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1.
J Pharmacol Sci ; 144(1): 52-56, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32565006

RESUMO

The use of biological drugs in psoriasis is replacing traditional therapies due to their specific mechanism and limited side effects. However, the use of Interleukin 17 inhibitors and the modification of its cytokine pathway could favor the risk of fungal infections. All-trans retinoic acid is an active metabolite of vitamin A with anti-inflammatory and immunoregulatory properties through its capacity to stimulate both innate and adaptive immunity and to its effects on proliferation, differentiation and apoptosis in a variety of immune cells. Furthermore, it has been recently discovered that All-trans retinoic acid has a direct fungistatic effect against Candida and Aspergillus Fumigatus. On the basis of these new insights, in the current review, we suggest that the evaluation of serum level of All-trans retinoic acid or vitamin A should be considered as a predictive marker for the development of fungal infections among psoriatic patients treated with Interleukin 17 inhibitors. In clinical practice, vitamin A test could be added in the routine hospital diagnostic management for a better selection of psoriatic patients eligible to Interleukin 17 inhibitors.


Assuntos
Anticorpos Monoclonais Humanizados/efeitos adversos , Candidíase/diagnóstico , Candidíase/etiologia , Dermatomicoses/diagnóstico , Dermatomicoses/etiologia , Interleucina-17/antagonistas & inibidores , Micoses/diagnóstico , Micoses/etiologia , Psoríase/tratamento farmacológico , Psoríase/imunologia , Vitamina A/sangue , Biomarcadores/sangue , Candidíase/prevenção & controle , Citocinas/metabolismo , Dermatomicoses/prevenção & controle , Humanos , Interleucina-17/metabolismo , Micoses/prevenção & controle , Seleção de Pacientes , Valor Preditivo dos Testes , Risco , Transdução de Sinais/efeitos dos fármacos , Tretinoína/sangue
2.
Artigo em Inglês | MEDLINE | ID: mdl-32578682

RESUMO

Dermatophytes are known as a common cause of superficial mycosis, but atypical presentations in immunosuppressed patients make the diagnosis more challenging. Here, we report a case of a 39-year-old patient, a renal transplant recipient from a living donor, who presented with atypical cutaneous lesions of lower extremities caused by Arthroderma gypseum (Nannizzia gypsea), four months after receiving a renal transplant. It is important to highlight the importance of the early detection of fungal infections in immunosuppressed patients. Clinicians should have a high degree of suspicion for the early detection and treatment of the cases.


Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Transplante de Rim/efeitos adversos , Adulto , Dermatomicoses/patologia , Diagnóstico Diferencial , Feminino , Humanos , Hialoifomicose/diagnóstico , Hospedeiro Imunocomprometido
3.
Dermatol Ther ; 33(4): e13481, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32369240

RESUMO

In view of the new viral COVID-19 pandemic, the fungal Candida auris epidemic still in progress worldwide highlights non-Candida albicans candidal infections. We describe an immunocompetent woman with a cutaneous manifestation of Candida parasilopsis fungemia, a prominent eschar, which proved to be the nidus for the candidemia. We stress the value of selectively removing eschars. C. parasilopsis and C. auris are increasingly important causes of sepsis and wound infections. We emphasize that commercially available biochemical-based tests may misidentify C. auris as C. parapsilosis, and stress the added danger of C. auris to critically ill-hospitalized COVID-19 patients. Any health care facility with evidence of infection or colonization with C. auris requires very close monitoring, since this fungus is a nosocomial threat comparable to SARS-CoV-2 in its mortality and fomite adhesiveness! Both organisms have the potential to be transmitted as nosocomial pathogens; health care workers need to follow strict CDC guidelines. During this COVID-19 pandemic, every health care facility should closely monitor for the possible deadly combination of the SARS-CoV-2 and C. auris. The identification of C. auris necessitates use of sophisticated technology not readily available to make this essential diagnosis since C. auris is multi-drug resistant and isolation precautions would become paramount.


Assuntos
Betacoronavirus , Candida/isolamento & purificação , Candidemia/epidemiologia , Infecções por Coronavirus/epidemiologia , Dermatomicoses/epidemiologia , Pandemias , Pneumonia Viral/epidemiologia , Pele/microbiologia , Candidemia/microbiologia , Comorbidade , Dermatomicoses/diagnóstico , Dermatomicoses/microbiologia , Feminino , Humanos , Pessoa de Meia-Idade , Pele/patologia
4.
Infection ; 48(3): 429-434, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32232786

RESUMO

BACKGROUND: Fungal infections of the skin, hair, and nails are the largest and most widespread group of all mycoses. Nannizzia nana is a relatively rare etiological factor of dermatomycosis in humans, as it usually affects animals, e.g. pigs and boars. In addition to the zoophilic nature, there are also reports of the geophilic reservoir of this dermatophyte species. OBJECTIVE: In this study, we present symptomatic infections with N. nana aetiology in humans reported recently in Poland. Interestingly, these cases had a non-specific clinical picture and occurred as skin lesions on the neck and foot as well as onychomycosis of the toenails. From the medical history, the patients had no contact with pigs. METHODS: Diagnostics of these infections was performed with a combination of classical phenotypic and molecular genomic methods. The genomic diversity of the isolates was determined using the MP-PCR method. In vitro antifungal susceptibility tests against itraconazole, ketoconazole, terbinafine and naftifine hydrochloride were also performed. RESULTS: Nannizzia nana has been identified as an etiological factor of dermatomycosis. Moreover, heterogeneity of the genomes was revealed for the obtained strains. In vitro activities of antifungal agents showed that isolates were susceptible to all tested drugs. The patients were treated with oral terbinafine and topical ketoconazole cream, which led to a complete recovery. CONCLUSIONS: In conclusion, the cases studied by us may indicate that the infrequency of N. nana infections may not necessarily be related to the low infectivity of this fungal agent, but they are rather associated with misdiagnosis. Furthermore, N. nana reservoirs should also be sought in soil.


Assuntos
Antifúngicos/uso terapêutico , Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Cetoconazol/uso terapêutico , Administração Oral , Administração Tópica , Adulto , Idoso , Arthrodermataceae/classificação , Dermatomicoses/tratamento farmacológico , Dermatomicoses/microbiologia , Diagnóstico Diferencial , Feminino , Humanos , Masculino , Onicomicose/diagnóstico , Onicomicose/tratamento farmacológico , Onicomicose/microbiologia , Microbiologia do Solo , Terbinafina/uso terapêutico , Resultado do Tratamento
5.
J Appl Microbiol ; 129(2): 212-232, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32048417

RESUMO

Dermatophytes are the aetiological factors of a majority of superficial fungal infections. What distinguishes them from other pathogenic filamentous fungi is their unique ability to degrade keratin. The remarkable ability of this group of fungi to survive in different ecosystems results from their morphological and ecological diversity as well as high adaptability to changing environmental conditions. Paradoxically, despite the progress in medicine, the prevalence of dermatophyte infections is increasing from year to year. At the beginning of the third millennium, practical diagnostic and therapeutic options are still very limited. This review focuses on understanding the major problems in this aspect of dermatophyte infections and indicates future strategies and perspectives for novel approaches to identification and drugs for elimination of dermatophytes. Particular importance is placed on development of a strategy for a diagnostic pathway and implementation of rapid and reliable diagnostics methods designed by international teams. Furthermore, among compounds that currently arouse great interest, representatives of terpenoids, alkaloids, saponins, flavonoids and essential oils deserve attention. Many of these compounds are undergoing clinical trials as potential antifungal agents, and future research should focus on attempts at determination of the applicability of tested substances. Finally, the advantages and disadvantages in implementation of new diagnostic paths and medicinal substances for routine use are indicated.


Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , Antifúngicos/uso terapêutico , Arthrodermataceae/efeitos dos fármacos , Dermatomicoses/microbiologia , Desenvolvimento de Medicamentos , Ecossistema , Humanos , Técnicas Microbiológicas
6.
BMJ Case Rep ; 13(2)2020 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-32060111

RESUMO

Primary cutaneous aspergillosis (PCA) occurs through inoculation of fungal spores directly into the skin from the environment through disrupted skin such as in burns, surgery or penetrating trauma patients. Most cases reported in literature were in the immunocompromised, rarely in immunocompetent patients. The characteristic lesion of cutaneous aspergillosis is a black eschar on a red plaque, or nodule at the site of skin injury. The diagnosis of PCA can be made by identifying hyphal forms on routine H&E staining or special stains such as periodic acid-Schiff or Gomori methenamine-silver stains on skin biopsy and by fungal cultures. We report a case of an 80-year-old farmer who developed cutaneous aspergillosis after a surgical procedure without any systemic spread. The diagnosis was made by histopathology and tissue fungal cultures. He was treated with incision and drainage followed by oral voriconazole for 4 weeks; which led to clinical recovery.


Assuntos
Aspergilose/diagnóstico , Aspergillus fumigatus/isolamento & purificação , Dermatomicoses/diagnóstico , Idoso de 80 Anos ou mais , Aspergilose/tratamento farmacológico , Dermatomicoses/tratamento farmacológico , Humanos , Imunocompetência , Masculino , Complicações Pós-Operatórias , Coloração e Rotulagem , Voriconazol/uso terapêutico
7.
BMC Infect Dis ; 20(1): 171, 2020 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-32087692

RESUMO

BACKGROUND: Trichophyton benhamiae is a zoophilic dermatophyte that can cause tinea in humans and animals. Lesions caused by T. benhamiae tend to be highly inflammatory, and patients are often infected by animals or other patients infected with T. benhamiae. In this paper, we report the first case of tinea faciei caused by T. benhamiae in a Chinese girl who might be transmitted from a fox. CASE PRESENTATION: A 4-year-old girl from HaiNing city developed an itchy, erythematous, and annular plaque on her right face for the past 2 months. Before the lesion appeared, she was in close contact with the fur of a fox for almost 1 week. Septate hyaline hyphae were detected by direct mycological examination of the scales. Cultures grew on Sabouraud's dextrose agar (SDA) at 26 °C for 2 weeks revealed the presence of T. mentagrophytes. A molecular sequencing test confirmed that the isolate was consistent with reference strains to T. benhamiae. Then, the diagnosis of tinea faciei due to T. benhamiae was made. Treatment with terbinafine (oral 125 mg/d) and sertaconazole nitrate cream (topical, twice daily) for 4 weeks was initiated and achieved significant improvement of the skin lesions. CONCLUSIONS: This rare dermatophytosis case highlights the importance of ITS sequencing in helping to recognize rare pathogenic fungi that can be easily misdiagnosed with a conventional morphological diagnosis.


Assuntos
Arthrodermataceae/genética , Dermatomicoses/diagnóstico , Tinha/diagnóstico , Trichophyton/genética , Administração Oral , Administração Tópica , Animais , Antifúngicos/administração & dosagem , Antifúngicos/uso terapêutico , Arthrodermataceae/isolamento & purificação , Pré-Escolar , China , Dermatomicoses/tratamento farmacológico , Dermatomicoses/microbiologia , Face/patologia , Feminino , Humanos , Imidazóis/administração & dosagem , Imidazóis/uso terapêutico , Pele/patologia , Terbinafina/administração & dosagem , Terbinafina/uso terapêutico , Tiofenos/administração & dosagem , Tiofenos/uso terapêutico , Tinha/tratamento farmacológico , Tinha/microbiologia , Resultado do Tratamento , Trichophyton/isolamento & purificação
8.
Vet Dermatol ; 31(1): 28-74, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31957204

RESUMO

BACKGROUND: The genus Malassezia is comprised of a group of lipophilic yeasts that have evolved as skin commensals and opportunistic cutaneous pathogens of a variety of mammals and birds. OBJECTIVES: The objective of this document is to provide the veterinary community and other interested parties with current information on the ecology, pathophysiology, diagnosis, treatment and prevention of skin diseases associated with Malassezia yeasts in dogs and cats. METHODS AND MATERIAL: The authors served as a Guideline Panel (GP) and reviewed the literature available prior to October 2018. The GP prepared a detailed literature review and made recommendations on selected topics. The World Association of Veterinary Dermatology (WAVD) Clinical Consensus Guideline committee provided guidance and oversight for this process. The document was presented at two international meetings of veterinary dermatology societies and one international mycology workshop; it was made available for comment on the WAVD website for a period of six months. Comments were shared with the GP electronically and responses incorporated into the final document. CONCLUSIONS AND CLINICAL IMPORTANCE: There has been a remarkable expansion of knowledge on Malassezia yeasts and their role in animal disease, particularly since the early 1990's. Malassezia dermatitis in dogs and cats has evolved from a disease of obscurity and controversy on its existence, to now being a routine diagnosis in general veterinary practice. Clinical signs are well recognised and diagnostic approaches are well developed. A range of topical and systemic therapies is known to be effective, especially when predisposing factors are identified and corrected.


