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Molecular Detection of Pneumocystis jirovecii in the Bronchoalveolar Lavage Samples in Patients with Various Pulmonary Disorders
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Narges Chitsaz    , Ahmad Reza Meamar , Elham Razmjou , Soheila Shafaghi-Sisi , Maryam Alipour , Maryam Sadeghi , Zahra Rampisheh , Zeinab Ghasemi , Rasoul Aliannejad , Alireza Badirzadeh * |
| Department of Parasitology and Mycology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran , badirzadeh.ar@iums.ac.ir |
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Abstract: (557 Views) |
Background: Pneumocystis jirovecii (P. jirovecii) causes Pneumocystis pneumonia (PCP) in people, especially the immunocompromised ones. It is also one of the serious causes of numerous lung problems in affected patients. Since documented data about P. jirovecii is not available in patients with pulmonary infections in Tehran, this study aimed to investigate the molecular epidemiology and parasitology of Pneumocystis to determine the frequency of the organism infection.
Methods: Bronchoalveolar lavage (BAL) samples were collected for 367 patients hospitalized in the lung department of Shariati Hospital in Tehran from July 2022 to July 2023. The samples were analyzed using Giemsa staining and molecular methods. After DNA extraction from samples, Nested polymerase chain reaction (Nested PCR) was employed for the amplification of the 18SrRNA gene and identification of P. jirovecii. The PCR products of Nested PCR were sequenced for final confirmation.
Results: Out of 367 samples, only one sample (0.27%) and 28 samples (6.7%) were found to be positive through parasitology and NestedPCR analysis, respectively. P. jirovecii was detected in seven (25%) and 21 (75%) immunocompromised and immunocompetent patients, respectively. Fever, shortness of breath and dry cough were the most common clinical symptoms among patients with Pneumocystosis. Patients with pulmonary disorders are prone to colonization by pneumocystis, which increases the risk of pneumocystosis and makes them a reservoir for transmission to susceptible people.
Conclusion: It can be concluded that patients with distinct lung disease are prone to colonization by Pneumocystis and, importantly, are at risk of infection. Also, according to the current study, Nested PCR was a suitable method for detecting P. jirovecii organisms because it had a very high sensitivity and specificity.
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Article number: 1 |
| Keywords: Pneumocystis jirovecii, Pneumonia, Colonization, Nested-PCR, Broncho-alveolar lavage (BAL) |
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Full-Text [PDF 652 kb]
(276 Downloads)
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Type of Study: article |
Subject:
Parasitology-Mycology-Entomology
Received: 2023/02/20 | Accepted: 2023/06/27 | Published: 2023/12/31
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1. Meenakshi K, Gowtham, RR, USha, K. Pneumocystis jirovecii Pneumonia: a Revisit to the old Malady. J Clin Diagn Res. 2019; 13(11):1-8. [ DOI:10.7860/JCDR/2019/42636.13318] 2. Hammarström H, Grankvist A, Broman I, Kondori N, Wennerås C, Gisslen M, et al. Serum-based diagnosis of Pneumocystis pneumonia by detection of Pneumocystis jirovecii DNA and 1, 3-β-D-glucan in HIV-infected patients: a retrospective case control study. BMC Infect Dis. 2019; 19(1):1-10. [ DOI:10.1186/s12879-019-4289-4] [ PMID] [ ] 3. Mohebali M, Mirbakhsh M, Keshavarz H. Rapid detection of Pneumocystis carinii in spiratory specimens of rats by calcofluor white staining. Iran J Public Health. 2002.31(3-4):108-110. 4. Dellière S, Gits-Muselli M, Bretagne S, Alanio A. Outbreak-causing fungi: Pneumocystis jirovecii. Mycopathologia. 2020; 185(5):783-800. [ DOI:10.1007/s11046-019-00408-w] [ PMID] 5. Nevez G, Guillaud-Saumur T, Cros P, Papon N, Vallet S, Quinio D, et al., Pneumocystis primary infection in infancy: additional french data and review of the literature. Med Mycol. 2020; 58(2):163-171. [ DOI:10.1093/mmy/myz040] [ PMID] [ ] 6. Fauchier T, Hasseine L, Gari-Toussaint M, Casanova V, Marty PM, Pomares C. Detection of Pneumocystis jirovecii by quantitative PCR to differentiate colonization and pneumonia in immunocompromised HIV-positive and HIV-negative patients. J Clin Microbiol. 2016; 54(6):1487-1495. [ DOI:10.1128/JCM.03174-15] [ PMID] [ ] 7. Vera C, Rueda ZV. Transmission and colonization of Pneumocystis jirovecii. J Fungi. 2021; 7(11):979-995. [ DOI:10.3390/jof7110979] [ PMID] [ ] 8. Lagrou K, Chen S, Masur H, Viscoli C, Decker CF, Pagano L, et al. Pneumocystis jirovecii disease: basis for the revised EORTC/MSGERC invasive fungal disease definitions in individuals without human immunodeficiency virus. Clin Infect Dis. 2021; 72(Supplement_2):114-120. [ DOI:10.1093/cid/ciaa1805] [ PMID] [ ] 9. Karageorgopoulos DE, Qu JM, Korbila IP, Zhu YG, Vasileiou VA, Falagas ME. Accuracy of β-D-glucan for the diagnosis of Pneumocystis jirovecii pneumonia: a meta-analysis. Clin Microbiol Infect. 