Cieľ: Pneumocystis jirovecii patril v minulosti do skupiny prvokov, ale v súčasnosti je taxonomicky zaradený do ríše húb. P. jirovecii je oportúnny patogén, zodpovedný za pneumocystovú pneumóniu s častými komplikáciami u imunokompromitovaných pacientov. Oneskorené začatie vhodnej liečby zvyšuje riziko úmrtia u pacientov s oslabenou imunitou. Cieľom práce bolo zistiť a zhodnotiť spoľahlivosť metód laboratórnej diagnostiky pneumocystózy používaných v rutinných laboratóriách ako aj výskyt tohto ochorenia u pacientov zo Slovenska za 19 rokov. Materiál a metódy: Diagnostika je založená na mikroskopickom dôkaze (farbenie podľa Giemsa a Gram-Weigerta) a detekcii DNA parazita klasickou alebo real-time PCR v bronchoalveolárnej laváži a spúte. Výsledky: Pneumocysty boli zistené u 190 osôb (5,7 %) z celého súboru pacientov. Onkologickí pacienti predstavovali najrizikovejšiu skupinu z hľadiska infekcie pneumocystami, čo sme potvrdili ich najvyšším podielom (57,9 %) z jedincov s pneumocystózou. Na základe binárneho klasifikačného testu sme vyhodnotili 33,7 % citlivosť a 100 % špecifickosť mikroskopického dôkazu v porovnaní s PCR. Molekulárne metódy sú v porovnaní s mikroskopickým dôkazom citlivejšie v detekcii P. jirovecii a v súčasnosti predstavujú spoľahlivý detekčný systém v diagnostike pneumocystózy. Záver: Vzhľadom na narastajúci počet imunokompromitovaných osôb je diagnostika P. jirovecii u pacientov s pľúcnymi komplikáciami nevyhnutná. To sa potvrdilo aj v našej štúdii, kde v priebehu rokov stúpal počet vyšetrení a záchytov tohto oportúnneho patogénu.

Aim: In the past, Pneumocystis jirovecii belonged to the Protozoa group, but is currently taxonomically included in the kingdom Fungi. P. jirovecii is an opportunistic pathogen, responsible for pneumocystis pneumonia with frequent complications of immunocompromised patients. Delayed initiation of appropriate therapy increases the risk of death in immunocompromised patient. The aim of this work was to determine and evaluate the reliability of methods of laboratory diagnosis of pneumocystosis used in routine laboratories as well as the occurrence of this disease in patients from Slovakia during 19 years. Material and Methods: The diagnosis is based on microscopic examination (Giemsa- and Gram-Weigert-staining) and detection of parasite DNA by classical or real-time PCR in bronchoalveolar lavage and sputum. Results: Pneumocysts were detected in 190 persons (5.7%) from the whole group of patients. Cancer patients represented the riskiest group in terms of pneumocystosis, which was confirmed by the highest percentage (57.9%) of individuals infected with P. jirovecii. Compared with the PCR, 33.7% sensitivity and 100% specificity of microscopy was calculated by using a binary classification test. Molecular methods are more sensitive in the detection of P. jirovecii compared to microscopic evidence and currently represent a reliable detection system in the diagnosis of pneumocystosis. Conclusion: In view of the increasing number of immunocompromised persons, diagnostics of P. jirovecii in patients with pulmonary complications is essential. This was also confirmed in our study, where the number of examinations and detection of this opportunistic pathogen increased over the years.

• MeSH
• Bronchoalveolar Lavage Fluid microbiology   MeSH
• Immunocompromised Host MeSH
• Immunosuppression Therapy adverse effects   MeSH
• Real-Time Polymerase Chain Reaction MeSH
• Humans MeSH
• Pneumocystis carinii * isolation & purification   MeSH
• Pneumonia, Pneumocystis diagnosis   microbiology   MeSH
• Check Tag
• Humans MeSH
• Geographicals
• Slovakia MeSH

