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Restriction Fragment Length Polymorphism Typing of Staphylococcus aureus Strains Isolated from Bovine Mastitis and Dairy Products in Ahvaz, Iran, Using of Digested Coagulase Gene | ||
| Archives of Razi Institute | ||
| مقاله 11، دوره 74، شماره 3، آذر 2019، صفحه 303-311 اصل مقاله (206.66 K) | ||
| نوع مقاله: Short Communication | ||
| شناسه دیجیتال (DOI): 10.22092/ari.2017.115900.1159 | ||
| نویسندگان | ||
| D. Gharibi1؛ R. Ghadimipour* 1، 2؛ M. Ghorbanpoor1؛ A. Fazlara3 | ||
| 1Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| 2Department of Research and Development, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization , Ahvaz, Iran | ||
| 3Department of Food Hygiene, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran | ||
| چکیده | ||
| Staphylococcus aureus is a major pathogen in the transmission of diseases from animals to humans and vice-versa.Various infections, such as mastitis in cattle, sheep and goats, as well as gastroenteritis due to food poisoning in humans are the most frequent problems caused by S. aureus. The bacteria also lead to severe economic losses in dairy industry. A major virulence factor for the organism is encoded by the coagulase (coa) gene. This study aimed to assess the polymorphisms of the coa gene in S. aureus strains isolated from bovine mastitis and dairy product samples in Ahvaz, Iran. The results showed that out of 91 S. aureus, 80 (87.91%) isolates were positive for coa gene(s). In total, nine different polymerase chain reaction (PCR) products were obtained for coa-positive isolates. A single band was detected in coa PCR with a size ranges from 370 to 830 bp in most isolates (n=77, 96.25%). For three isolates (3.75%), two amplification products were obtained. A PCR product of an estimated size of 590 bp was most frequent, as obtained for 48 (60.00%) isolates. Whereas, 370 and 830 bp PCR products were the least presented, for two (2.50%) and one (1.25%) isolate, respectively. Subsequently, for restriction fragment length polymorphism (RFLP), typing of coa gene and AluI restriction enzyme were used for the digestion of the products. AluI for most of PCR products generated a unique pattern; however, four PCR products (the sizes ranged 750, 670, 590, and 510 bp) generated three or more patterns. Based on AluI RFLP of coa gene, the isolates were classified into 23 groups. Two groups of isolates were dominant, making 45% of the total. According to the findings, one or two types of coa RFLP were dominant among samples that were infected with more S. aureus isolates belonging to different coa RFLP types. | ||
| کلیدواژهها | ||
| Staphylococcus aureus؛ Coagulase Gene؛ Polymerase Chain Reaction؛ Restriction Fragment Length Polymorphism؛ Bovine Mastitis؛ Dairy Products؛ Iran | ||
| عنوان مقاله [English] | ||
| تایپینگ سویههای استافیلوکوکوس اورئوس جداشده از موارد ورم پستان گاوی و محصولات لبنی شهر اهواز با بررسی پلیموفیسم طول قطعات حاصل از هضم ژن کواگولاز | ||
| چکیده [English] | ||
| استافیلوکوکوس اورئوس یکی از عوامل بیماریزای شناختهشده حیوانات و انسان به شمار میرود. عفونتهای مختلفی مانند ورم پستان در گاو، گوسفند و بز، و نیز مسمومیت غذایی و گاستروانتریت در انسان از مشکلات شایعی هستند که توسط این باکتری ایجاد میشوند. استافیلوکوکوس اورئوس همچنین منجر به خسارتهای شدید اقتصادی در صنایع لبنی میشود. ژن کواگولاز (coa) یکی از مهمترین فاکتورهای حدت این ارگانیسم محسوب میشود. مطالعه حاضر با هدف بررسی پلیمورفیسم این ژن در جدایههای استافیلوکوکوس اورئوس حاصل از موارد ورم پستان گاوی و محصولات لبنی شهر اهواز در جنوب غرب ایران طراحی گردید. آزمایش مولکولی 91 جدایه تحت مطالعه نشان داد که 80 جدایه (91/87%) واجد ژن کواگولاز میباشند. در مجموع نه الگوی واکنش زنجیرهای پلیمراز از تکثیر ژن کواگولاز این جدایهها به دست آمد که الگوی اکثر جدایهها(n=77, 96.25%) شامل یک باند 830-370 جفت بازی (bp) بود، در حالی که تعداد کمی از آنها (n=3, 3.75%) محصول واکنش زنجیرهای پلیمراز دو باندی را نشان دادند. یافتههای ما همچنین مشخص نمود که 48 جدایه (00/60%) باندی با اندازه تقریبی 590 جفت باز را تولید کردند، ولی محصولات 370 و 830 جفت بازی به ترتیب فقط برای دو (50/2%) و یک جدایه (25/1%) تشخیص داده شدند. در ادامه بررسیها از آنزیم محدودالاثر AluI برای هضم محصولات واکنش زنجیرهای پلیمراز تکثیر ژن کواگولاز استفاده گردید. بر اساس این آزمایشها برخی از محصولات واکنش زنجیرهای پلیمراز فقط یک پروفایل تولید کردند، در صورتی که چهار محصول 750، 670، 590 و 510 جفت بازی منجر به ایجاد سه و یا تعداد بیشتری پروفایل گردیدند. بطور کلی با ارزیابی پلیمورفیسم طول قطعات حاصل از هضم (RFLP)، تنوع بالایی در ژن مورد مطالعه مشاهده شده و جدایهها در 23 پروفایل مختلف طبقهبندی گردیدند، با این همه در مجموع 45% آنها متعلق به دو پروفایل بودند. بر اساس یافتههای این مطالعه اگر چه نمونههای مورد آزمایش آلوده به سویههایی از استافیلوکوکوس اورئوس بودند که از نظر پلیمورفیسم ژن کواگولاز به ژنوتیپهای متعددی تعلق داشتند ولی اغلب این جدایهها به یک یا دو ژنوتیپ غالب مربوط میشدند. | ||
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