مطالعه فیلوژنی جدایه های Theileria annulata از استان البرز با استفاده از توالی ژن های 18S rRNA و ITS1-2

افشاری, اصغر; حبیبی, غلامرضا; اسماعیل‌نیا, کسری; شایان, پرویز

doi:10.22092/vj.2015.100949

مطالعه فیلوژنی جدایه های Theileria annulata از استان البرز با استفاده از توالی ژن های 18S rRNA و ITS1-2

نوع مقاله : مقاله کامل

نویسندگان

¹ کارشناس ارشد موسسه تحقیقات واکسن وسرم سازی رازی

² استادیار موسسه تحقیقات واکسن وسرم سازی رازی

³ استاد دانشکده دامپزشکی دانشگاه تهران

10.22092/vj.2015.100949

چکیده

تیلریوز گرمسیری بیماری انگلی ناشی از تک یاخته T.annulata بوده و بوسیله کنه های جنس هیالوما منتقل می شود. تشخیص بیماری معمولاً با مشاهده انگل در گسترش های خون محیطی، طحال و غدد لنفاوی صورت می گیرد. امروزه روش های ملکولی با حساسیت و ویژگی بالائی به منظور تشخیص و شناسائی عامل بیماری در دام های مبتلا و ناقل، مطالعات همه گیری شناسی و بررسی های فیلوژنی مورد استفاده قرار می گیرد. با توجه به حضور تیلریوز گاوی در کشور، مطالعه فیلوژنی T.annulata با استفاده از بررسی توالی ژن 18S rRNA و نواحی ITS شامل (5.8S rRNA , ITS2, ITS1) مربوط به دو جدایه استان البرز به همراه رده سلولی سویه واکسن تیلریوز گاوی مورد توجه قرار گرفت. توالی های مورد نظر ابتدا با PCR تکثیر شده و تعیین توالی گردید. درخت فیلوژنی بر اساس توالی های ناحیه ITS و ژن 18S rRNA ترسیم شد. نتایج این مطالعه نشان داد که ژن 18S rRNA دارای ارزش شناسائی در حد جنس و توالی ITS در حد گونه می باشد. یافته های حاصل از بررسی درخت فیلوژنی توالی کامل ژن 18S rRNA نشان داد گونه های بیماری زای T.lestoquardi، T.annulata و T.parva در یک خوشه و مستقل از سایر گونه های تیلریا قرار می گیرند.

20.1001.1.24235407.1393.27.4.13.2

عنوان مقاله [English]

Phylogenetic study of Theileria annulata isolates from Alborz province by using 18S rRNA gene and ITS regions sequences

نویسندگان [English]

A. Afshari ¹
Gh.R. Habibi ²
K. Esmaeilnia ²
P. Shayan ³

¹ Msc Razi Vaccine and Serum Research

² Assistant Professor Razi Vaccine and Serum Research

³ Professor College of Veterinary Tehran University

چکیده [English]

Tropical theileriosis is a tick-transmitted disease which caused by obligative intracellular protozoan Theileria annulata. Diagnosis is usually based on identification of parasite in Giemsa stained smears of peripheral blood, spleen or lymph nodes. Today, the molecular methods with high sensitivity and specificity are used for diagnosis of infected and carrier animals, epidemiological and phylogenetic studies. Due to the presence of bovine theileriosis in Iran, T. annulata phylogenetic analysis has been focused using 18S rRNA gene and ITS region sequences (ITS1, 5.8S rRNA, ITS2) for two Alborz Theileria annulata isolates and bovine theileriosis vaccine strain S15 cell line. The target sequences were amplified by PCR and further PCR product sequencing. The phylogenetic tree were constructed based on 18S rRNA complete gene sequences and ITS region, then the sequences of Iranian isolates and registered Theileria sequences from other countries registered in GenBank were compared. The obtained results showed that the 18S rRNA gene and ITS sequence are valuable for detection and classification at the species and subspecies levels, respectively. Other important finding obtained from the present study was the result from phylogenetic tree of complete 18S rRNA gene sequences that revealed the highly pathogenic Theileria spp. (lymphoproliferative Theileria) including T. annulata, T. lestoquardi and T. parva were placed in a separate cluster away from the other Theileria spp.

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