scholarly journals Diagnosis of Pneumocystis jirovecii Pneumonia in Pediatric Patients in Serbia, Greece, and Romania. Current Status and Challenges for Collaboration

2020 ◽  
Vol 6 (2) ◽  
pp. 49
Author(s):  
Valentina Arsić Arsenijevic ◽  
Timoleon-Achilleas Vyzantiadis ◽  
Mihai Mares ◽  
Suzana Otasevic ◽  
Athanasios Tragiannidis ◽  
...  
Keyword(s):  
Molecular Diagnosis ◽  
Pediatric Patients ◽  
Fungal Infections ◽  
Laboratory Diagnosis ◽  
Pneumocystis Jirovecii ◽  
Current Status ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Clinical Practices ◽  
Online Questionnaire ◽  
Regional Collaboration

Pneumocystis jirovecii can cause fatal Pneumocystis pneumonia (PcP). Many children have been exposed to the fungus and are colonized in early age, while some individuals at high risk for fungal infections may develop PcP, a disease that is difficult to diagnose. Insufficient laboratory availability, lack of knowledge, and local epidemiology gaps make the problem more serious. Traditionally, the diagnosis is based on microscopic visualization of Pneumocystis in respiratory specimens. The molecular diagnosis is important but not widely used. The aim of this study was to collect initial indicative data from Serbia, Greece, and Romania concerning pediatric patients with suspected PcP in order to: find the key underlying diseases, determine current clinical and laboratory practices, and try to propose an integrative future molecular perspective based on regional collaboration. Data were collected by the search of literature and the use of an online questionnaire, filled by relevant scientists specialized in the field. All three countries presented similar clinical practices in terms of PcP prophylaxis and clinical suspicion. In Serbia and Greece the hematology/oncology diseases are the main risks, while in Romania HIV infection is an additional risk. Molecular diagnosis is available only in Greece. PcP seems to be under-diagnosed and regional collaboration in the field of laboratory diagnosis with an emphasis on molecular approaches may help to cover the gaps and improve the practices.

scholarly journals Evaluation of a Turbidimetric β-D-Glucan Test for Detection of Pneumocystis jirovecii Pneumonia

2018 ◽  
Vol 56 (7) ◽  
pp. e00286-18 ◽  
Author(s):  
Karl Dichtl ◽  
Ulrich Seybold ◽  
Johannes Wagener
Keyword(s):  
Sensitivity And Specificity ◽  
Test Performance ◽  
Laboratory Diagnosis ◽  
Turnover Time ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Content Type ◽  
Increased Risk ◽  
Turbidimetric Assay ◽  
Respiratory Specimens

ABSTRACT Currently, diagnosis of Pneumocystis jirovecii pneumonia (PJP) relies on analysis of lower respiratory specimens, either by microscopy or quantitative real-time PCR (qPCR). Thus, bronchoscopy is required, which is associated with increased risk of respiratory failure. We assessed the value of noninvasive serologic β-d-glucan (BDG) testing for laboratory diagnosis of PJP using a newly available turbidimetric assay. We identified 73 cases of PJP with positive qPCR results from lower respiratory specimens for Pneumocystis and serology samples dating from 1 week before to 4 weeks after qPCR. In addition, 25 sera from controls with suspected PJP but specimens negative for Pneumocystis by qPCR were identified. Sera were tested with a turbidimetric BDG assay (Fujifilm Wako Chemicals Europe GmbH, Neuss, Germany), using an 11-pg/ml cutoff. Sensitivity and specificity were calculated based on qPCR test results as a reference. The turbidimetric BDG assay identified 63/73 patients with positive or slightly positive qPCR tests for an overall sensitivity of 86%; after exclusion of cases with only slightly positive qPCR results, sensitivity was 91%. No correlation between serum BDG levels and respiratory specimen DNA levels was found. Serologic BDG testing was negative in 25/25 controls with negative qPCR for a specificity of 100% using the predefined cutoff. In 22/25 samples (88%), no BDG was detected. Serologic BDG testing using the turbidimetric assay showed high sensitivity and specificity compared to qPCR of lower respiratory specimens for the diagnosis of PJP. Both turnover time and test performance will allow clinicians to delay or in some cases forego bronchoscopy.

scholarly journals Performances of Four Real-Time PCR Assays for Diagnosis of Pneumocystis jirovecii Pneumonia

