شناسایی مایکوپلاسما بویس در گاوهای مبتلا به ورم پستان بالینی با استفاده از کشت و روش واکنش زنجیره‌ای پلی‌مراز بر اساس ژن‌های 16SrRNA و uvrC

نوع مقاله: علمی پژوهشی

نویسندگان

1 گروه میکروبیولوژی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران

2 آزمایشگاه رفرانس مایکوپلاسما، مؤسسه تحقیقات واکسن و سرم سازی رازی، کرج، ایران.

چکیده

مایکوپلاسما بویس از عوامل اصلی ایجاد پنومونی، ورم پستان و آرتریت مایکوپلاسمائی در گاو می‌باشد. هدف از این مطالعه، شناسایی مایکوپلاسما بویس در گاوهای مبتلا به ورم پستان بالینی با استفاده از کشت و روش زنجیره‌ای پلیمراز بود. این مطالعه روی تعداد 328 نمونه شیر گاو مبتلا به ورم پستان بالینی که در طول یک سال به آزمایشگاه مرجع مایکوپلاسمای مؤسسه تحقیقات واکسن و سرم‌سازی رازی ارجاع می‌شد، اجرا گردید. نمونه‌ها در دمای 4 درجه سلسیوس و در مدت کمتر از 24 ساعت به آزمایشگاه ارسال و به مدت 24-18 ساعت در 37 درجه سلسیوس انکوبه می­شدند و پس از فیلتر شدن به محیط اختصاصی PPLO، در انکوباتور Co2دار به مدت 10-7 روز از نظر رشد پایش می­شدند. همزمانDNA نمونه‌ها به وسیله روش فنل- کلروفرم استخراج و خالص­سازی شده و از روش PCR برای تشخیص جنس مایکوپلاسما بر اساس ژن 16SrRNA و برای تشخیص گونه بویس از جستجوی وجود ژن uvrC استفاده گردید.از تعداد 328 نمونه‌، 58 مورد (69/17 درصد)  در محیط کشت PPLO مثبت شناخته شدند. از نظر حضور ژن مربوط به جنس مایکوپلاسما، تعداد 97 نمونه (57/29 درصد) مثبت شناخته شدند که از این تعداد، 14 نمونه (26/4 درصد) ژن اختصاصی گونه بویس (uvrC) را نشان دادند.نتایج این مطالعه روش PCR را به عنوان یک تست سریع و قابل اعتماد در شناسایی مایکوپلاسما بویس در نمونه‌های شیر معرفی کرده و نشان داد که میزان حضور مایکوپلاسما بویس در گله‌های شیری در ایران بالا است و می‌تواند به عنوان یکی از عوامل اصلی مسبب ورم پستان بالینی در گاو مطرح باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Identification of Mycoplasma bovis in cows with clinical mastitis using culture and Polymerase Chain Reaction based on 16SrRNA and uvrC genes

نویسندگان [English]

  • Mohsen Imandar 1
  • Seyed Ali Pourbakhsh 2
  • Mahmood Jamshidian 1
  • Taghi zahraei salehi 1
1 Department of Microbiology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 Mycoplasma Reference Laboratory, Razi Vaccine and Serum Research Institute, Karaj, Iran.
چکیده [English]

Mycoplasma bovis is one of the main causes of pneumonia, mastitis and arthritis in cattle. The aim of this study was to identify the M. bovis in cows with clinical mastitis using culture and PCR based on 16SrRNA and uvrC genes. A total of 328 milk samples were obtained from cattle with clinical mastitis in industrial dairy herds. Samples were sent to the Mycoplasma reference laboratory, Razi Vaccine and Serum Research Institute at 4°C in less than 24 hr and incubated for 18-24 hr at 37°C. After filtering to specific PPLO medium, they were incubated in the presence of Co2 and were monitored for 7-10 days. Simultaneously, DNA of the samples was extracted and purified by Phenol-Chloroform method and PCR was used to detect mycoplasma genus based on 16SrRNA and detection of M. bovis species based on uvrC genes. Out of 328 samples, 58 samples were positive in PPLO agar media. Ninty-seven samples (29.57%) were positive for mycoplasma gene, of which 14 samples (4.26%) showed specific gene of M. bovis. The results of this study introduced the PCR method as a rapid and reliable test for detection of M. bovis in milk specimens. The results also showed that the presence of M. bovis in dairy herds in Iran is high and it can be considered as one of the main causes of the clinical manifestation of cattle mastitis.

کلیدواژه‌ها [English]

  • Mycoplasma bovis
  • uvrC gene
  • Mastitis
  • Cow
  • PCR
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