Rapid detection and identification of fungal pathogens by rolling circle amplification (RCA) using Fonsecaea as a model

摘要: We aimed to describe a rapid and sensitive assay for identification of pathogenic fungi without sequencing. The method rolling circle amplification (RCA) is presented with species Fonsecaea, agents human chromoblastomycosis, as model. internal transcribed spacer (ITS) rDNA region 103 Fonsecaea strains was sequenced aligned in view designing three specific padlock probes be used the detection single nucleotide polymorphisms species. 38 included testing specificity RCA comprised 17 isolates pedrosoi, 13 monophora eight nubica. successfully amplified DNA target at level species, while no cross reactivity observed. product visualised on 1% agarose gel verify probe–template binding. Amounts reagents were minimised avoid generation false-positive results. simplicity, sensitivity, robustness low costs provide distinct position among isothermal techniques diagnostics very practical method.