2015 ◽  
Vol 54 (3) ◽  
pp. 625-630 ◽  
Author(s):  
Milène Sasso ◽  
Elsa Chastang-Dumas ◽  
Sophie Bastide ◽  
Sandrine Alonso ◽  
Catherine Lechiche ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Essential Element ◽  
Laboratory Diagnosis ◽  
Final Diagnosis ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Content Type ◽  
Two Samples ◽  
Pcr Assays

Pneumonia due toPneumocystis jirovecii(PCP) is a frequent infection among HIV-positive or other immunocompromised patients. In the past several years, PCR on pulmonary samples has become an essential element for the laboratory diagnosis of PCP. Nevertheless, very few comparative studies of available PCR assays have been published. In this work, we evaluated the concordance between four real-time PCR assays, including three commercial kits, AmpliSens, MycAssay, and Bio-Evolution PCR, and an in-house PCR (J. Fillaux et al. 2008, J Microbiol Methods 75:258–261, doi:http://dx.doi.org/10.1016/j.mimet.2008.06.009), on 148 pulmonary samples. The results showed concordance rates ranging from 81.6% to 96.6% (kappa, 0.64 to 0.93). Concordance was excellent between three assays: the in-house assay, AmpliSens, and the MycAssay PCR (kappa, >0.8). The performances of these PCR assays were also evaluated according to the classification of the probability of PCP (proven, probable, possible, or no final diagnosis of PCP) based on clinical and radiological signs as well as on the direct examination of bronchoalveolar lavage samples. In the proven PCP category,Pneumocystis jiroveciiDNA was detected with all four assays. In the probable PCP category, the in-house PCR, AmpliSens, and the MycAssay PCR were positive for all samples, while the Bio-Evolution PCR failed to detectPneumocystis jiroveciiDNA in two samples. In the possible PCP category, the percentage of positive samples according to PCR varied from 54.5% to 86.4%. Detection of colonized patients is discussed. Finally, among the four evaluated PCR assays, one was not suitable for colonization detection but showed good performance in the proven and probable PCP groups. For the three other assays, performances were excellent and allowed detection of a very low fungal burden.

scholarly journals Pneumocystis jirovecii pneumonia: still a concern in patients with haematological malignancies and stem cell transplant recipients

2016 ◽  
Vol 71 (9) ◽  
pp. 2379-2385 ◽  
Author(s):  
Catherine Cordonnier ◽  
Simone Cesaro ◽  
Georg Maschmeyer ◽  
Hermann Einsele ◽  
J. Peter Donnelly ◽  
...  
Keyword(s):  
Stem Cell ◽  
Stem Cell Transplant ◽  
Laboratory Diagnosis ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Cell Transplant ◽  
Haematological Malignancies ◽  
Transplant Recipients ◽  
Allogeneic Hsct ◽  
Prevention And Treatment

The risk of patients with ALL and recipients of an allogeneic HSCT developing Pneumocystis jirovecii pneumonia is sufficiently high to warrant guidelines for the laboratory diagnosis, prevention and treatment of the disease. In this issue, the European Conference on Infections in Leukemia (ECIL) presents its recommendations in three companion papers.

scholarly journals Pneumocystis jirovecii Pneumonia: Current Advances in Laboratory Diagnosis

OBM Genetics ◽  
2018 ◽  
Vol 2 (4) ◽  
pp. 1-1 ◽  
Author(s):  
Ana Luísa Tomás ◽  
◽  
Olga Matos ◽  
◽  
◽  
...  
Keyword(s):  
Laboratory Diagnosis ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia

scholarly journals Molecular Diagnosis of Pneumocystis jirovecii Pneumonia by Use of Oral Wash Samples in Immunocompromised Patients: Usefulness and Importance of the DNA Target

2019 ◽  
Vol 57 (12) ◽  
Author(s):  
Lidia Goterris ◽  
Miguel Angel Mancebo Fernández ◽  
Juan Aguilar-Company ◽  
Vicenç Falcó ◽  
Isabel Ruiz-Camps ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Molecular Diagnosis ◽  
Small Subunit ◽  
Molecular Techniques ◽  
Pneumocystis Jirovecii ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Rrna Gene ◽  
Immunocompromised Patients ◽  
Content Type ◽  
Hiv Negative

