Rapid identification of filamentous fungi by colony PCR / 中华皮肤科杂志

Objective To estimate the application value of colony PCR in the detection of pathogenic filamentous fungi. Methods Colony PCR was established and performed to amplify the internal transcribed spacer (ITS) region of 19 species (strains) of filamentous fungus followed by sequencing analysis. At the same time, DNA extracts from 8 of the 19 species of filamentous fungus were subjected to conventional PCR. Hha I and Hinf I endonucleases were used for restriction fragment length polymorphism (RFLP) analysis of the conventional and colony PCR products. Comparison analysis was carried out between the colony and conventional PCR. Results Of the 19 strains, 16(84.2%) yielded positive results by colony PCR; sequence analysis of the PCR products of ITS region revealed a 96% - 100% similarity with the reference sequence (NCBI database)of corresponding fungi. The amplification product length and RFLP profile of these products from the 8 species of filamentous fungus, except for those from Aspergillus nidulans, were consistent between the colony and conventional PCR. Conclusions Compared with conventional PCR, colony PCR-based detection of filamentous fungi is easy to operate, time and labor-saving, with high accuracy and reliability, and can be applied to the rapid identification of filamentous fungi.

Clinical phenotype and pathogen profile of 7251 cases of cutaneous and mucous mycosis in Nanchang region / 中华皮肤科杂志

Objective To profile the phenotype and pathogens of cutaneous and mucous mycoses in a dermatology outpatient clinic in Nanchang region. Methods A review was performed to assess cutaneous and mucous mycoses diagnosed in the dermatology outpatient clinic of Dermatology Hospital of Jiangxi Province from 2006 to 2008. The relationship of clinical phenotype and pathogens to season, patients' age and gender was analyzed. Results A total of 7251 cases were collected, and the ratio of male to female patients was 2.3: 1. The most prevalent mycoses included tinea cruris (2702, 37.1%), pityriasis versicolor (1505, 20.8%) and tinea manus (727, 10.0%). In total, 4953 fungal strains were isolated from all the patients except for those with pityriasis versicolor, of them, Trichophyton rubrum accounted to 69.9%, Candida to 20.4%, and Trichophyton violaceum to 4.5%. Season, patients' age and gender were found to be associated with clinical phenotypes and pathogens of mycoses. Conclusions In the dermatology outpatient clinic of Nanchang region, tinea cruris is the most common superficial fungal disease, with the predominant pathogen being Trichophyton rubrum. Trichophyton violaceum is the primary pathogen of tinea capitis, which is different from other reports.

Isolation of Pathogenic Fungi from Patients with Onychomycosis by Multipoint Inoculation Method / 中华皮肤科杂志

Objectives To compare the difference between multipoint inoculation and routine method for isolation of pathogenic fungi from nail samples of onychomycosis,and to analyze the epidemiology of pathogenic fungi in those patients.Methods The nail clipping samples from each patient were inoculated onto the plates with Sabouraud's agar,Sabouraud's agar without cycloheximide and medium containing rapeseed oil,respectively,by an approach of at least seven inoculating points in each plate (multipoint inoculation),and onto medium slope in tubes with the same media as above mentioned (routine method).In the multipoint inoculation method,plates with more than 3 colonies were taken for further identification of pathogenic fungi based on morphological and biochemical properties.Results Based on the data from 150 samples of onychomycosis,significant differences were found between multipoint inoculation method and routine method (P

Studies on Susceptibility of Nine Antifungal Agents Against Dematiaceous Fungi by NCCLS M38-P Method / 中华皮肤科杂志

Objectives To evaluate the application of M38-P method recommend by NCCLS to dematiaceous fungi, and detect the susceptibility of itraconazole, fluconazole, miconazole, econazole, flucytosine, clotrimazole, nystatin, amphotericin B and terbinafine against dematiaceous fungi. Methods Using M38-P method forty-five strains of dematiaceous fungi were studied, which included 2 strains of Bipolaris spp., 1 strain of Drechslera spp., 1 strain of Curvularia spp.,14 strains of C. carrionii, 8 strains of F. pedrosoi, 3 strains of F. compacta and 16 strains of P. verrucosa. The media containing the fungi in the concentrations of (0.4 ~ 5) ? 106 CFU/mL were incubated at 35℃ for 5 ~ 7 days. Results The tested dematiaceous fungi were highly susceptible to itraconazole and terbinafine. Some of them were resistant to itraconzole, fluconazole, flucytosine and amphotericin B. Conclusions The data suggest that M38-P standards could be applied as the susceptibility testing of dematiaceous fungi, and it is necessary to monitor the resistant strains in antifungal treatment.