An optimized one-tube, semi-nested PCR assay for Paracoccidioides brasiliensis detection

Rev Soc Bras Med Trop. 2013 Nov-Dec;46(6):783-5. doi: 10.1590/0037-8682-1625-2013.

Abstract

Introduction: 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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Fungal / isolation & purification
  • Humans
  • Paracoccidioides / genetics
  • Paracoccidioides / isolation & purification*
  • Paracoccidioidomycosis / diagnosis*
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • DNA, Fungal