An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection.
BACKGROUND
Herein, we report a one-tube, semi-nested-polymerase chain reaction (OTsn-PCR) assay for the detection of Paracoccidioides brasiliensis.


METHODS
We developed the OTsn-PCR assay for the detection of P. brasiliensis in clinical specimens and compared it with other PCR methods.


RESULTS
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.


CONCLUSIONS
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.
Amanda de Faveri
Ade F
Pitz
Andrea Cristine
AC
Koishi
Eliandro Reis
ER
Tavares
Fábio Goulart de
FG
Andrade
Eduardo Alexandre
EA
Loth
Rinaldo Ferreira
RF
Gandra
Emerson José
EJ
Venancio
D004271:DNA, Fungal / Q000302:isolation & purification; D006801:Humans; D010228:Paracoccidioides / Q000235:genetics / Q000302:isolation & purification*; D010229:Paracoccidioidomycosis / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity
D004271:DNA, Fungal