• MeSH
• Immunosuppression Therapy adverse effects   MeSH
• Humans MeSH
• Pneumocystis carinii * pathogenicity   MeSH
• Pneumonia, Pneumocystis diagnosis   drug therapy   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Retinal Necrosis Syndrome, Acute MeSH
• Brucellosis drug therapy   blood   physiopathology   MeSH
• Chorioretinitis MeSH
• Diagnosis, Differential MeSH
• Herpesviridae Infections drug therapy   classification   blood   physiopathology   MeSH
• Histoplasmosis drug therapy   immunology   physiopathology   MeSH
• Bartonella Infections drug therapy   blood   physiopathology   MeSH
• Communicable Diseases * classification   complications   MeSH
• Candidiasis drug therapy   physiopathology   pathology   MeSH
• Cryptococcosis drug therapy   immunology   complications   MeSH
• Leptospirosis diagnosis   drug therapy   microbiology   physiopathology   MeSH
• Humans MeSH
• Lyme Disease drug therapy   blood   physiopathology   MeSH
• Myiasis diagnostic imaging   classification   transmission   MeSH
• Eye Manifestations * MeSH
• Toxoplasmosis, Ocular diagnostic imaging   drug therapy   MeSH
• Onchocerciasis, Ocular diagnostic imaging   MeSH
• Pneumocystis carinii isolation & purification   pathogenicity   drug effects   MeSH
• Schistosomiasis diagnostic imaging   classification   transmission   MeSH
• Measles complications   blood   physiopathology   MeSH
• Toxocariasis immunology   pathology   MeSH
• Toxoplasmosis MeSH
• Uveitis classification   MeSH
• Optic Neuritis MeSH
• Check Tag
• Humans MeSH
• Publication type
• Review MeSH

INTRODUCTION: Since the beginning of the HIV epidemic in resource-rich countries, Pneumocystis jirovecii pneumonia (PjP) is one of the most frequent opportunistic AIDS-defining infections. The Collaboration of Observational HIV Epidemiological Research Europe (COHERE) has shown that primary Pneumocystis jirovecii Pneumonia (PjP) prophylaxis can be safely withdrawn in patients with CD4 counts of 100 to 200 cells/µL if plasma HIV-RNA is suppressed on combination antiretroviral therapy. Whether this holds true for secondary prophylaxis is not known, and this has proved difficult to determine due to the much lower population at risk. METHODS: We estimated the incidence of secondary PjP by including patient data collected from 1998 to 2015 from the COHERE cohort collaboration according to time-updated CD4 counts, HIV-RNA and use of PjP prophylaxis in persons >16 years of age. We fitted a Poisson generalized additive model in which the smoothed effect of CD4 was modelled by a restricted cubic spline, and HIV-RNA was stratified as low (<400), medium (400 to 10,000) or high (>10,000copies/mL). RESULTS: There were 373 recurrences of PjP during 74,295 person-years (py) in 10,476 patients. The PjP incidence in the different plasma HIV-RNA strata differed significantly and was lowest in the low stratum. For patients off prophylaxis with CD4 counts between 100 and 200 cells/µL and HIV-RNA below 400 copies/mL, the incidence of recurrent PjP was 3.9 (95% CI: 2.0 to 5.8) per 1000 py, not significantly different from patients on prophylaxis in the same stratum (1.9, 95% CI: 0.1 to 3.7). CONCLUSIONS: HIV viraemia importantly affects the risk of recurrent PjP. In virologically suppressed patients on ART with CD4 counts of 100 to 200/µL, the incidence of PjP off prophylaxis is below 10/1000 py. Secondary PjP prophylaxis may be safely withheld in such patients. While European guidelines recommend discontinuing secondary PjP prophylaxis only if CD4 counts rise above 200 cells/mL, the latest US Guidelines consider secondary prophylaxis discontinuation even in patients with a CD4 count above 100 cells/µL and suppressed viral load. Our results strengthen and support this US recommendation.