2013; 19(1):39-49. [ DOI:10.1111/j.1469-0691.2011.03760.x] [ PMID] 10. Pennington K, Wilson J, Limper AH, Escalante P. Positive Pneumocystis jirovecii sputum PCR results with negative bronchoscopic PCR results in suspected Pneumocystis pneumonia. Can Respir J. 2018; 2018:1-5. [ DOI:10.1155/2018/6283935] [ PMID] [ ] 11. Fan LC, Lu HW, Cheng KB, Li HP, Xu JF. Evaluation of PCR in bronchoalveolar lavage fluid for diagnosis of Pneumocystis jirovecii pneumonia: a bivariate meta-analysis and systematic review. PLoS One. 2013; 8(9):e73099. [ DOI:10.1371/journal.pone.0073099] [ PMID] [ ] 12. Khodadadi H, Mirhendi H, Mohebali M, Kordbacheh P, Zarrinfar H, Makimura K. Pneumocystis jirovecii colonization in non-HIV-infected patients based on nested-PCR detection in bronchoalveolar lavage samples. Iran J Public Health. 2013; 42(3):298-305. 13. Gantois N, Lesaffre A, Durand-Joly I, Bautin N, Le Rouzic O, Nseir S, et al. Factors associated with Pneumocystis colonization and circulating genotypes in chronic obstructive pulmonary disease patients with acute exacerbation or at stable state and their homes. Med Mycol. 2022; 60(1): myab070. [ DOI:10.1093/mmy/myab070] [ PMID] 14. Maskell NA, Waine DJ, Lindley A, Pepperell JC, Wakefield AE, Miller RF, et al. Asymptomatic carriage of Pneumocystis jiroveci in subjects undergoing bronchoscopy: a prospective study. Thorax. 2003; 58(7):594-597. [ DOI:10.1136/thorax.58.7.594] [ PMID] [ ] 15. Rozaliyani A, Sjam R, Tugiran M, Adawiyah R, Dwiatmo C, Haryanto B, et al. Detection of Pneumocystis jirovecii by nested PCR in lung cancer patients in a tertiary hospital in Jakarta: A preliminary study. Int J Infect Dis. 2020; 101:386-387. [ DOI:10.1016/j.ijid.2020.09.1014] 16. Flori P, Bellete B, Durand F, Raberin H, Cazorla C, Hafid J, et al. Comparison between real-time PCR, conventional PCR and different staining techniques for diagnosing Pneumocystis jiroveci pneumonia from bronchoalveolar lavage specimens. J Med Microbiol. 2004; 53(7):603-607. [ DOI:10.1099/jmm.0.45528-0] [ PMID] 17. Parvizi P, Ready PD. Nested PCRs and sequencing of nuclear ITS‐rDNA fragments detect three Leishmania species of gerbils in sandflies from Iranian foci of zoonotic cutaneous leishmaniasis. Trop Med Int Health. 2008; 13(9):1159-1171. [ DOI:10.1111/j.1365-3156.2008.02121.x] [ PMID] 18. Morley AA. Digital PCR: A brief history. Biomol Detect Quantif. 2014; 1(1):1-2. [ DOI:10.1016/j.bdq.2014.06.001] [ PMID] [ ] 19. Eisenstein BI. The polymerase chain reaction: a new method of using molecular genetics for medical diagnosis. N Engl J Med. 1990; 322(3):178-183. [ DOI:10.1056/NEJM199001183220307] [ PMID] 20. Baumforth KR, Nelson PN, Digby JE, O'Neil JD, Murray PG. Demystified the polymerase chain reaction. Mol Pathol. 1999; 52(1):1-1. [ DOI:10.1136/mp.52.1.1] [ PMID] [ ] 21. Bateman M, Oladele R, Kolls JK. Diagnosing Pneumocystis jirovecii pneumonia: A review of current methods and novel approaches. Med Mycol. 2020; 58(8):1015-1028. [ DOI:10.1093/mmy/myaa024] [ PMID] [ ] 22. McTaggart LR, Wengenack NL, Richardson SE. Validation of the MycAssay Pneumocystis kit for detection of Pneumocystis jirovecii in bronchoalveolar lavage specimens by comparison to a laboratory standard of direct immunofluorescence microscopy, real-time PCR, or conventional PCR. J Clin Microbiol. 2012; 50(6):1856-1859. [ DOI:10.1128/JCM.05880-11] [ PMID] [ ] 23. Robberts FJ, Liebowitz LD, Chalkley LJ. Polymerase chain reaction detection of Pneumocystis jiroveci: evaluation of 9 assays. Diagn Microbiol Infect Dis. 2007; 58(4):385-392. [ DOI:10.1016/j.diagmicrobio.2007.02.014] [ PMID] 24. Alanio A, Desoubeaux G, Sarfati C, Hamane S, Bergeron A, Azoulay E, et al. Real-time PCR assay-based strategy for differentiation between active Pneumocystis jirovecii pneumonia and colonization in immunocompromised patients. Clin Microbiol Infect. 2011; 17(10):1531-1537. [ DOI:10.1111/j.1469-0691.2010.03400.x] [ PMID] 25. Maillet M, Maubon D, Brion JP, François P, Molina L, Stahl JP, et al. Pneumocystis jirovecii (Pj) quantitative PCR to differentiate Pj pneumonia from Pj colonization in immunocompromised patients. Eur J Clin Microbiol Infect Dis. 2014; 33(3):331-336. [ DOI:10.1007/s10096-013-1960-3] [ PMID] [ ] |
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Ethics code: IR.IUMS.FMD.REC.1400.536
Chitsaz N, Meamar A R, Razmjou E, Shafaghi-Sisi S, Alipour M, Sadeghi M, et al . Molecular Detection of Pneumocystis jirovecii in the Bronchoalveolar Lavage Samples in Patients with Various Pulmonary Disorders. J Ardabil Univ Med Sci 2024; 23 (4) : 1 URL: http://jarums.arums.ac.ir/article-1-2262-en.html
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