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

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

Author information

1
INSERM U1019, Lille, France CNRS UMR 8204, Lille, France Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Biology & Diversity of Emerging Eukaryotic Pathogens (BDEEP) Laboratory, Université Lille-Nord-de-France, Lille, France magali.chabe@univ-lille2.fr.
2
INSERM U1019, Lille, France CNRS UMR 8204, Lille, France Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Biology & Diversity of Emerging Eukaryotic Pathogens (BDEEP) Laboratory, Université Lille-Nord-de-France, Lille, France Health and Environment Microbiology Laboratory, Azm Center for Research in Biotechnology and Its Applications, Doctoral School of Sciences and Technology, Lebanese University, Tripoli, Lebanon.
3
INSERM U1019, Lille, France CNRS UMR 8204, Lille, France Institut Pasteur de Lille, Center for Infection and Immunity of Lille, Lille, France Biology & Diversity of Emerging Eukaryotic Pathogens (BDEEP) Laboratory, Université Lille-Nord-de-France, Lille, France.
4
GENES DIFFUSION, Douai, France PEGASE-Biosciences, Lille, France.

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

PMID:
24965947
DOI:
10.1093/mmy/myu032
[Indexed for MEDLINE]

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