the pathogen could not be isolated by the culturing strategies, the aerosol sampling associated with molecular detection through Nested PCR proved the best method for discovering Paracoccidioides spp. in the environment. Most of the ITS sequences obtained in this investigation proved to be highly similar with the homologous sequences of Paracoccidioides lutzii from the GenBank database, suggesting that this Paracoccidioides species may not be exclusive to mid-western Brazil as proposed so far.
考虑到副球孢子菌病感染是通过吸入副球孢子菌属在腐生阶段产生的无性分生孢子而发生的，因此本工作提出了收集气载样本作为该病原体环境检测的一种选择，方法是在犰狳洞穴入口处放置旋风空气采样器。方法包括直接培养、灭活该病原体培养和 rRNA 编码序列的嵌套 PCR，包含 ITS1-5.8S-ITS2 区域。此外，我们还评估了一只犰狳（Dasypus novemcinctus）作为研究区域的阳性对照。尽管通过培养策略无法分离出病原体，但与分子检测相关的气溶胶采样通过嵌套 PCR 证明是发现环境中副球孢子菌属的最佳方法。在本研究中获得的大多数 ITS 序列与 GenBank 数据库中副球孢子菌属 lutzii 的同源序列高度相似，这表明该副球孢子菌种可能不像迄今为止所提出的那样仅限于巴西中西部。
