Invasive pulmonary aspergillosis (IPA) may be a rare cause of granulomatous pneumonia in horses. The mortality of IPA is almost 100%; direct diagnostic tools in horses are needed. Bronchoalveolar lavage fluid (BALF) and serum samples were collected from 18 horses, including individuals suffering from IPA (n = 1), equine asthma (EA, n = 12), and 5 healthy controls. Serum samples were collected from another 6 healthy controls. Samples of BALF (n = 18) were analyzed for Aspergillus spp. DNA, fungal galactomannan (GM), ferricrocin (Fc), triacetylfusarinin C (TafC), and gliotoxin (Gtx). Analysis of 24 serum samples for (1,3)-β-D-glucan (BDG) and GM was performed. Median serum BDG levels were 131 pg/mL in controls and 1142 pg/mL in IPA. Similar trends were observed in BALF samples for GM (Area under the Curve (AUC) = 0.941) and DNA (AUC = 0.941). The fungal secondary metabolite Gtx was detected in IPA BALF and lung tissue samples (86 ng/mL and 2.17 ng/mg, AUC = 1).

• Publikační typ
• časopisecké články MeSH

The multiple forms of pulmonary aspergillosis caused by Aspergillus species are the most common respiratory mycoses. Although invasive, the analysis of diagnostic biomarkers in bronchoalveolar lavage fluid (BALF) is a clinical standard for diagnosing these conditions. The BALF samples from 22 patients with proven or probable aspergillosis were assayed for human pentraxin 3 (Ptx3), fungal ferricrocin (Fc), and triacetylfusarinine C (TafC) in a retrospective study. The infected group included patients with invasive pulmonary aspergillosis (IPA) and chronic aspergillosis (CPA). The BALF data were compared to a control cohort of 67 patients with invasive pulmonary mucormycosis (IPM), non-Aspergillus colonization, or bacterial infections. The median Ptx3 concentrations in patients with and without aspergillosis were 4545.5 and 242.0 pg/mL, respectively (95% CI, p < 0.05). The optimum Ptx3 cutoff for IPA was 2545 pg/mL, giving a sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 100, 98, 95, and 100%, respectively. The median Ptx3 concentration for IPM was high at 4326 pg/mL. Pentraxin 3 assay alone can distinguish IPA from CPA and invasive fungal disease from colonization. Combining Ptx3 and TafC assays enabled the diagnostic discrimination of IPM and IPA, giving a specificity and PPV of 100%.

• Publikační typ
• časopisecké články MeSH

Onychomycosis is one of the most common nail disorders. Its current treatment is not satisfactorily effective and often causes adverse side effects. This study aims to determine the optimal conditions for non-thermal plasma (NTP) inactivation of the most common dermatophytes in vitro and to apply it in patient`s therapy. The in vitro exposure to NTP produced by negative DC corona discharge caused full inactivation of Trichophyton spp. if applied during the early growth phases. This effect decreased to negligible inactivation with the exposure applied six days after inoculation. In a group of 40 patients with onychomycosis, NTP therapy was combined with nail plate abrasion and refreshment (NPAR) or treatment with antimycotics. The cohort included 17 patients treated with NPAR combined with NTP, 11 patients treated with antimycotics and NTP, and 12 patients treated with NPAR alone. The combination of NPAR and NTP resulted in clinical cure in more than 70% of patients. The synergistic effect of NPAR and NTP caused 85.7% improvement of mycological cure confirmed by negative microscopy and culture of the affected nail plate. We conclude that NTP can significantly improve the treatment of onychomycosis.

• MeSH
• Arthrodermataceae izolace a purifikace   MeSH
• lidé MeSH
• mykologické určovací techniky metody   MeSH
• onychomykóza * terapie   MeSH
• plazmové plyny terapeutické užití   MeSH
• techniky in vitro MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

Kryptické druhy sekce Fumigati, tzn. druhy podobné Aspergillus fumigatus, jsou stále častěji uváděny v literatuře jako původci invazivní aspergilózy (IA) u lidí i zvířat. Jejich odhalení a správná identifikace je důležitá, avšak ještě důležitější je stanovení citlivosti daného izolátu k antimykotikům (minimální inhibiční koncentrace, MIC) za použití vhodných metod. Pro tyto kryptické druhy jsou totiž často charakteristické zvýšené hodnoty MIC k lékům volby doporučeným pro terapii IA, jako je vorikonazol nebo amfotericin B.V našem sdělení uvádíme případ plicní aspergilózy u 63letého muže po transplantaci srdce. Jako původce infekce byl kultivačně prokázán a pomocí sekvenace DNA identifikován A. lentulus, se sníženou citlivostí k vorikonazolu a amfotericinu B. Citlivost k antifungálním látkám byla ověřena pomocí standardizované metodiky EUCAST-AFST. Terapie byla na základě získaných hodnot MIC cíleně změněna z vorikonazolu na posakonazol s výborným klinickým efektem. Pokud víme, naše kazuistika je prvním popsaným případem terapie A. lentulusposakonazolem a navíc úspěšným.

