تعیین تیپهای ایزولههای Clostridium perfringens به روش Multiplex PCR در شتر مرغ

Subject Areas : Camel

E. 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

Received: 2014-01-11 Accepted : 2014-04-15 Published : 2014-12-01

Keywords: Multiplex PCR, کلستریدیوم پرفرینجنس, تست‌های بیوشیمیایی, تعیین تیپ ژنتیکی,

Abstract :

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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