Assuntos
Doenças do Gato/diagnóstico , Doenças do Gato/tratamento farmacológico , Dermatite/veterinária , Dermatomicoses/veterinária , Doenças do Cão/diagnóstico , Doenças do Cão/tratamento farmacológico , Animais , Antifúngicos/uso terapêutico , Gatos , Consenso , Dermatite/diagnóstico , Dermatite/tratamento farmacológico , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , Cães , Malassezia/efeitos dos fármacos , Pele/microbiologia , Pele/patologia , Medicina Veterinária/métodos , Medicina Veterinária/organização & administração
10.
Diagn Cytopathol ; 48(4): 401-404, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31930706

RESUMO

A young woman on long term corticosteroid therapy presented with a discrete hyperpigmented nodular cystic swelling on her upper thigh to the buttock region. There was history of discharging sinuses however at presentation the lesion was painless nondischarging. The lesion was subjected to fine-needle aspiration. The cytological smears on routine and special stains showed branching, septate fungal hyphae present extracellularly and intracellularly. The background showed dense mixed inflammation and granulomatous inflammation. The aspirated material was further subjected to culture. A nonsporulating melanized fungus was obtained and a culture isolate was sent for molecular characterization. Medicopsis romeroi, a rare melanized fungus belonging to the order Pleosporales was isolated on Internal transcribed spacer sequencing.


Assuntos
Ascomicetos/genética , Dermatomicoses , Feoifomicose , Adulto , Ascomicetos/classificação , Biópsia por Agulha Fina , Dermatomicoses/diagnóstico , Dermatomicoses/genética , Dermatomicoses/microbiologia , Feminino , Humanos , Feoifomicose/diagnóstico , Feoifomicose/genética , Feoifomicose/microbiologia
11.
FP Essent ; 489: 16-20, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31995350

RESUMO

Fungal and viral skin infections are common and typically are managed by family physicians. The fungal skin infections commonly seen in family practice include the various forms of tinea: tinea corporis, tinea gladiatorum, tinea cruris, tinea pedis, tinea capitis, and tinea unguium (eg, onychomycosis). Common viral skin infections include herpes simplex virus infection, herpes zoster, cutaneous and genital warts, and molluscum contagiosum. Many fungal and viral skin infections have a classic appearance but variations and atypical manifestations can make patients with these conditions difficult to diagnose definitively. Confirmatory testing often is not required. In complex cases with atypical features, microscopy, culture, or blood tests can help in making a diagnosis. For some infections, treatment may be initiated before confirmatory test results are received. Most fungal skin infections can be managed effectively with topical antifungals. Tinea capitis and onychomycosis should be managed with oral drugs. Oral antiviral drugs are used to manage most viral skin infections but dosages vary based on the condition and phase of the infectious process. Cutaneous warts typically are managed with salicylic acid and/or cryotherapy.


Assuntos
Queimaduras , Dermatomicoses , Dermatopatias Infecciosas , Tinha , Queimaduras/complicações , Queimaduras/microbiologia , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , Dermatomicoses/etiologia , Humanos , Pacientes Ambulatoriais , Dermatopatias Infecciosas/diagnóstico , Dermatopatias Infecciosas/tratamento farmacológico , Dermatopatias Infecciosas/etiologia , Tinha dos Pés
12.
Med Mycol J ; 60(4): 101-107, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31787730

RESUMO

Cutaneous cryptococcosis is classified either as primary or secondary based on the route of infection. The disease can also be classified either as localized cutaneous cryptococcosis or cutaneous manifestations of disseminated cryptococcosis. However, from a physician's point of view, whether lesions are localized to the skin or are disseminated/systemic is more important than the route of infection. The Clinical Practice Guidelines for Diagnosis and Treatment of Cryptococcosis, which was established in 2019 by the Japanese Society for Medical Mycology, adopted the latter classification. Localized cutaneous cryptococcosis is defined as a condition in which lesions are confined within a limited part of the skin, not systemically disseminated at the same time, and are associated with neither cryptococcal fungemia nor antigenemia. This type of cutaneous cryptococcosis is uncommon in Japan. Only 65 cases were reported during the 50-year study period from 1968 to August 2018, with the patients divided into two groups: immunocompromised patients (n=44, 68%) and immunocompetent patients (n=21, 32%). None of the patients were infected with the human immunodeficiency virus (HIV). Localized cutaneous cryptococcosis can also occur in non-HIV-infected patients and well-appearing individuals, therefore, it is considered an important infection in routine dermatology practice. Here, we outline the classification, diagnosis, and treatment of cutaneous cryptococcosis and present a summary of cutaneous cryptococcosis cases reported in Japan.


