Species identification and strain differentiation of dermatophyte fungi by analysis of ribosomal-DNA intergenic spacer regions.
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Restriction fragment length polymorphisms (RFLPs) identified in the ribosomal-DNA (rDNA) repeat were used for molecular strain differentiation of the dermatophyte fungus Trichophyton rubrum. The polymorphisms were detected by hybridization of EcoRI-digested T. rubrum genomic DNAs with a probe amplified from the small-subunit (18S) rDNA and adjacent internal transcribed spacer (ITS) regions. The rDNA RFLPs mapped to the nontranscribed spacer (NTS) region of the rDNA repeat and appeared similar to those caused by short repetitive sequences in the intergenic spacers of other fungi. Fourteen individual RFLP patterns (DNA types A to N) were recognized among 50 random clinical isolates of T. rubrum. A majority of strains (19 of 50 [38%]) were characterized by one RFLP pattern (DNA type A), and four types (DNA types A to D) accounted for 78% (39 of 50) of all strains. The remaining types (DNA types E to N) were represented by one or two isolates only. A rapid and simple method was also developed for molecular species identification of dermatophyte fungi. The contiguous ITS and 5.8S rDNA regions were amplified from 17 common dermatophyte species by using the universal primers ITS 1 and ITS 4. Digestion of the amplified ITS products with the restriction endonuclease MvaI produced unique and easily identifiable fragment patterns for a majority of species. However, some closely related taxon pairs, such as T. rubrum-T. soudanense and T. quinkeanum-T. schoenlenii could not be distinguished. We conclude that RFLP analysis of the NTS and ITS intergenic regions of the rDNA repeat is a valuable technique both for molecular strain differentiation of T. rubrum and for species identification of common dermatophyte fungi. (+info)
Systematic review of topical treatments for fungal infections of the skin and nails of the feet.
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OBJECTIVE: To identify and synthesise the evidence for efficacy and cost effectiveness of topical treatments for superficial fungal infections of the skin and nails of the feet. DESIGN: Systematic review. INTERVENTIONS: Topical treatments for superficial fungal infections. MAIN OUTCOME MEASURES: Cure confirmed by culture and microscopy for skin and by culture for nails in patients with clinically diagnosed fungal infections. RESULTS: Of 126 trials identified in 121 papers, 72 (57.1%) met the inclusion criteria. Placebo controlled trials yielded pooled relative risks of failure to cure skin infections: allylamines (0.30, 95% confidence interval 0.24 to 0.38); azoles (0.54, 0.42 to 0.68); undecenoic acid (0.28, 0. 11 to 0.74); and tolnaftate (0.46, 0.17 to 1.22). Although meta-analysis of 11 trials comparing allylamines and azoles showed a relative risk of failure to cure of 0.88 (0.78 to 0.99) in favour of allylamines, there was evidence of language bias. Seven reports in English favoured allylamines (0.79, 0.69 to 0.91), but four reports in foreign languages showed no difference between the two drugs (1. 01, 0.90 to 1.13). Neither trial of nail infections showed significant differences between alternative topical treatments. CONCLUSIONS: Allylamines, azoles, and undecenoic acid were efficacious in placebo controlled trials. There are sufficient comparative trials to judge relative efficacy only between allylamines and azoles. Allylamines cure slightly more infections than azoles but are much more expensive than azoles. The most cost effective strategy is first to treat with azoles or undecenoic acid and to use allylamines only if that fails. (+info)
Genetic diversity in the yeast species Malassezia pachydermatis analysed by multilocus enzyme electrophoresis.
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Fifty-two strains of the yeast species Malassezia pachydermatis were analysed by multilocus enzyme electrophoresis. M. pachydermatis appeared to be genetically heterogeneous. A total of 27 electrophoretic types were identified that could be divided into five distinct groups with different host specificities. The diversity revealed by this electrophoretic method matched remarkably well the reported genetic variability obtained by comparing large subunit rRNA sequences. This study also suggests that genetic exchanges can occur in the anamorphic species M. pachydermatis. (+info)
Endemic regions of paracoccidioidomycosis in Brazil: a clinical and epidemiologic study of 584 cases in the southeast region.
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This paper describes the clinical-seroepidemiologic characteristics of patients with paracoccidioidomycosis (PCM) who visited the University Hospital at the State University of Campinas (Campinas, Sao Paulo, Brazil). The study group consisted of 584 individuals (492 males and 92 females) with ages ranging from 5 to 87 years. The highest incidence of the disease occurred between the ages of 41 and 50 years for men and between 11 and 40 years for women. Rural activities were the principal occupation of 46% of the patients. The diagnosis was confirmed by histopathologic examination and demonstration of fungus in scrapings, secretions, or in the sputum. Serologic test results for PCM were positive in 80% of the 584 patients studied. The significant number of patients, including 33 children less than 14 years old, indicates the presence of the fungus in the area and that this region is an important endemic area for PCM. (+info)
Aspergillosis in children with cancer: A 34-year experience.
