این سایت در حال حاضر پشتیبانی نمی شود و امکان دارد داده های نشریات بروز نباشند
Middle East Journal of Digestive Diseases، جلد ۱۱، شماره ۳، صفحات ۰-۰
عنوان فارسی
چکیده فارسی مقاله
کلیدواژه‌های فارسی مقاله
عنوان انگلیسی A Two-Step Approach for Diagnosing Glutamate Dehydrogenase Genes by Conventional Polymerase Chain Reaction from Clostridium difficile Isolates
چکیده انگلیسی مقاله BACKGROUND: Clostridium difficile is the major causative agent of nosocomial antibiotic-associated colitis. The gold standard for C. difficile detection is stool culture followed by cytotoxic assay, although it is laborious and time-consuming. We developed a screening test based on a two-step conventional polymerase chain reaction (PCR) approach to detect glu D, the glutamate dehydrogenase ( GDH) enzyme gene, which is a marker for screening of C. difficile . Targeting glu D comparing to the conserved stable genetic element of pathogenicity locus (PaLoc), with an accessory gene of Cdd 3, was an effective method for the detection of this pathogen from patients with enterocolitis. METHODS: Fresh fecal samples of the patients who were clinically suspicious for antibiotic-associated colitis were collected. Stool specimens were cultured on the cycloserine-cefoxitin fructose agar (CCFA) in an anaerobic condition, following alcohol shock treatment and enrichment in Clostridium difficile Brucella broth (CDBB). On confirmed colonies, PCR was carried out for detection of PaLoc subsidiary gene, Cdd 3, and toxicogenic genes, tcd A and tcd B. The glu D that is GDH gene detection was performed by conventional PCR on the extracted DNA from 578 fresh stool samples. RESULTS: 57 (9.8%) strains of C. difficile were approved by conventional PCR for glu D and Cdd 3 genes, in which 37 (6.4%) colonies had tcd A+/ tcd B+ genotype, 2 (0.3%) tcd A+/ tcd B-, 4 (0.7%) tcd A-/ tcd B+ and the remaining 14 (2.4%) colonies were tcd A and tcd B negative. CONCLUSIONS: These results demonstrate that targeting glu D by PCR is quite promising for rapid detection of C. difficile from fresh fecal samples. Furthermore, the multiple-gene analysis for tcd A and tcd B assay proved a reliable approach for diagnosing of toxigenic strains among clinical samples.
کلیدواژه‌های انگلیسی مقاله
نویسندگان مقاله | Sepideh Khodaparast
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University. Tehran, Iran


| Ashraf Mohabati Mobarez
Digestive Disease Research Institute, Tehran University of Medical Sciences, Tehran, Iran


| Mehdi Saberifiroozi


نشانی اینترنتی http://mejdd.org/index.php/mejdd/article/view/1982
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/332/article-332-1674710.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده
نوع مقاله منتشر شده Original Article
برگشت به: صفحه اول پایگاه   |   نسخه مرتبط   |   نشریه مرتبط   |   فهرست نشریات