Performance of the RealStar^®Pneumocystis jirovecii PCR kit for the diagnosis of Pneumocystis pneumonia

Abstract

Background Pneumonia due to Pneumocystis jirovecii (PCP) is a frequent infection in HIV‐positive and also in immunocompromised HIV‐negative patients. PCR analysis of pulmonary samples has become an essential element in PCP laboratory diagnosis. Currently, many commercially PCR‐based tests are available for P jirovecii detection and need to be evaluated. Objectives We evaluated the performance of the RealStar® P jirovecii PCR kit for PCP diagnosis. Methods We performed the RealStar® P jirovecii PCR and an in‐house PCR in 219 pulmonary samples. We then assessed the performance of the RealStar® P jirovecii PCR kit by classifying patients in proven, probable, possible PCP or no final diagnosis, on the basis of the clinical and radiological signs and direct examination of bronchoalveolar lavage samples. Results The results showed excellent concordance (96.8%) with another in‐house PCR, previously used in the laboratory. The available clinical data allowed classifying 219 patients as having proven PCP (n = 6), probable PCP (n = 27), possible PCP (n = 29) and no final diagnosis of PCP (n = 157). The RealStar® P jirovecii PCR kit performed well with samples from patients with proven and probable PCP, as indicated by the detection of P jirovecii DNA in all these samples. The percentage of positive samples in the possible PCP category was 75.9%. In patients with no final diagnosis of PCP, P jirovecii DNA was detected in 13.4% of samples, indicating colonisation by this pathogen. Conclusions The RealStar® P jirovecii PCR kit shows excellent performance for PCP diagnosis.

Quick versus Quantitative: Evaluation of Two Commercial Real-Time PCR Assays for the Detection of Pneumocystis jirovecii from Bronchoalveolar Lavage Fluids

“…Recent studies have demonstrated the superior sensitivity of the DiaSorin PJP assay when compared to a direct immunofluorescence assay ( 28 ) and its clinical usefulness for the diagnosis and management of PJP in correlation with clinical data and pathology testing ( 27 ). The RealStar assay has shown excellent concordance with several other molecular PJP assays and has been reported to detect low burdens of P. jirovecii in respiratory specimens ( 24 – 26 ).…”

“…Both in-house and commercial real-time PCR assays for the detection of P. jirovecii using different gene targets have been developed ( 20 , 22 , 23 ). Among P. jirovecii real-time PCR assays on the market today are the altona RealStar Pneumocystis jirovecii PCR kit 1.0 ( 24 – 26 ) and the DiaSorin P. jirovecii real-time assay ( 27 , 28 ). Both assays target the multicopy mitochondrial large-subunit rRNA ( mtLSU ) gene ( 26 , 28 ).…”

“…Among P. jirovecii real-time PCR assays on the market today are the altona RealStar Pneumocystis jirovecii PCR kit 1.0 ( 24 – 26 ) and the DiaSorin P. jirovecii real-time assay ( 27 , 28 ). Both assays target the multicopy mitochondrial large-subunit rRNA ( mtLSU ) gene ( 26 , 28 ). The RealStar PJP assay provides quantitative results from extracted nucleic acids based on a standard curve, whereas the DiaSorin assay offers the convenience of real-time PCR detection of P. jirovecii directly from respiratory specimens without prior nucleic acid extraction.…”

Relationship between clinical features and droplet digital PCR copy number in non-HIV patients with pneumocystis pneumonia

“…In recent years, molecular detection methods have gradually been developed to detect Pj . Polymerase chain reaction (PCR) assays, which mainly target the mitochondrial large subunit rRNA gene and the multicopy major surface glycoprotein gene family, make the detection of Pj more sensitive than microscopic detection [ 7 , 8 ]. However, no commercial PCR kits have been approved for clinical use in China, and their sensitivity is unsatisfactory [ 9 ].…”

Multilocus Genotyping of Pneumocystis jirovecii from Deceased Cuban AIDS Patients Using Formalin-Fixed and Paraffin-Embedded Tissues

“…In this sense, the samples obtained by bronchoscopy provide a better score of positive detection. However, collecting this type of sample requires invasive medical management, therefore this procedure is not usually performed in many countries to identify the pathogen [1,7]. In view of this limitation, formalin-fixed and paraffin-embedded tissues (FFPE) could be alternative sources of samples for molecular studies [8,9].…”