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A retrospective review of medical records, microbiology and pathology laboratory records, and nosocomial infection surveillance data was undertaken to describe the experience with culture-documented aspergillus infection in pediatric cancer patients at our facility. Sixty-six patients were identified from a 34-year period. The most common underlying diagnosis was leukemia. Risk factors included neutropenia, immunosuppression, and prior antibiotic therapy. On the basis of clinical presentation, 23 patients were believed to have disseminated disease and 43 to have localized disease. The lung was the most frequently affected organ. Despite aggressive medical and surgical management, overall mortality was 85% within the first year after diagnosis. Patients who presented with disease in sites other than the lungs fared better than patients with initial pulmonary involvement (P=.0014). Aspergillosis continues to be associated with poor outcome. Development of improved medical and adjuvant therapies, including surgery, is warranted. (+info)
Reverse transcription - 3' rapid amplification of cDNA ends-nested PCR of ACT1 and SAP2 mRNA as a means of detecting viable Candida albicans in an in vitro cutaneous candidiasis model.
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The presence of viable cells of Candida albicans, in broth or in a reconstructed living skin equivalent, was determined by the detection of amplicons of partial mRNA sequences of the genes encoding fungal actin (ACT1) and secreted aspartyl proteinase 2 (SAP2). The mRNA of both genes were amplified by reverse transcription-3' rapid amplification of cDNA ends-nested polymerase chain reaction. Single bands of ACT1 (315 bp) and SAP2 (162 bp) mRNA were amplified from total RNA extracts of C. albicans grown in yeast carbon base-albumin broth or in living skin equivalent tissue; only the former was amplified from Sabouraud broth-grown organisms. Primer pairs targeted for ACT1 and SAP2 were Candida genus-specific and C. albicans-specific, respectively. The sensitivity limits of the assay were 100 fg of total RNA or 10 cells of C. albicans, by ethidium bromide staining. When C. albicans-infected living skin equivalent was exposed to amorolfine, amplicons of ACT1 and SAP2 mRNA were not detected in total RNA extracts. Non-amplification of the mRNA correlated with the absence of C. albicans growth in Sabouraud agar cultures of living skin equivalent samples. Reverse transcription-3' rapid amplification of cDNA ends-nested polymerase chain reaction of the mRNA encoding specific proteins of an organism has potential application in determining the viability of the organism in tissue, thus monitoring the efficacy of an antimicrobial therapy, and in detecting mRNA expressed in very little amounts in tissue. (+info)
Diagnosing dermatomycosis in general practice.
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BACKGROUND: Diagnosing dermatomycosis from a clinical image is not always easy. Microscopy of a potassium hydroxide preparation (KOH-test) and culturing are seldomly used in general practice. Cyanoacrylate surface skin scraping (CSSS) is a new diagnostic tool that may be useful and simple. OBJECTIVES: We aimed to investigate the diagnostic value of signs and symptoms, the KOH-test and the CSSS, in patients with erythematosquamous skin lesions, using the culture as the gold standard. Our goal is to formulate an optimal algorithm for the diagnosis of mycosis, based on one or more of these tests and including both optimal accuracy and costs. METHODS: Scales from 148 consecutive general practice patients were tested using a KOH-test, CSSS and culture. Clinical data were collected using a questionnaire. RESULTS: Twenty-six (18%) positive fungal cultures were identified. The sensitivity of the clinical diagnosis was 81% and its specificity 45%; for the KOH-test, these figures were 12 and 93% respectively; and for the CSSS, 62 and 88%, respectively. The positive predictive value of the clinical diagnosis was 24% and the negative predictive value 92%; for the KOH-test these figures were 25 and 83%, respectively, and for the CSSS, 52 and 92%, respectively. Determining CSSS in all patients proved to be the most accurate policy (accuracy = 83%). The likelihood ratio of CSSS in all patients was 5.17 for a positive test result and 0.43 for a negative test result. An approach in which CSSS is obtained in only those patients whom the physician considers by clinical examination to have dermatomycosis, with no testing in other patients, results in positive and negative likelihood ratios of 4.69 and 0.56, respectively. Such a policy would result in an overall sensitivity of 50%, a specificity of 89%, a positive predictive value of 50% and a negative predictive value of 89%. DISCUSSION: The clinical picture of dermatomycosis is not very reliable. The combination of a clinical judgement if this is negative and an additional CSSS in the case of a positive clinical judgement provides us with the best cost-benefit ratio, if both diagnostic accuracy and logistic considerations are taken into consideration. (+info)
Species identification and strain typing of Malassezia species stock strains and clinical isolates based on the DNA sequences of nuclear ribosomal internal transcribed spacer 1 regions.
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This study demonstrated the application of internal transcribed spacer 1 (ITS1) ribosomal DNA sequences to the species identification and strain typing of 28 standard strains and 46 clinical isolates of the genus Malassezia. The size of ITS1 regions ranged from 162 to 266 bp. Members of the genus Malassezia (M. pachydermatis, M. furfur, M. sympodialis, M. globosa, M. obtusa, M. restricta and M. slooffiae) were classified into seven ITS1-homologous groups and 22 ITS1-identical, individual groups. The 46 clinical isolates of lipophilic Malassezia spp. were identified as belonging to just three ITS1-homologous groups, i.e., M. furfur (19 strains: 11 from pityriasis versicolor, 4 from seborrhoeic dermatitis and 4 from atopic dermatitis). M. sympodialis (22 strains: 7 from pityriasis versicolor, 3 from seborrhoeic dermatitis, 1 from atopic dermatitis and 11 from healthy controls) and M. slooffiae (five strains: three from chronic otitis media and two from healthy controls). (+info)