A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels

• DOI: 10.3389/fmicb.2021.787554
• Publication Date: 2022-01-11T05:22:17Z
• Authors: Susana Ruiz-Ruiz; Carolina A. Ponce; Nicole Pesantes; Rebeca Bustamante; Gianna Gatti; Viviana San Martin; Mireya Gutierrez; Pamela Bórquez; Sergio L. Vargas; Fabien Magne; Enrique J. Calderón; Vicente Pérez-Brocal; Andrés Moya
• Keywords: Primer (cosmetics)

Here we report a new real-time PCR assay using SYBR Green which provides higher sensitivity for the specific detection of low levels Pneumocystis jirovecii . To do so, two primer sets were designed, targeting family genes that code most abundant surface protein spp., namely major glycoproteins (Msg), and mitochondrial large subunit rRNA (mtLSUrRNA) multicopy gene, simultaneously detecting regions. methods are instrumental in these levels; however, current nested-PCR time-consuming complex. validate our Msg-A/mtLSUrRNA protocol, compared it with based on (mtLSUrRNA), one main targets used to detect this pathogen. All samples identified as positive by method found also detected P. three nasal aspirate negative both rounds nested-PCR. Furthermore, read results comparison some no amplification, or feeble band first round, correlated Ct values PCR. This finding demonstrates ability single-round protocol levels. valuable alternative detection, is rapid sensitive.