Role of Cytology and Polymerase Chain Reaction Based Detection of Pneumocystis jirovecii Infection in Bronchoalveolar Lavage Fluid

Gupta, Rashmi; Iyer, Venkateswaran K.; Mirdha, Bijay Ranjan; Guleria, Randeep; Kumar, Lalit; Agarwal, Sanjay Kumar

doi:10.1159/000325038

Cited by 4 publications

(2 citation statements)

References 26 publications

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“…This approach is intended to facilitate differentiation between colonization and infection [9]. Nevertheless, no clear answer as to how to distinguish these two situations by PCR methods has been given yet [10]. The PCR assay, widely used in diagnosing PCP from BAL or lung samples, is also a promising technology for respiratory samples obtained by non-invasive methods such as induced sputum, nasopharyngeal aspirate, or oral washing specimens [1, 8].…”

Section: Introductionmentioning

confidence: 99%

Clinical immunology Pneumocystis pneumonia in children – the relevance of chemoprophylaxis in different groups of immunocompromised and immunocompetent paediatric patients

Zając‐Spychała¹,

Gowin²,

Fichna³

et al. 2015

cejoi

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IntroductionPneumocystis jirovecii is an opportunistic pathogen causing pneumocystis pneumonia (PCP), a life-threatening infection, in immunocompromised patients. In this study, retrospective analysis of the presence of P. jirovecii DNA in different samples collected from children with suspected PCP was carried out.Material and methodsThree hundred and six specimens [152 bronchoalveolar lavage (BAL) specimens, 80 blood specimens, 18 bronchial secretions (BS), 34 induced sputum samples, 10 endotracheal aspirates (ETA), and 12 other type samples] obtained from patients with suspected PCP were examined by real-time PCR.ResultsForty (13.1%) patients were positive for P. jirovecii: 4 (7.7%) patients with malignancies, 3 (6.8%) transplant recipients, 15 (23.1%) other immunocompromised patients, and 18 (12.4%) immunocompetent patients. Pneumocystis jirovecii DNA was detected in 20.4% of BAL specimens, 11.1% of BS samples, 10% of ETA sample, 8.8% of induced sputum samples, and in 3.7% of blood samples. Comparing the frequency of the presence of P. jirovecii DNA between the group of children treated with PCP chemoprophylaxis (malignancy patients and transplant recipients) and a group of children not receiving this prophylaxis (other immunocompromised and immunocompetent children), we found that the occurrence of PCP was twice as high in the latter group of children (7.3% and 15.7%, respectively).ConclusionsRespiratory samples, such as BS, BAL, or ETA specimens, are the material of choice for the diagnosis of PCP. Due to high incidence of PCP in certain groups of immunocompetent and immunocompromised patients, besides cancer patients and transplant recipients, consideration of PCP prophylaxis is required in these groups as well.

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Section: Introductionmentioning

confidence: 99%

Clinical immunology Pneumocystis pneumonia in children – the relevance of chemoprophylaxis in different groups of immunocompromised and immunocompetent paediatric patients

Zając‐Spychała¹,

Gowin²,

Fichna³

et al. 2015

cejoi

View full text Add to dashboard Cite

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“…PCR is more sensitive, and both single-round and nested assays have been developed (Su et al, 2008;Gupta et al, 2010;Jiancheng et al, 2009;Jarboui et al, 2010). These increase sensitivity but are thought to reduce clinical specificity (Mekinian et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Rising incidence of Pneumocystis jirovecii pneumonia suggests iatrogenic exposure of immune-compromised patients may be becoming a significant problem

Coyle

McCaughey

Nager

et al. 2012

Journal of Medical Microbiology

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Against a background of point-source outbreaks of Pneumocystis pneumonia (PCP) in renal transplant units in Europe, we undertook a retrospective 3 year observational review of PCP in Northern Ireland. This showed an unexpected increase in incidence, with a mortality rate of 30 %. Fifty-one cases were confirmed compared to 10 cases confirmed in the preceding 7 years. Where undiagnosed HIV infection had previously been the main risk factor for PCP, this was now equally matched by chemotherapy for haematological and non-haematological malignancy and immune suppression for a range of autoimmune conditions. Congenital immunodeficiency and transplantation were less common predisposing factors, but renal grafts also showed a rising incidence. Asymptomatic carriage was uncommon. At presentation both upper and lower respiratory samples were of equal use in establishing the diagnosis, and treatment resulted in rapid clearance. These data suggest the need for considering PCP in at-risk patients, reviewing its mode of acquisition and whether iatrogenic colonization is a treatable pre-condition.

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Axenic Long-Term Cultivation of Pneumocystis jirovecii

Riebold,

Mahnkopf,

Wicht

et al. 2023

JoF

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Pneumocystis jirovecii, a fungus causing severe Pneumocystis pneumonia (PCP) in humans, has long been described as non-culturable. Only isolated short-term experiments with P. jirovecii and a small number of experiments involving animal-derived Pneumocystis species have been published to date. However, P. jirovecii culture conditions may differ significantly from those of animal-derived Pneumocystis, as there are major genotypic and phenotypic differences between them. Establishing a well-performing P. jirovecii cultivation is crucial to understanding PCP and its pathophysiological processes. The aim of this study, therefore, was to develop an axenic culture for Pneumocystis jirovecii. To identify promising approaches for cultivation, a literature survey encompassing animal-derived Pneumocystis cultures was carried out. The variables identified, such as incubation time, pH value, vitamins, amino acids, and other components, were trialed and adjusted to find the optimum conditions for P. jirovecii culture. This allowed us to develop a medium that produced a 42.6-fold increase in P. jirovecii qPCR copy numbers after a 48-day culture. Growth was confirmed microscopically by the increasing number and size of actively growing Pneumocystis clusters in the final medium, DMEM-O3. P. jirovecii doubling time was 8.9 days (range 6.9 to 13.6 days). In conclusion, we successfully cultivated P. jirovecii under optimized cell-free conditions in a 70-day long-term culture for the first time. However, further optimization of the culture conditions for this slow grower is indispensable.

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Role of Cytology and Polymerase Chain Reaction Based Detection of Pneumocystis jirovecii Infection in Bronchoalveolar Lavage Fluid

Cited by 4 publications

References 26 publications

Clinical immunology Pneumocystis pneumonia in children – the relevance of chemoprophylaxis in different groups of immunocompromised and immunocompetent paediatric patients

Clinical immunology Pneumocystis pneumonia in children – the relevance of chemoprophylaxis in different groups of immunocompromised and immunocompetent paediatric patients

Rising incidence of Pneumocystis jirovecii pneumonia suggests iatrogenic exposure of immune-compromised patients may be becoming a significant problem

Axenic Long-Term Cultivation of Pneumocystis jirovecii

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