title: An improved single-round PCR leads to rapid and highly sensitive detection of Pneumocystis spp.
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.
authors: 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.
authors: Magali
authors: M
authors: Chabé
authors: 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.
authors: Sara
authors: S
authors: Khalife
authors: 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.
authors: Nausicaa
authors: N
authors: Gantois
authors: GENES DIFFUSION, Douai, France PEGASE-Biosciences, Lille, France.
authors: Gaël
authors: G
authors: Even
authors: GENES DIFFUSION, Douai, France PEGASE-Biosciences, Lille, France.
authors: Christophe
authors: C
authors: Audebert
mesh_terms: D000818:Animals; D001483:Base Sequence; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D006801:Humans; D057230:Limit of Detection; D008969:Molecular Sequence Data; D009960:Oropharynx / Q000382:microbiology; D011010:Pneumocystis / Q000235:genetics* / Q000302:isolation & purification; D016720:Pneumocystis Infections / Q000175:diagnosis / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
keywords: PneumoDB; Pneumocystis spp.; mtLSUrRNA; single-round PCR