Cryptic species within the section Fumigati, that is Aspergillus fumigatus-like species, are increasingly reported in the literature as causative agents of invasive aspergillosis (IA) in both humans and animals. Their detection and proper identification are important, buteven more important is to determine the susceptibility profile (minimum inhibitory concentrations, MICs) of the isolate to antifungalsusing appropriate methods. Cryptic species often demonstrate elevated MICs to drugs recommended for IA therapy such as voriconazole or amphotericin B. Presented is a case of pulmonary aspergillosis in a 63-year-old male heart transplant recipient. Aspergilluslentuluswith reduced susceptibility to voriconazole and amphotericin B was identified as the causative agent of the infection using culture and DNA sequencing. Susceptibility to antifungals was confirmed by the standard EUCAST-AFST methods. Based on MIC values obtained in vitro, therapy was switched from voriconazole to posaconazole with excellent clinical effects. To the best of our knowledge, this is the first reported case of A. lentulusinfection treated with posaconazole and, moreover, a successful one.

Cerebral abscesses caused by dark-pigmented Fonsecaea fungi are rare, especially in otherwise healthy individuals. In this case report, we present a 61-year-old man from Moldova, living in the Czech Republic, who had worked as a locksmith on oil platforms in Turkmenistan, Kazakhstan, Sudan, and Iraq since 1999, and was admitted to a neurology ward for a sudden motion disorder of the right leg, dysarthria, and hypomimia. Imaging revealed presence of expansive focus around the left lateral ventricle of the brain and a pronounced peripheral edema. The intracranial infectious focus was excised under intraoperative SonoWand guidance. Tissue samples were histologically positive for dark-pigmented hyphae, suggesting dematiaceous fungi. Therefore, liposomal amphotericin B therapy was initiated immediately. Fonsecaea monophora was provisionally identified using ITS rDNA region sequencing directly from brain tissue. The identification was subsequently confirmed by cultivation and DNA sequencing from culture. The strain exhibited in vitro sensitive to voriconazole (MIC = 0.016 μg/mL) and resistance to amphotericin B (MIC = 4 μg/mL); therefore, the amphotericin B was replaced with voriconazole. Postoperatively, a significant clinical improvement was observed and no additional surgery was required. Based on the literature review, this is the third documented case of cerebral infection due to this pathogen in patients without underlying conditions and the first such case in Europe.

• MeSH
• absces mozku diagnostické zobrazování   mikrobiologie   chirurgie   MeSH
• amfotericin B terapeutické užití   MeSH
• antifungální látky farmakologie   terapeutické užití   MeSH
• Ascomycota účinky léků   genetika   izolace a purifikace   MeSH
• imunokompetence MeSH
• lidé středního věku MeSH
• lidé MeSH
• mykózy diagnóza   diagnostické zobrazování   MeSH
• ribozomální DNA genetika   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• přehledy MeSH
• Geografické názvy
• Česká republika MeSH

Detection of serum galactomannan (GM) and (1,3)-β-d-glucan (BG) is considered useful for non-culture diagnosis of invasive pulmonary aspergillosis (IPA) in neutropenic patients. Only few studies evaluated these seromarkers in non-neutropenic patients suspected of having IPA. The aim of this study was to evaluate both tests together with the Aspergillus fumigatus-specific serum IgG and IgA (IgAG) test for serological IPA diagnosis in non-neutropenic patients. Sera from 87 patients suspected of having IPA were retrospectively analysed. Patients were categorised into groups of proven IPA (n = 10), putative IPA (n = 31) and non-IPA colonisation (n = 46). When the GM, BG and IgAG assays were used for patients included in the study, the sensitivity/specificity/positive predictive value (PPV)/negative predictive value (NPV) were 48.8%/91.3%/83.3%/66.7%, 82.9%/73.9%/73.9%/82.9% and 75.6%/95.7%/93.9%/81.5%, respectively. Thus, the highest specificity and PPV were confirmed for the IgAG assay. Improvements in the sensitivity and NPV were achieved by "at least one positive" analysis with the GM and BG assays, with the sensitivity/specificity/PPV/NPV values being 85.0%/69.6%/71.4%/84.2%. Nevertheless, the highest sensitivity and NPV were achieved by the "at least one positive" analysis combining the GM, BG and IgAG tests (97.6% and 96.8%, respectively). The involvement of the IgAG assay could improve IPA diagnosis in non-neutropenic patients by increasing the sensitivity and NPV when combined with the GM or BG assays. Furthermore, improvement was achieved by combining the GM, BG and IgAG assays using the "at least one positive test" strategy, especially if doubt exists.

