تشخیص مولکولی و میکروسکوپی گونه‌های آناپلاسما در شترهای کشتار شده در استان گلستان

نوع مقاله : مقاله کامل

نویسنده

عضو هیات علمی موسسه تحقیقات واکسن و سرم‌سازی رازی، سازمان تحقیقات و آموزش کشاورزی، کرج، ایران

چکیده

آناپلاسما میکروارگانیسم درون سلولی اجباری است که در سلول‌های خونی میزبان زندگی می‌کند و منجر به بیماری و تحمیل هزینه‌های مراقبتی زیادی می‌شود. هدف از مطالعه حاضر بررسی مولکولی و میکروسکوپی حضور جنس و گونه‌های آناپلاسما در شترهای استان گلستان بود. نمونه‌های خون از شترهای کشتار شده در کشتارگاه صنعتی اینچه برون برای مدت دو سال جمع‌آوری شد. بررسی میکروسکوپی با تهیه گسترش‌های خونی رنگ‌آمیزی شده با گیمسا صورت گرفت. آزمون مولکولی با تکثیر اولیه قطعات DNA و تأیید آن‌ها با انجام nested-PCR صورت گرفت. از پرایمرهای اختصاصی بر روی ژن‌های 16S rRNA و msp4 برای تکثیر گونه‌های مختلف آناپلاسما استفاده شد. قطعات تکثیری حاصل از PCR اولیه و nested-PCR حضور جنس آناپلاسما در 33 نمونه از 90 نمونه (67/36 درصد) را مثبت نشان داد. در میان نمونه‌های حاوی جنس آناپلاسما، 22/2 درصد، 22/22 درصد و 33/13 درصد با PCR اختصاصی به‌ترتیب گونه‌های آناپلاسما مارجیناله، آناپلاسما فاگوسیتوفیلم و آناپلاسما بویس شناسایی شدند. هیچ کدام از نمونه‌ها با PCR اختصاصی حضور گونه آناپلاسما سنتراله را نشان ندادند. مشاهده میکروسکوپی اجرام آناپلاسمایی درون گلبول‌های قرمز حاکی از حضور جنس آناپلاسما در 33/33 درصد از گسترش‌های خونی بود. با مقایسه آزمون مولکولی و میکروسکوپی حساسیت، اختصاصیت و صحت برای شناسایی جنس آناپلاسما به‌ترتیب 21/21 درصد، 67/66 درصد و 50 درصد بدست آمد. نتایج مطالعه حاکی از حضور آناپلاسما در میان شترهای کشتار شده در استان گلستان بود. بر طبق نتایج، بررسی مولکولی آناپلاسما نتایج دقیق‌تری نسبت به بررسی میکروسکوپی بدست می‌دهد.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular and microscopy detection of Anaplasma species in camel of Golestan province

نویسنده [English]

  • parastoo poorghafoor langroodi
Member of Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
چکیده [English]

Anaplasma is an intracellular microorganism that lives in blood cells of host, causing illness and impose care costs. This study was aimed to investigate molecular and microscopic presence of genus and Anaplasma species in camels of Golestan province. Blood samples were collected from adult slaughtered camels in Inche Bourun industrial slaughterhouse for two years. Microscopic examination was performed by preparing Giemsa-stained blood smear. Molecular test was performed by primary amplification of DNA fragments and then they confirmed by performing the nested-PCR. The specific primers of 16S rRNA and msp4 genes were used to detect different species of Anaplasma. The proliferation of the primary and the nested PCR products showed that genus Anaplasma in 33 out of 90 samples (36.37 percent) were positive. Among samples including Anaplasma 2.22, 22,22 and 13.33 percentage were detected by specific PCR for A. marginale, A. phagocytophilum and A. bovis, respectively. None of the samples showed the presence of Anaplasma centrale. Microscopic observation of anaplasmic bodies in red blood cells (RBC) indicated the presence of Anaplasma genus in 33.33 percent of blood smears. By comparing molecular and microscopic tests, the sensitivity, specificity and accuracy to identify the genus Anaplasma were 21.21, 66.67 and 50 percent, respectively. The results of this study showed that Anaplasma was present among camels in Golestan province. According to the results, molecular study of Anaplasma yields more accurate results than microscopic examination.

کلیدواژه‌ها [English]

  • Anaplama
  • 16S rRNA gene
  • msp4 gene
  • Blood smear
  • Golestan
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