Assuntos
Criptococose , Dermatomicoses , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Criptococose/classificação , Criptococose/diagnóstico , Criptococose/tratamento farmacológico , Criptococose/patologia , Dermatomicoses/classificação , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , Dermatomicoses/patologia , Feminino , Humanos , Imunocompetência , Hospedeiro Imunocomprometido , Masculino , Pessoa de Meia-Idade
16.
Mycopathologia ; 184(5): 691-697, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31606812

RESUMO

Corynespora cassiicola is a common plant pathogen, but C. cassiicola infection in human hosts is extremely rare. In this report, we present an 84-year-old male with long-term use of inhaled corticosteroids who developed a subcutaneous infection caused by C. cassiicola. The organism was isolated from both wound culture and biopsy specimen from the skin lesion. However, no microscopic diagnostic characters could be obtained because the isolates failed to sporulate on different culture media. Molecular diagnosis by amplification and sequencing of the internal transcribed spacer regions of ribosomal DNA was performed, and the sequences of the isolates were identical to those of C. cassiicola. The patient was treated successfully with oral terbinafine therapy for 12 weeks. In this report, we also review the epidemiology, clinical and therapeutic facets of cutaneous C. cassiicola infection.


Assuntos
Antifúngicos/uso terapêutico , Ascomicetos/isolamento & purificação , Dermatomicoses , Infecções Oportunistas/diagnóstico , Terbinafina/uso terapêutico , Administração por Inalação , Corticosteroides/administração & dosagem , Corticosteroides/efeitos adversos , Idoso de 80 Anos ou mais , Ascomicetos/classificação , Dermatomicoses/diagnóstico , Dermatomicoses/tratamento farmacológico , Dermatomicoses/epidemiologia , Humanos , Masculino , Infecções Oportunistas/tratamento farmacológico
17.
Dermatol Online J ; 25(8)2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31553862

RESUMO

Histoplasmosis is a systemic mycosis caused by the dimorphic fungus Histoplasma capsulatum, with disseminated histoplasmosis (HD) being one of its clinical forms. As a consequence of the HIV-AIDS pandemic, HD has become prevalent not only in regions that are recognized as endemic but also in areas not considered endemic, such as Europe and Asia. Its clinical manifestations are varied and mimic several infectious diseases, mainly tuberculosis. In endemic areas, it is the first manifestation of AIDS in 50 to 70% of patients. The diagnosis of histoplasmosis is difficult and HD can lead to death if not diagnosed early and if proper treatment is not instituted. The present report presents a patient with a recent diagnosis of HIV-AIDS, in treatment for miliary tuberculosis, who was diagnosed with disseminated histoplasmosis because of his dermatological manifestations.


Assuntos
Síndrome de Imunodeficiência Adquirida/tratamento farmacológico , Fármacos Anti-HIV/uso terapêutico , Antituberculosos/uso terapêutico , Dermatomicoses/diagnóstico , Histoplasmose/diagnóstico , Tuberculose Miliar/tratamento farmacológico , Síndrome de Imunodeficiência Adquirida/complicações , Anfotericina B/uso terapêutico , Antifúngicos/uso terapêutico , Estado Terminal , Dermatomicoses/complicações , Dermatomicoses/tratamento farmacológico , Dermatomicoses/patologia , Histoplasma , Histoplasmose/complicações , Histoplasmose/tratamento farmacológico , Histoplasmose/patologia , Humanos , Itraconazol/uso terapêutico , Masculino , Tuberculose Miliar/complicações , Adulto Jovem
18.
Mycoses ; 62(12): 1116-1126, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31493312

RESUMO

BACKGROUND: Since accurate identification of dermatophyte species is essential for epidemiological studies and implementing antifungal treatment, overcoming limitations of conventional diagnostics is a fruitful subject. OBJECTIVES AND METHODS: In this study, we investigated real-time polymerase chain reaction(q-PCR), matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) and nano-electrospray ionisation mass spectrometry (nano-ESI-MS) to detect and identify the most frequently isolated dermatophytes from human and animal dermatophytosis in comparison with conventional methods. RESULTS: Among 200 samples, the identified species were Microsporum canis (78.22%), Trichophyton verrucosum (10.89%) and T. mentagrophytes (5.94%). Q-PCR assay displayed great execution attributes for dermatophytes detection and identification. Using MALDI-TOF MS, M. canis, but none of T. violacium, T. verrucosum or T. mentagrophytes, could be identified. Nano-ESI-MS accurately identified all species. The potential virulence attributes of secreted proteases were anticipated and compared between species. Secreted endoproteases belonging to families/subfamilies of metalloproteases, subtilisins and aspartic protease were detected. The analysed exoproteases are aminopeptidases, dipeptidyl peptidases and carboxypeptidases. Microsporum canis have three immunogenic proteins, siderophore iron transporter mirB, protease inhibitors, plasma membrane proteolipid 3 and annexin. CONCLUSION: In essence, q-PCR, MALDI-TOF MS and nano-ESI-MS assays are very nearly defeating difficulties of dermatophytes detection and identification, thereby, supplement or supplant conventional diagnosis of dermatophytosis.