• MeSH
• Aspergillus fumigatus chemie   imunologie   MeSH
• beta-glukany krev   MeSH
• dospělí MeSH
• imunoglobulin A krev   MeSH
• imunoglobulin G krev   MeSH
• invazivní plicní aspergilóza diagnóza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mannany krev   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• prediktivní hodnota testů MeSH
• protilátky fungální krev   MeSH
• retrospektivní studie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• sérum chemie   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
• hodnotící studie MeSH
• multicentrická studie MeSH

Cílem studie bylo otestovat použitelnost metody PCR v kombinaci s High Resolution Melting Analysis (HRMA) pro detekci a identifikaci dermatofytů přímo z klinických vzorků kůže a jejích adnex. Metodika by měla vést ke zrychlení a zvýšení výtěžnosti diagnostiky dermatomykóz a ke zkvalitnění péče o pacienta. Ve studii bylo analyzováno 128 klinických vzorků od pacientů s podezřením na dermatomykózu. Pro izolaci DNA dermatofytů z klinického materiálu byl použit izolační KIT ZR Fungal/Bacterial DNA MiniPrepTM. Pro detekci DNA oblastí kódujících ribozomální RNA byly využity dvě sady primerů zachycující široké spektrum dermatofytů. Pro druhovou identifikaci dermatofytů byla použita real-time PCR metoda v kombinaci s High Resolution Melting Analysis (PCR-HRMA). Dermatofytóza byla potvrzena u 42 ze 128 zkoumaných vzorků (pozitivní přímá mikroskopie, kultivace, detekce PCR-HRMA, nebo kombinace metod). Výtěžnost PCR detekce byla 74 % pro obě sady primerů, metoda HRMA umožnila ve všech případech druhovou identifikaci detekovaných dermatofytů pomocí PCR. Naproti tomu kultivace byla pozitivní pouze u 52 % pacientů s dermatofytózou. Mikroskopie ze vzorku byla pozitivní v 90 % pozitivních dermatofytóz. Při současném použití mikroskopie, kultivace a PCR-HRMA bylo úspěšně identifikováno na úroveň druhu 90 % dermatofytů. Nejčastější druhy dermatofytů (Trichophyton rubrum, T. interdigitale, T. benhamiae) byly touto metodikou spolehlivě zachyceny. PCR detekce rDNA přímo z klinického materiálu s využitím HRMA zvýšila výtěžnost diagnostiky dermatofytóz zvláště tím, že přispěla k dosažení druhové identifikace u výrazně vyššího počtu případů v porovnání s konvenčními metodami. Kombinace konvenčních a molekulárně biologických vyšetření se zdá být vhodná pro rychlou a spolehlivou diagnostiku dermatofytóz bez nutnosti výrazného navýšení nákladů.

The aim of the study was to test the utility of the PCR method in combination with High Resolution Melting Analysis (HRMA) for the detection and identification of dermatophytes directly from clinical skin and adnexa samples. The methodology should reduce the time between sampling and diagnosis, increase the diagnostic sensitivity, and overall improve patient care. In the study, 128 clinical samples from patients with suspected dermatomycosis were analyzed. To isolate fungal DNA from the clinical specimens, a KIT ZR Fungal/Bacterial DNA MiniPrepTM was used. Two sets of primers specific for a wide range of dermatophytes were used to detect ribosomal DNA regions. The real-time PCR High Resolution Melting Analysis (PCR-HRMA) method was used for dermatophyte species identification. The PCR detection success was 74% for both sets of primers, the PCR-HRMA method enabled dermatophyte species identification in all PCR positive cases. In contrast, only 52% of patients with dermatophytosis were culture positive. Microscopy from the sample was positive in 90% of patients with proven dermatophytosis. 90% of dermatophytes were successfully identified using microscopy, culture and PCR-HRMA. The most common dermatophyte species (Trichophyton rubrum, T. interdigitale, T. benhamiae) were reliably detected by this methodology. PCR detection of rDNA directly from clinical material using HRMA increased the number of species identificacions in the diagnosis of dermatophytosis. The combination of classical and molecular biologic examinations appears to be a suitable method for the rapid and reliable diagnosis of dermatophytosis.

• Klíčová slova
• Trichophyton rubrum, Trichophyton interdigitale, Trichophyton benhamia,
• MeSH
• dermatomykózy * diagnóza   mikrobiologie   MeSH
• diagnostické techniky molekulární * metody   MeSH
• klinická studie jako téma MeSH
• lidé MeSH
• ribozomální DNA MeSH
• Trichophyton MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH