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Iranian Journal of Public Health، جلد ۳۸، شماره ۱، صفحات ۱۸-۲۴
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عنوان انگلیسی Application of PCR-RFLP to Rapid Identification of the Main Pathogenic Dermatophytes from Clinical Specimens
چکیده انگلیسی مقاله Background: In the present study, a PCR-RFLP based molecular technique was designed to rapid identification of der­matophytes in clinical specimens. Skin scrapings obtained from human cases suspected to dermatophytosis were studied in or­der to identify involved etiological fungi. Methods: In this experimental study, the specimens (skin scrapings) of patients referred to Mycology Department of Pas­teur Institute of Iran were inoculated on Petri dishes contained selective agar for pathogenic fungi (SAPF) and incubated at 25º C until visible growth of fungal colonies. The colonies were examined for standard morphological characteristics after visi­ble growth on the agar medium. A small portion of each fungal colony was further studied by restriction fragment length poly­morphism (RFLP) analysis of the PCR-amplified internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). PCR amplicons were electrophoresed on 2% agarose gel after digesting by different restriction enzymes including Mva I, Hinf I and Hae III. Results: Among 160 clinical samples examined, 6 dermatophyte species including Trichophyton mentagrophytes, T. ru­brum, T. verrucosum, T. tonsurans, Microsporum canis and Epidermophyton floccosum were finally identified based on the col­ony morphology and microscopic criteria. Specific PCR products and RFLP patterns for Mva I, Hinf I and Hae III en­zymes allowed the rapid identification and reliable differentiation of isolated dermatophytes at the genus or species level for 5-10 day-old colonies. Conclusions: The results showed that PCR-RFLP analysis of the ITS region of rDNA is a rapid and reliable tool which al­lows identification of major pathogenic dermatophytes isolated in this study at species level in young 5-10 day-old colonies.
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نویسندگان مقاله h میرزاحسینی | h mirzahoseini
dept. of biotechnology, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

e امیدی نیا | e omidinia
dept. of biotechnology, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

m شمس قهفرخی | m shams ghahfarokhi
dept. of mycology, faculty of medical sciences, tarbiat modares university, tehran, iran

سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

g صادقی | g sadeghi
dept. of mycology, faculty of medical sciences, tarbiat modares university, tehran, iran

سازمان اصلی تایید شده: دانشگاه تربیت مدرس (Tarbiat modares university)

m رزاقی ابیانه | m razzaghi abyaneh
dept. of mycology, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

نشانی اینترنتی http://ijph.tums.ac.ir/index.php/ijph/article/view/3206
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