Abstract
Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods. The OTsn-PCR assay was positive for all clinical samples, and the detection limit was better or equivalent to the other nested or semi-nested PCR methods for P. brasiliensis detection. The OTsn-PCR assay described in this paper has a detection limit similar to other reactions for the molecular detection of P. brasiliensis, but this approach is faster and less prone to contamination than other conventional nested or semi-nested PCR assays.
Highlights
We report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis
Using a serial dilution of purified Deoxyribonucleic acid (DNA) from P. brasiliensis, the OTsn-PCR assay described in this study was able to detect up to 2.5pg of P. brasiliensis DNA (Figure 1A) without cross-reactivity with other fungi
The limit of detection for the OTsn-PCR assay was comparable with that of the protocol developed by Koishi et al.[5] (Figure 1B) and was more sensitive than the protocols in
Summary
We report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis. Several new molecular methods have been developed for the detection of P. brasiliensis, including polymerase chain reaction (PCR) assays[3,4].
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