Usefulness of molecular markers in the diagnosis of occupational and recreational histoplasmosis outbreaks |
| |
Authors: | María Guadalupe Frías-De-León José Antonio Ramírez-Bárcenas Gabriela Rodríguez-Arellanes Oscar Velasco-Castrejón Maria Lucia Taylor María del Rocío Reyes-Montes |
| |
Institution: | 1.División de Investigación,Hospital Juárez de México,Ciudad de Mexico, Cd.Mx.,Mexico;2.Departamento de Microbiología y Parasitología, Facultad de Medicina,Universidad Nacional Autónoma de México (UNAM),Ciudad de México, Cd.Mx.,Mexico;3.Hospital General de México,Ciudad de Mexico Cd.Mx.,Mexico |
| |
Abstract: | Histoplasmosis is considered the most important systemic mycosis in Mexico, and its diagnosis requires fast and reliable methodologies. The present study evaluated the usefulness of PCR using Hcp100 and 1281–1283(220) molecular markers in detecting Histoplasma capsulatum in occupational and recreational outbreaks. Seven clinical serum samples of infected individuals from three different histoplasmosis outbreaks were processed by enzyme-linked immunosorbent assay (ELISA) to titre anti-H. capsulatum antibodies and to extract DNA. Fourteen environmental samples were also processed for H. capsulatum isolation and DNA extraction. Both clinical and environmental DNA samples were analysed by PCR with Hcp100 and 1281–1283(220) markers. Antibodies to H. capsulatum were detected by ELISA in all serum samples using specific antigens, and in six of these samples, the PCR products of both molecular markers were amplified. Four environmental samples amplified one of the two markers, but only one sample amplified both markers and an isolate of H. capsulatum was cultured from this sample. All PCR products were sequenced, and the sequences for each marker were analysed using the Basic Local Alignment Search Tool (BLASTn), which revealed 95–98 and 98–100 % similarities with the reference sequences deposited in the GenBank for Hcp100 and 1281–1283(220), respectively. Both molecular markers proved to be useful in studying histoplasmosis outbreaks because they are matched for pathogen detection in either clinical or environmental samples. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|