Typing of Toxigenic Isolates of Clostridium perfringens by Multiplex PCR in Ostrich
محورهای موضوعی : CamelE. Zandi 1 , M.R. Mohammadabadi 2 , M. Ezzatkhah 3 , A.K. Esmailizadeh 4
1 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
2 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
3 - Razi Vaccine and Serum Research Institute, Kerman, Iran
4 - Department of Animal Science, Shahid Bahonar University of Kerman, Kerman, Iran
کلید واژه: Multiplex PCR, biochemical tests, genetic typing, <i>Clostridium perfringens</i>,
چکیده مقاله :
Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five various types, each of which capable of causing a distinct disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also show low sensitivity and specificity. The aim of this research was to identify the unlike types of Clostridium perfringens using PCR method. In this study, 120 ostrich-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. Clostridium perfringens isolated types were divided as follows: 100% type A, 0% type B, 0% type C and 0% type D. It should be emphasized that, up to now, Clostridium perfringens type A has not been reported in Iran.
کلستریدیوم پرفرینجنس یک پاتوژن مهم است که بیماریهای گوناگون زیادی را ایجاد میکند. این باکتری دارای 5 تیپ مختلف است که هر تیپ قادر به ایجاد بیماری مشخصی میباشد. روشهای مختلفی برای شناسایی باکتریها وجود دارند که بیشتر آنها مشکل، زمان بر، پرهزینه و دارای حساسیت و اختصاصیت پایینی هستند. هدف این پژوهش تعیین انواع نامشابه کلستریدیوم پرفرینجنس با روش PCR بود. در این مطالعه، نمونه‌های مدفوع 120 شترمرغ به صورت تصادفی از نواحی اطراف شهر کرمان واقع در جنوب شرقی ایران جمع‌آوری شدند. بعد از عمل‌آوری و کشت نمونه‌ها، کلونی‌های تولید شده از نظر مورفولوژیکی مطالعه شدند، تست رنگ آمیزی گرم نیز انجام شد و جنس این باکتریها با استفاده از تست‌های بیوشیمیایی تعیین گردید. DNA استخراج شده از باکتریهای جدا شده برای تعیین ژنوتیپ توسط multiplex PCR با آغازگرهای اختصاصی آزمون شد. بر اساس طول قطعات سنتز شده توسط PCR، تیپ‌های توکسین و استرین‌های باکتریایی شناسایی شدند. تیپ‌های جدا شده کلستریدیوم پرفرینجنس عبارت بودند از: 100 درصد تیپ A، 0 درصد تیپ B، 0 درصد تیپ C و 0 درصد تیپ D. این نکته را می‌توان مورد تأکید قرار داد که تا کنون تیپ A کلستریدیوم پرفرینجنس گزارش نشده است.
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