An improved single-round PCR leads to rapid and highly sensitive detection of Pneumocystis spp

Med Mycol. 2014 Nov;52(8):841-6. doi: 10.1093/mmy/myu032. Epub 2014 Jun 25.

Abstract

In order to standardize a polymerase chain reaction (PCR)-based method of Pneumocystis detection, we describe the development of an improved PCR method that targets the Pneumocystis mtLSUrRNA gene. Design of a new primer pair and PCR program with suitable parameters and optimization resulted in a simpler and faster single-round amplification assay. The sensitivity of the novel Pneumocystis genus-specific PCR proved comparable to the reference nested PCR. The improvement that this new PCR assay offers in the detection and epidemiological studies of Pneumocystis spp. infection in research laboratories is discussed.

Keywords: PneumoDB; Pneumocystis spp.; mtLSUrRNA; single-round PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Bronchoalveolar Lavage Fluid / microbiology
  • Humans
  • Limit of Detection
  • Molecular Sequence Data
  • Oropharynx / microbiology
  • Pneumocystis / genetics*
  • Pneumocystis / isolation & purification
  • Pneumocystis Infections / diagnosis
  • Pneumocystis Infections / microbiology
  • Polymerase Chain Reaction / methods*
  • Rats
  • Sensitivity and Specificity
  • Sequence Alignment