نوع مقاله : مقاله پژوهشی
نویسندگان
گروه زیست شناسی، دانشکده علوم پایه، دانشگاه فردوسی مشهد.
چکیده
آلودگیهای باکتریایی مواد غذایی یکی از مهمترین مسائل در زمینه ایمنی مواد غذایی میباشد.لذا استفاده از روشهای تشخیصی سریع و دقیق حائز اهمیت است. یکی از مهم ترین این روشها، تکنیک مولکولی PCR است که امکان تشخیص دقیق و با حساسیت بالا در مدت زمان کم، از مزایای آن است. اما در صورتیکه تشخیص چندین باکتری بیماریزا مد نظر باشد،multiplex PCR (mPCR)بهترین گزینه خواهد بود. در این تحقیق از سه باکتریلیستریا منوسایتوژنز، اشرشیا کلی O157:H7و جنس سالمونلا (سالمونلا انتریکا سرووار تیفی موریوم) که از باکتریهای بیماریزای مهم منتقل شده از طریق آب و مواد غذایی هستند استفاده شد و به کاهو، که نمونه ای از سبزیجات پر مصرف در تهیه سالاد های آماده است، تلقیح و با روش mPCR بطور همزمان شناسایی شدند. بدین منظور مخلوطی از پرایمر های اختصاصی مربوط به باکتریها یعنی، invA( پروتئین تهاجمیA)برای جنس سالمونلا با 284 جفت باز، prfA(پروتئین افزایش دهنده نسخه برداری از ژنهای عامل ویرولانس) برای لیستریا منوسایتوژنزبا 217 جفت باز وrfb (ژن مسئول تولید آنتی ژنO157) برایاشرشیا کلی O157:H7با 420 جفت باز،به همراه DNAاستخراج شده از مخلوط سه باکتری تلقیح شده به کاهو و سایر معرف های واکنشPCR به یک لوله واکنش، منتقل و همزمانPCRشدند. با این روش، امکان تشخیص همزمان سه باکتری بیماریزای فوق با حساسیت cfu/g1امکان پذیر شد.
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