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).
- Publication type
- Journal Article 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 isolation & purification MeSH
- Humans MeSH
- Mycological Typing Techniques methods MeSH
- Onychomycosis * therapy MeSH
- Plasma Gases therapeutic use MeSH
- In Vitro Techniques MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
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
- Brain Abscess diagnostic imaging microbiology surgery MeSH
- Amphotericin B therapeutic use MeSH
- Antifungal Agents pharmacology therapeutic use MeSH
- Ascomycota drug effects genetics isolation & purification MeSH
- Immunocompetence MeSH
- Middle Aged MeSH
- Humans MeSH
- Mycoses diagnosis diagnostic imaging MeSH
- DNA, Ribosomal genetics MeSH
- Treatment Outcome MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Case Reports MeSH
- Review MeSH
- Geographicals
- Czech Republic MeSH
- MeSH
- Congresses as Topic MeSH
- Humans MeSH
- Health Promotion * MeSH
- Primary Prevention * MeSH
- Primary Health Care MeSH
- Preventive Health Services MeSH
- Check Tag
- Humans MeSH
- Publication type
- News 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.
- Keywords
- Trichophyton rubrum, Trichophyton interdigitale, Trichophyton benhamia,
- MeSH
- Dermatomycoses * diagnosis microbiology MeSH
- Molecular Diagnostic Techniques * methods MeSH
- Clinical Studies as Topic MeSH
- Humans MeSH
- DNA, Ribosomal MeSH
- Trichophyton MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