Assuntos
Arthrodermataceae/classificação , Dermatomicoses/microbiologia , Proteômica , Adolescente , Adulto , Animais , Arthrodermataceae/química , Gatos/microbiologia , Bovinos/microbiologia , Criança , DNA Fúngico/isolamento & purificação , Dermatomicoses/diagnóstico , Cães/microbiologia , Feminino , Cavalos/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Adulto Jovem
19.
J Med Microbiol ; 68(11): 1641-1648, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31526456

RESUMO

Introduction: Onychomycosis is a debilitating, difficult-to-treat nail fungal infection with increasing prevalence worldwide. The main etiological agents are dermatophytes, which are common causative pathogens in superficial fungal mycoses. Conventional detection methods such as fungal culture have low sensitivity and specificity and are time-consuming.Aim: The main objective of this study was to design, develop and validate a real-time probe-based multiplex qPCR assay for the detection of dermatophytes and Fusarium species.Methodology: The performance characteristics of the qPCR assays were evaluated. The multiplex qPCR assays targeted four genes (assay 1: pan-dermatophytes/Fusarium spp.; assay 2: Trichophyton rubrum/Microsporum spp.). Analytical validation was accomplished using 150 fungal isolates and clinical validation was done on 204 nail specimens. The performance parameters were compared against the gold standard (fungal culture) and expanded gold standard (culture in conjunction with sequencing).Results: Both the single-plex and multiplex qPCR assays performed well especially when compared against the expanded gold standard. Among the 204 tested nail specimens, the culture method showed that 125 (61.3 %) were infected with at least one organism, of which 40 yielded positive results for dermatophytes and Fusarium spp. These target organisms detected include 20 dermatophytes and 22 Fusarium spp. The developed qPCR assays demonstrated excellent limit of detection, efficiency, coefficient of determination, analytical and clinical sensitivity and specificity.Conclusion: The multiplex qPCR assays were reliable for the diagnosis of onychomycosis, with shorter turn-around time as compared to culture method. This aids in the planning of treatment strategies to achieve optimal therapeutic outcome.


Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/microbiologia , Fusariose/microbiologia , Fusarium/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Arthrodermataceae/classificação , Arthrodermataceae/genética , Dermatomicoses/diagnóstico , Fusariose/diagnóstico , Fusarium/genética , Humanos , Sensibilidade e Especificidade
20.
PLoS One ; 14(9): e0222360, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31513657

RESUMO

Precise knowledge of the health status of experimental fish is crucial to obtain high scientific and ethical standards in biomedical research. In addition to the use of sentinel fish, the examination of diseased fish is a fundamental part of all health monitoring concepts. PCR assays offer excellent sensitivity and the ability to test a broad variety of pathogenic agents in different sample types. Recently, it was shown that analysis of environmental samples such as water, sludge or detritus from static tanks can complement PCR analysis of fish and is actually more reliable for certain pathogens. In our study, we investigated whether the analysis of filtered water mixed with detritus of tanks including fish showing clinical signs of illness is suitable to complement health monitoring programs in recirculating systems. The obtained data indicate that pathogens such as Pseudoloma neurophilia or Myxidium streisingeri were exclusively or mainly found in fish, while mycobacteria were predominantly present in environmental samples. A combination of both sample types seems to be required for the detection of a broad range of infectious agents in zebrafish colonies using real-time PCR technology.


Assuntos
Doenças dos Peixes/diagnóstico , Peixe-Zebra/microbiologia , Peixe-Zebra/parasitologia , Animais , Bactérias/patogenicidade , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/veterinária , Dermatomicoses/diagnóstico , Dermatomicoses/veterinária , Doenças dos Peixes/microbiologia , Doenças dos Peixes/parasitologia , Laboratórios , Microsporidiose/parasitologia , Mycobacterium/patogenicidade , Infecções por Mycobacterium/diagnóstico , Infecções por Mycobacterium/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Vigilância Epidemiológica Baseada em Águas Residuárias , Água/análise
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