• MeSH
• Adult MeSH
• HIV Infections * complications   drug therapy   MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Pneumocystis carinii * MeSH
• Pneumonia, Pneumocystis * epidemiology   prevention & control   MeSH
• CD4 Lymphocyte Count MeSH
• Viremia epidemiology   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Publication type
• Journal Article MeSH
• Observational Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Europe MeSH

• MeSH
• Anti-Retroviral Agents therapeutic use   MeSH
• Adult MeSH
• HIV Infections * diagnosis   MeSH
• Clinical Laboratory Techniques methods   MeSH
• Humans MeSH
• Toxoplasmosis, Cerebral * diagnosis   drug therapy   MeSH
• Neuroimaging methods   MeSH
• Pneumocystis carinii isolation & purification   MeSH
• Pneumonia, Pneumocystis diagnosis   drug therapy   MeSH
• Treatment Outcome MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Case Reports MeSH

Quantitative real-time PCR (qPCR) is increasingly used to detect Pneumocystis jirovecii for the diagnosis of Pneumocystis pneumonia (PCP), but there are differences in the nucleic acids targeted, DNA only versus whole nucleic acid (WNA), and also the target genes for amplification. Through the Fungal PCR Initiative, a working group of the International Society for Human and Animal Mycology, a multicenter and monocenter evaluation of PCP qPCR assays was performed. For the multicenter study, 16 reference laboratories from eight different countries, performing 20 assays analyzed a panel consisting of two negative and three PCP positive samples. Aliquots were prepared by pooling residual material from 20 negative or positive- P. jirovecii bronchoalveolar lavage fluids (BALFs). The positive pool was diluted to obtain three concentrations (pure 1:1; 1:100; and 1:1000 to mimic high, medium, and low fungal loads, respectively). The monocenter study compared five in-house and five commercial qPCR assays testing 19 individual BALFs on the same amplification platform. Across both evaluations and for all fungal loads, targeting WNA and the mitochondrial small sub-unit (mtSSU) provided the earliest Cq values, compared to only targeting DNA and the mitochondrial large subunit, the major surface glycoprotein or the beta-tubulin genes. Thus, reverse transcriptase-qPCR targeting the mtSSU gene could serve as a basis for standardizing the P. jirovecii load, which is essential if qPCR is to be incorporated into clinical care pathways as the reference method, accepting that additional parameters such as amplification platforms still need evaluation.

• MeSH
• Bronchoalveolar Lavage Fluid microbiology   MeSH
• Molecular Diagnostic Techniques methods   standards   MeSH
• DNA, Fungal genetics   MeSH
• Real-Time Polymerase Chain Reaction standards   MeSH
• Humans MeSH
• Pneumocystis carinii genetics   MeSH
• Pneumonia, Pneumocystis diagnosis   microbiology   MeSH
• Sensitivity and Specificity MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Multicenter Study MeSH

The present study aims to evaluate the diagnostic yield of bronchoalveolar lavage (BAL) fluid in patients with hematological malignancies and describe the most common pathogens detected in BAL fluid (BALF.) An analysis of 480 BALF samples was performed in patients with hematological malignancies over a period of 7 years. The results of culture methods, PCR, and immunoenzymatic sandwich microplate assays for Aspergillus galactomannan (GM) in BALF were analyzed. Further, the diagnostic thresholds for Aspergillus GM and Pneumocystis jiroveci were also calculated. Microbiological findings were present in 87% of BALF samples. Possible infectious pathogens were detected in 55% of cases; 32% were classified as colonizing. No significant difference in diagnostic yield or pathogen spectrum was found between non-neutropenic and neutropenic patients. There was one significant difference in BALF findings among intensive care units (ICU) versus non-ICU patients for Aspergillus spp. (22% versus 9%, p = 0.03). The most common pathogens were Aspergillus spp. (n = 86, 33% of BAL with causative pathogens) and Streptococcus pneumoniae (n = 46, 18%); polymicrobial etiology was documented in 20% of cases. A quantitative PCR value of > 1860 cp/mL for Pneumocystis jirovecii was set as a diagnostic threshold for pneumocystis pneumonia. The absorbance index of GM in BALF of 0.5 was set as a diagnostic threshold for aspergillosis. The examination of BAL fluid revealed the presence of pathogen in more than 50% of cases and is, therefore, highly useful in this regard when concerning pulmonary infiltrates.

