RT Journal Article
T1 A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels.
A1 Ruiz-Ruiz, Susana
A1 Ponce, Carolina A
A1 Pesantes, Nicole
A1 Bustamante, Rebeca
A1 Gatti, Gianna
A1 San Martin, Viviana
A1 Gutierrez, Mireya
A1 Borquez, Pamela
A1 Vargas, Sergio L
A1 Magne, Fabien
A1 Calderon, Enrique J
A1 Perez-Brocal, Vicente
A1 Moya, Andres
K1 low P. jirovecii loads
K1 major surface glycoproteins
K1 mitochondrial large subunit rRNA
K1 real-time PCR
K1 simultaneous detection
AB Here we report a new real-time PCR assay using SYBR Green which provides higher sensitivity for the specific detection of low levels of Pneumocystis jirovecii. To do so, two primer sets were designed, targeting the family of genes that code for the most abundant surface protein of Pneumocystis spp., namely the major surface glycoproteins (Msg), and the mitochondrial large subunit rRNA (mtLSUrRNA) multicopy gene, simultaneously detecting two regions. PCR methods are instrumental in detecting these low levels; however, current nested-PCR methods are time-consuming and complex. To validate our new real-time Msg-A/mtLSUrRNA PCR protocol, we compared it with nested-PCR based on the detection of Pneumocystis mitochondrial large subunit rRNA (mtLSUrRNA), one of the main targets used to detect this pathogen. All samples identified as positive by the nested-PCR method were found positive using our new real-time PCR protocol, which also detected P. jirovecii in three nasal aspirate samples that were negative for both rounds of nested-PCR. Furthermore, we read both rounds of the nested-PCR results for comparison and found that some samples with no PCR amplification, or with a feeble band in the first round, correlated with higher Ct values in our real-time Msg-A/mtLSUrRNA PCR. This finding demonstrates the ability of this new single-round protocol to detect low Pneumocystis levels. This new assay provides a valuable alternative for P. jirovecii detection, as it is both rapid and sensitive.
PB Frontiers Research Foundation
SN 1664-302X
YR 2022
FD 2022-01-11
LK http://hdl.handle.net/10668/20615
UL http://hdl.handle.net/10668/20615
LA en
NO Ruiz-Ruiz S, Ponce CA, Pesantes N, Bustamante R, Gatti G, San Martin V, et al. A Real-Time PCR Assay for Detection of Low Pneumocystis jirovecii Levels. Front Microbiol. 2022 Jan 11;12:787554.
DS RISalud
RD Jul 30, 2025