title: Usefulness of molecular markers in the diagnosis of occupational and recreational histoplasmosis outbreaks.
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.
authors.affiliation: División de Investigación, Hospital Juárez de México, Av. Instituto Politécnico Nacional 5160, Col. Magdalena de las Salinas, 07760, Ciudad de Mexico, Cd.Mx., Mexico.
authors.forename: María Guadalupe
authors.initials: MG
authors.lastname: Frías-De-León
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria No. 3000, Ciudad de México, Cd.Mx., 04510, Mexico.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Ramírez-Bárcenas
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria No. 3000, Ciudad de México, Cd.Mx., 04510, Mexico.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Rodríguez-Arellanes
authors.affiliation: Hospital General de México, Dr. Balmis No. 148, Col. Doctores, 06726, Ciudad de Mexico Cd.Mx., Mexico.
authors.forename: Oscar
authors.initials: O
authors.lastname: Velasco-Castrejón
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria No. 3000, Ciudad de México, Cd.Mx., 04510, Mexico.
authors.forename: Maria Lucia
authors.initials: ML
authors.lastname: Taylor
authors.affiliation: Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Ciudad Universitaria No. 3000, Ciudad de México, Cd.Mx., 04510, Mexico. remoa@unam.mx.
authors.forename: María Del Rocío
authors.initials: MDR
authors.lastname: Reyes-Montes
mesh_terms: D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood*; D004271:DNA, Fungal / Q000235:genetics*; D004196:Disease Outbreaks; D004797:Enzyme-Linked Immunosorbent Assay; D005819:Genetic Markers; D006658:Histoplasma / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006660:Histoplasmosis / Q000175:diagnosis* / Q000453:epidemiology* / Q000382:microbiology; D006801:Humans; D008297:Male; D008800:Mexico / Q000453:epidemiology; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D012988:Soil Microbiology
chemical_list: D000908:Antibodies, Fungal; D004271:DNA, Fungal; D005819:Genetic Markers