• MeSH
• Aspergillus genetics   isolation & purification   pathogenicity   MeSH
• Bronchoalveolar Lavage Fluid microbiology   MeSH
• DNA, Fungal genetics   MeSH
• Adult MeSH
• Hematologic Neoplasms complications   microbiology   MeSH
• Intensive Care Units MeSH
• Middle Aged MeSH
• Humans MeSH
• Mannans analysis   MeSH
• Adolescent MeSH
• Young Adult MeSH
• Neutropenia microbiology   MeSH
• Pneumocystis carinii genetics   isolation & purification   pathogenicity   MeSH
• Pneumonia, Pneumocystis diagnosis   microbiology   MeSH
• Retrospective Studies MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Streptococcus pneumoniae genetics   isolation & purification   pathogenicity   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Male MeSH
• Aged, 80 and over MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Pneumocystis is a genus of parasitic fungi infecting lung tissues in a wide range of mammal species, displaying a strong host specificity and patterns of co-speciation with their hosts. However, a recent study on Asiatic murids challenged these patterns reporting several Pneumocystis lineages/species shared by different host species or even genera in the Rattini and Murini tribes. Here we screened lung samples of 27 species of African rodents from five families for the presence of Pneumocystis DNA. Using reconstructed multi-locus phylogenies of both hosts and parasites, we tested the hypothesis of their co-evolution. We found that Pneumocystis is widespread in African rodents, detected in all but seven screened host species, with species-level prevalence ranging from 5.9 to 100%. Several host species carry pairs of highly divergent Pneumocystis lineages/species. The retrieved co-phylogenetic signal was highly significant (p = .0017). We found multiple co-speciations, sorting events and two host-shift events, which occurred between Murinae and Deomyinae hosts. Comparison of genetic distances suggests higher substitution rates for Pneumocystis relative to the rodent hosts on neutral loci and slower rates on selected ones. We discuss life-history traits and population dynamics factors which could explain the observed results.

• MeSH
• Biological Evolution MeSH
• Phylogeny MeSH
• Genes, Fungal MeSH
• Host-Pathogen Interactions MeSH
• Muridae microbiology   MeSH
• Lung microbiology   MeSH
• Pneumocystis classification   genetics   physiology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Africa MeSH

• MeSH
• Aspergillus fumigatus pathogenicity   MeSH
• Bacterial Infections diagnosis   therapy   MeSH
• Candida albicans pathogenicity   MeSH
• Cryptococcus gattii pathogenicity   MeSH
• Cryptococcus neoformans pathogenicity   MeSH
• Cytomegalovirus Infections diagnostic imaging   diagnosis   therapy   MeSH
• Herpesviridae Infections diagnosis   drug therapy   MeSH
• Epstein-Barr Virus Infections diagnosis   MeSH
• Infections diagnosis   therapy   MeSH
• Humans MeSH
• Mucorales pathogenicity   MeSH
• Mycoses diagnostic imaging   diagnosis   therapy   MeSH
• Pneumocystis pathogenicity   MeSH
• Strongyloides pathogenicity   MeSH
• Toxoplasma pathogenicity   MeSH
• Lung Transplantation * MeSH
• Check Tag
• Humans MeSH

Parasite hybrid zones resulting from host secondary contact have never been described in nature although parasite hybridization is well known and secondary contact should affect them similarly to free-living organisms. When host populations are isolated, diverge and recontact, intimate parasites (host specific, direct life cycle) carried during isolation will also meet and so may form parasite hybrid zones. If so, we hypothesize these should be narrower than the host's hybrid zone as shorter parasite generation time allows potentially higher divergence. We investigate multilocus genetics of two parasites across the European house mouse hybrid zone. We find each host taxon harbours its own parasite taxa. These also hybridize: Parasite hybrid zones are significantly narrower than the host's. Here, we show a host hybrid zone is a suture zone for a subset of its parasite community and highlight the potential of such systems as windows on the evolutionary processes of host-parasite interactions and recombinant pathogen emergence.

• MeSH
• Phylogeny MeSH
• Genetic Markers MeSH
• Genotype MeSH
• Nematoda genetics   MeSH
• Hybridization, Genetic * MeSH
• DNA, Mitochondrial genetics   MeSH
• Mice genetics   parasitology   MeSH
• Parasites genetics   MeSH
• Pneumocystis genetics   MeSH
• Genetics, Population * MeSH
• Animals MeSH
• Check Tag
• Mice genetics   parasitology   MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH
• Germany MeSH