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
• bronchoalveolární lavážní tekutina mikrobiologie   MeSH
• imunokompromitovaný pacient MeSH
• imunosupresivní léčba škodlivé účinky   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé MeSH
• Pneumocystis carinii * izolace a purifikace   MeSH
• pneumocystová pneumonie diagnóza   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Slovenská republika MeSH

• MeSH
• imunosupresivní léčba škodlivé účinky   MeSH
• lidé MeSH
• Pneumocystis carinii * patogenita   MeSH
• pneumocystová pneumonie diagnóza   farmakoterapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• akutní retinální nekróza MeSH
• brucelóza farmakoterapie   krev   patofyziologie   MeSH
• chorioretinitida MeSH
• diferenciální diagnóza MeSH
• herpetické infekce farmakoterapie   klasifikace   krev   patofyziologie   MeSH
• histoplazmóza farmakoterapie   imunologie   patofyziologie   MeSH
• infekce bakteriemi rodu Bartonella farmakoterapie   krev   patofyziologie   MeSH
• infekční nemoci * klasifikace   komplikace   MeSH
• kandidóza farmakoterapie   patofyziologie   patologie   MeSH
• kryptokokóza farmakoterapie   imunologie   komplikace   MeSH
• leptospiróza diagnóza   farmakoterapie   mikrobiologie   patofyziologie   MeSH
• lidé MeSH
• lymeská nemoc farmakoterapie   krev   patofyziologie   MeSH
• myiáza diagnostické zobrazování   klasifikace   přenos   MeSH
• oční symptomy * MeSH
• oční toxoplazmóza diagnostické zobrazování   farmakoterapie   MeSH
• onchocerkóza oční diagnostické zobrazování   MeSH
• Pneumocystis carinii izolace a purifikace   patogenita   účinky léků   MeSH
• schistosomóza diagnostické zobrazování   klasifikace   přenos   MeSH
• spalničky komplikace   krev   patofyziologie   MeSH
• toxokaróza imunologie   patologie   MeSH
• toxoplazmóza MeSH
• uveitida klasifikace   MeSH
• zánět zrakového nervu MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy 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
• dospělí MeSH
• HIV infekce * komplikace   farmakoterapie   MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• Pneumocystis carinii * MeSH
• pneumocystová pneumonie * epidemiologie   prevence a kontrola   MeSH
• počet CD4 lymfocytů MeSH
• viremie epidemiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

• MeSH
• antiretrovirové látky terapeutické užití   MeSH
• dospělí MeSH
• HIV infekce * diagnóza   MeSH
• klinické laboratorní techniky metody   MeSH
• lidé MeSH
• mozková toxoplazmóza * diagnóza   farmakoterapie   MeSH
• neurozobrazování metody   MeSH
• Pneumocystis carinii izolace a purifikace   MeSH
• pneumocystová pneumonie diagnóza   farmakoterapie   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• kazuistiky 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
• bronchoalveolární lavážní tekutina mikrobiologie   MeSH
• diagnostické techniky molekulární metody   normy   MeSH
• DNA fungální genetika   MeSH
• kvantitativní polymerázová řetězová reakce normy   MeSH
• lidé MeSH
• Pneumocystis carinii genetika   MeSH
• pneumocystová pneumonie diagnóza   mikrobiologie   MeSH
• senzitivita a specificita MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• multicentrická studie 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 genetika   izolace a purifikace   patogenita   MeSH
• bronchoalveolární lavážní tekutina mikrobiologie   MeSH
• DNA fungální genetika   MeSH
• dospělí MeSH
• hematologické nádory komplikace   mikrobiologie   MeSH
• jednotky intenzivní péče MeSH
• lidé středního věku MeSH
• lidé MeSH
• mannany analýza   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• neutropenie mikrobiologie   MeSH
• Pneumocystis carinii genetika   izolace a purifikace   patogenita   MeSH
• pneumocystová pneumonie diagnóza   mikrobiologie   MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Streptococcus pneumoniae genetika   izolace a purifikace   patogenita   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články 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
• biologická evoluce MeSH
• fylogeneze MeSH
• geny hub MeSH
• interakce hostitele a patogenu MeSH
• Muridae mikrobiologie   MeSH
• plíce mikrobiologie   MeSH
• Pneumocystis klasifikace   genetika   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Afrika MeSH

• MeSH
• Aspergillus fumigatus patogenita   MeSH
• bakteriální infekce diagnóza   terapie   MeSH
• Candida albicans patogenita   MeSH
• Cryptococcus gattii patogenita   MeSH
• Cryptococcus neoformans patogenita   MeSH
• cytomegalovirové infekce diagnostické zobrazování   diagnóza   terapie   MeSH
• herpetické infekce diagnóza   farmakoterapie   MeSH
• infekce virem Epsteina-Barrové diagnóza   MeSH
• infekce diagnóza   terapie   MeSH
• lidé MeSH
• Mucorales patogenita   MeSH
• mykózy diagnostické zobrazování   diagnóza   terapie   MeSH
• Pneumocystis patogenita   MeSH
• Strongyloides patogenita   MeSH
• Toxoplasma patogenita   MeSH
• transplantace plic * MeSH
• Check Tag
• lidé 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
• fylogeneze MeSH
• genetické markery MeSH
• genotyp MeSH
• hlístice genetika   MeSH
• hybridizace genetická * MeSH
• mitochondriální DNA genetika   MeSH
• myši genetika   parazitologie   MeSH
• paraziti genetika   MeSH
• Pneumocystis genetika   MeSH
• populační genetika * MeSH
• zvířata MeSH
• Check Tag
• myši genetika   parazitologie   MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Německo MeSH