Development of Specific Sequence-Characterized Amplified Region Markers for Detecting Histoplasma capsulatum in Clinical and Environmental Samples
Por:
De Leon, MGF, Lopez, GA, Taylor, ML, Altamirano, GA, Reyes-Montes, MD
Publicada:
1 mar 2012
Categoría:
Microbiology (medical)
Resumen:
Sequence-characterized amplified region (SCAR) markers, generated by randomly amplified polymorphic DNA (RAPD)-PCR, were developed to detect Histoplasma capsulatum selectively in clinical and environmental samples. A 1,200-bp RAPD-PCR-specific band produced with the 1281-1283 primers was cloned, sequenced, and used to design two SCAR markers, 1281-1283(220) and 1281-1283(230). The specificity of these markers was confirmed by Southern hybridization. To evaluate the relevance of the SCAR markers for the diagnosis of histoplasmosis, another molecular marker (M antigen probe) was used for comparison. To validate 1281-1283(220) and 1281-1283(230) as new tools for the identification of H. capsulatum, the specificity and sensitivity of these markers were assessed for the detection of the pathogen in 36 clinical (17 humans, as well as 9 experimentally and 10 naturally infected nonhuman mammals) and 20 environmental (10 contaminated soil and 10 guano) samples. Although the two SCAR markers and
Filiaciones:
De Leon, MGF:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol Parasitol, Mexico City 04510, DF, Mexico
Lopez, GA:
Univ Nacl Autonoma Mexico, Fac Med, Dept Fisiol, Mexico City 04510, DF, Mexico
Taylor, ML:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol Parasitol, Mexico City 04510, DF, Mexico
Altamirano, GA:
Hosp Juarez Mexico, Mexico City, DF, Mexico
Reyes-Montes, MD:
Univ Nacl Autonoma Mexico, Fac Med, Dept Microbiol Parasitol, Mexico City 04510, DF, Mexico
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