ABSTRACT Pneumocystis jirovecii pneumonia (PJP) is an important cause of pneumonia in the HIV-negative immunocompromised population, for whom the fungal load is low, the differential diagnosis is difficult, and a bronchoalveolar lavage (BAL) sample is often not readily available. Molecular techniques have improved the microbiological diagnosis in this scenario. The usefulness of two real-time PCR techniques targeting nuclear single-copy and mitochondrial multicopy genes, respectively, applied to oral wash specimens (OW) for PJP diagnosis was assessed, and its accuracy was compared to a BAL fluid-based diagnosis. Immunocompromised patients having PJP in the differential diagnosis of an acute respiratory episode, and from whom OW and BAL or lung biopsy specimens were obtained ≤48 h apart, were retrospectively included. PCRs targeting the dihydropteroate synthase gene (DHPS) and the mitochondrial small-subunit (mtSSU) rRNA gene were performed in paired OW-BAL specimens. Thirty-six patients were included (88.6% HIV negative). Fifteen patients (41.7%) were classified as PJP, and a further 8 were considered P. jirovecii colonized. Quantification of DHPS and mtSSU in BAL fluid showed an accuracy of 96.9% and 93.0%, respectively, for PJP diagnosis, whereas a qualitative approach performed better when applied to OW (accuracy, 91.7%) irrespective of the PCR target studied (kappa = 1). Qualitative molecular diagnosis applied to OW showed an excellent performance for PJP diagnosis regardless of the target studied, being easier to interpret than the quantitative approach needed for BAL fluid.

scholarly journals Estimating the Burden of Serious Fungal Infections in Uruguay

2018 ◽  
Author(s):  
Marina Macedo-Viñas ◽  
David W. Denning
Keyword(s):  
Fungal Infections ◽  
Oral Candidiasis ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Laboratory Diagnosis ◽  
Population Characteristics ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Solid Organ ◽  
Hematopoietic Stem ◽  
Populations At Risk ◽  
Candida Peritonitis

We aimed to estimate for the first time the burden of fungal infections in Uruguay. Data on population characteristics and underlying conditions were extracted from the National Statistics Institute, the World Bank, national registries and published articles. When no data existed, risk populations were used to estimate frequencies extrapolating from the literature. Population structure: total 3,444,006; 73% adults; 35% women younger than 50 years. Size of populations at risk: HIV infected 12,000; acute myeloid leukemia 126; hematopoietic stem cell transplantation 30; solid organ transplants 134; COPD 272,006 (19.7% of older than 40); asthma in adults 223,431 (prevalence 9%); cystic fibrosis in adults 48; tuberculosis 613 (incidence 26.2%), lung cancer 1,400 (ASR incidence 27.4). Annual incidence estimations per 100,000: 22.4 invasive aspergillosis, 16.4 candidaemia, 3.7 Candida peritonitis, 1.62 Pneumocystis jirovecii pneumonia, 0.75 cryptococcosis, severe asthma with fungal sensitisation 217, allergic bronchopulmonary aspergillosis 165, recurrent Candida vaginitis 6,323, oral candidiasis 74.5 and oesophageal candidiasis 25.7. Although some under and overestimations could have been made, we expect that at least 127,525 people suffer from serious fungal infections each year. Sporothrichosis, histoplasmosis, paracoccidioidomycosis and dermatophytosis are known to be frequent but no data are available to make accurate estimations. Given the magnitude of the burden of fungal infections in Uruguay, efforts should be made to improve surveillance, strengthen laboratory diagnosis and warrant access to first line antifungals.

scholarly journals ECIL guidelines for the diagnosis of Pneumocystis jirovecii pneumonia in patients with haematological malignancies and stem cell transplant recipients

2016 ◽  
Vol 71 (9) ◽  
pp. 2386-2396 ◽  
Author(s):  
Alexandre Alanio ◽  
Philippe M. Hauser ◽  
Katrien Lagrou ◽  
Willem J. G. Melchers ◽  
Jannik Helweg-Larsen ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Pneumocystis Jirovecii ◽  
High Dose ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Cell Transplant ◽  
Haematological Malignancies ◽  
Early Management ◽  
Non Invasive ◽  
Patient At Risk

