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. 2020 Jun;27(6):1423-1427.
doi: 10.1016/j.sjbs.2020.04.032. Epub 2020 Apr 23.

Pneumocystis Jirovecii detection and comparison of multiple diagnostic methods with quantitative real-time PCR in patients with respiratory symptoms

Mohammad Y Alshahrani  1 , Mohammed Alfaifi  1 , Irfan Ahmad  1 , Ali Gaithan Alkhathami  1 , Abdulrahim Refdan Hakami  1 , Hafiz Ahmad  2 , Osama M Alshehri  3 , Megh Singh Dhakad  4
Affiliations

Pneumocystis Jirovecii detection and comparison of multiple diagnostic methods with quantitative real-time PCR in patients with respiratory symptoms

Mohammad Y Alshahrani et al. Saudi J Biol Sci. 2020 Jun.

Abstract

Pneumocystis jirovecii (PCP) remains a significant cause of mortality and morbidity in patients with respiratory infections. Accurate diagnosis of PCP is still a diagnostic challenge. Hence, the main objectives were to study the incidence of Pneumocystis Jirovecii pneumonia infection among respiratory problems patients and to compare the real-time quantitative PCR technique with various diagnostic methodologies. Patients who have respiratory symptoms of PCP like breathlessness, cough, and fever were enrolled. Bronchoalveolar lavage (BAL) samples were collected and homogenized, and then smears were prepared for examination by Gomorimethanamine silver staining (GMSS), Immunofluorescent staining (IFAT), Toludine blue O (TBO), and Giemsa staining. Further, RT-PCR was also performed for the detection of PCP. The mean patients' age was 52 (SD ± 16) years. 41% were female, and 59% of the patients were male. Weight loss (80%), fever (92%), cough (100%), and dyspnea (76%) were the most common complaints. Twenty-eight patients have been diagnosed with pulmonary infiltrates using chest X-ray. Out of 100 patients, 35% were positive for PCP. The organism was detected using IFAT in all the 35 specimens, 15 of 35 (42.86%) by GMSS, 8 of 35 (17.6%) by Giemsa stain, and 1 of 35 (2.8%) was detected by TBO stains. RT-PCR showed that 39 patients was found to be positive for PCP. Thirty-five of these 39 patients had a positive IFAT (89.74%); the IFAT was negative or undefined in 4 samples. All 39 patients (100%) had signs and symptoms for PCP. Our results suggest that RT-PCR is still the most highly sensitive method for Pneumocystis Jirovecii detection. In poor resource settings where RT-PCR and IFAT is not available, diagnosis of Pneumocystis jirovecii pneumonia remains a complicated issue. In settings where RT-PCR & IFAT are not available, GMSS staining may be the next best choice to detect PCP.

Keywords: Diagnostic methods; GMSS staining; PCR; Pneumocystis Jirovecii; Respiratory tract infection.

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Figures

Fig. 1
Fig. 1
Comparison of different techniques among respiratory PCP patients.
Fig. 2
Fig. 2
Real time PCR amplification for Pneumocystisjiroveci positive patients.
Fig. 3
Fig. 3
Pneumocysiisjiroveci stained with IFAT seen in BAL sample.
Fig. 4
Fig. 4
Pneumocystisjiroveci stained with GMSS in BAL sample.
See this image and copyright information in PMC

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Further Reading

    1. Philippe M., Hauser J.B., Cornelia L.F., Christian G., Marta F., Michael L., Hitesh P., Victoria M., Barbara A., Martin H., Sarah A.F., Xiaohui C., Flora L., Gillian M., David S.P., David W.D. Multicenter, prospective clinical evaluation of respiratory samples from subjects at risk for pneumocystis jiroveciiinfection by use of a commercial real-time PCR assay. J. Clin. Microbiol. 2011;49(5):1872–1878. - PMC - PubMed
    1. Rodney C., Arcenas J.R., Uhl S.P., Buckwalter A.H., Limper D.C., Glenn D.R., Nancy L.W. A real-time polymerase chain reaction assay for detection of Pneumocystis from bronchoalveolar lavage fluid. Diag. Microbiol. Infect. Dis. 2006;54(3):169–175. - PubMed
    1. Florence R.G., Sorya B., Matthieu R., Pierre T., Stephane J., Olivier D., Sylviane C., Yves L.T., Jean P.G. Diagnosis of Pneumocystis jiroveciipneumonia in immunocompromised patients by realtime PCR: a 4-year prospective study. J. Clin. Microbiol. 2014;52:3370–3376. - PMC - PubMed

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