AbstractThe Fifth European Conference on Infections in Leukaemia (ECIL-5) convened a meeting to establish evidence-based recommendations for using tests to diagnose Pneumocystis jirovecii pneumonia (PCP) in adult patients with haematological malignancies. Immunofluorescence assays are recommended as the most sensitive microscopic method (recommendation A-II). Real-time PCR is recommended for the routine diagnosis of PCP (A-II). Bronchoalveolar lavage (BAL) fluid is recommended as the best specimen as it yields good negative predictive value (A-II). Non-invasive specimens can be suitable alternatives (B-II), acknowledging that PCP cannot be ruled out in case of a negative PCR result (A-II). Detecting β-d-glucan in serum can contribute to the diagnosis but not the follow-up of PCP (A-II). A negative serum β-d-glucan result can exclude PCP in a patient at risk (A-II), whereas a positive test result may indicate other fungal infections. Genotyping using multilocus sequence markers can be used to investigate suspected outbreaks (A-II). The routine detection of dihydropteroate synthase mutations in cases of treatment failure is not recommended (B-II) since these mutations do not affect response to high-dose co-trimoxazole. The clinical utility of these diagnostic tests for the early management of PCP should be further assessed in prospective, randomized interventional studies.

scholarly journals 837. Performance of Blood (1- >3)-β-D-Glucan in People with AIDS Presenting with Respiratory Symptoms

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S510-S511
Author(s):  
Lana Hasan ◽  
Gayathri Krishnan ◽  
Michael Saccente
Keyword(s):  
Bronchoalveolar Lavage ◽  
Diagnostic Test ◽  
Test Performance ◽  
Respiratory Symptoms ◽  
Fungal Infections ◽  
Pneumocystis Jirovecii ◽  
Cape Cod ◽  
Pneumocystis Jirovecii Pneumonia ◽  
Non Invasive ◽  
Progressive Disseminated Histoplasmosis

Abstract Background The gold standard for diagnosis of Pneumocystis jirovecii pneumonia (PCP) is direct visualization of the microorganism in respiratory samples, usually obtained via bronchoalveolar lavage (BAL). Blood β-D-glucan (BDG) is used as a non-invasive adjunctive diagnostic test for PCP, but specificity is only modest, in part because other opportunistic fungal infections cause high BDG. We previously showed BDG-positivity in 94% of people with AIDS (PWA), progressive disseminated histoplasmosis (PDH), and respiratory symptoms in our hospital. In this study, we aim to assess the performance of BDG as a diagnostic test for PCP in PWA who have respiratory symptoms. Methods We retrospectively identified PWA who had a BDG result between 2014 and 2019. AIDS was defined as past or current absolute CD4 count < 200 cells/µL, or a past or current AIDS-defining condition. Positive cytological or histological evidence of P. jirovecii in bronchoalveolar lavage (BAL) fluid or lung biopsy, or positive Pneumocystis PCR on sputum or BAL confirmed PCP. The Fungitell Assay (Associates of Cape Cod, East Falmouth, MA) determined BDG levels as follows: negative, < 60 pg/mL; indeterminate, 60-79 pg/mL, and positive, ≥ 80 pg/mL. Values < 31 pg/mL and those >500 pg/mL were censored at 30 pg/mL and 500 pg/mL, respectively. Respiratory symptoms were defined as cough, dyspnea, chest pain, or hypoxia. We compared BDG results for participants with proven PCP and participants without proven PCP. Results We identified 260 PWA with a BDG result, of whom 183 had at least one respiratory symptom. 84 (45.9%) of these participants had a positive BDG. BDG results among participants with and without PCP are shown in Table 1. Of the 44 participants with a positive BDG who did not have PCP, 29 (65.9%) had PDH. Other diagnoses included cryptococcosis and candidemia. The test performance of BDG for the diagnosis of PCP is shown in Table 2. Exclusion of participants with PDH increased the specificity of BDG for PCP to 86.4%. Table 1. Results of (1->3)-β-D-glucan Testing by Pneumocystis jirovecii Pneumonia Diagnosis Among Participants with AIDS and Respiratory Symptoms Table 2. Test Performance of (1->3)-β-D-glucan for the Diagnosis of Pneumocystis jirovecii Pneumonia* Conclusion At our center where histoplasmosis is endemic, a positive BDG should not be attributed to PCP among PWA with respiratory symptoms because of low specificity and low positive predictive value. However, a negative BDG can exclude PCP in this population. Disclosures All Authors: No reported disclosures

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