جداسازی، شناسایی و آنالیز فیلوژنی برمبنای ژن F ویروس بیماری نیوکاسل جدا شده از مرغداری های گوشتی استان مرکزی

نوع مقاله : مقاله کامل

نویسندگان

1 موسسه تحقیقات واکسن و سرم سازی رازی شعبه اراک، سازمان تحقیقات، آموزش و ترویج کشاورزی، اراک، ایران

2 موسسه تحقیقات واکسن و سرم سازی رازی ، سازمان تحقیقات، آموزش و ترویج کشاورزی،کرج، ایران

3 موسسه تحقیقات واکسن و سرم سازی رازی ، سازمان تحقیقات، اموزش و ترویج کشاورزی،کرج، ایران

چکیده

بیماری نیوکاسل از مسری‌ترین بیماری‌های ویروسی ماکیان با انتشار سریع در سراسر جهان است که با علایم تنفسی، اختلالات عصبی و گوارشی، و تلفات زیاد مشخص می‌شود. ابتلا به این بیماری تاثیر منفی در تجارت صنعت طیور، فرآورده‌های آن‌ها در کشورهای درحال توسعه و کشورهای توسعه یافته می‌گذارد. عامل جزء خانواده پارامیکسوویریده، جنس Avulavirus بوده ویروس های مسبب نیوکاسل متعلق به avian paramyxovirus type 1 (APMV-1)  می‌باشند. هدف از این مطالعه شناسایی، آنالیز فیلوژنی جدایه‌های ویروس نیوکاسل در مرغداری‌های استان مرکزی با استفاده از روش RT- PCR بود. در این تحقیق که با هدف شناسایی و آنالیز فیلوژنی بر اساس ژن F صورت پذیرفت، 30 نمونه شامل مغز،نای، ریه و طحال از مرغ های مبتلا در مزارع مشکوک به بیماری اخذ گردید. نمونه ها پس از آماده سازی در تخم مرغ SPF  جنین دار 9 تا 11 روزه تلقیح گردیدند.پس از 2 تا 5 روز مایع آلانتوئیک تخم‌مرغ‌های مورد آزمایش برداشت شد وجود ویروس به روش HA مورد ارزیابی قرار گرفت. مرحله بعد نمونه‌ها با استفاده از پرایمر اختصاصی ژن F (1350bp) تحت عملیات RT-PCR قرار گرفت. در این مطالعه تعداد 13 جدایه نیوکاسل مورد تایید قرارگرفت. آنالیز فیلوژنتیکی و رسم درخت  فیلوژنی با استفاده از نرم‌افزار MEGA صورت پذیرفت. نتایج نشان داد کلیه جدایه‌ها متعلق به ژنوتیپ VII و زیرژنوتیپ VII j می‌باشند. داده‌های بدست آمده جهت گسترش کارآیی برنامه واکسیناسیون و پیشگیری از بیماری و همچنین بکارگیری آن در مسیر ساخت واکسن برعلیه ویروس عامل بیماری نیوکاسل بنیادی می‌باشد. 

کلیدواژه‌ها

عنوان مقاله [English]

Identification and phylogenetic analysis based on Newcastle disease virus gene F isolated from broiler poultry farms in Markazi province

نویسندگان [English]

  • M. Ahmadi 1
  • M. M. Ebrahimi 2
  • Sh. Shahsavandi 3
  • S. Ebrahimi 1
  • A. Hamidi 1
  • Sh. Ghaemmaghami 1
1 Agricultural Research Education and Extension Organization, Razi Vaccine and Serum Research Institute, Arak Branch , Iran
2 Agricultural Research Education and Extension Organization, Razi Vaccine and Serum Research Institute, Karaj, Iran
3 Agricultural Research Education and Extension Organization, Razi Vaccine and Serum Research Institute, Karaj, Iran
چکیده [English]

Newcastle Disease Virus (NDV)is one of the most contagious viral diseases of poultry with rapid spread around the world. It identified by respiratory symptoms, nervous and digestive disorders, and high mortality. NDv causes significant economic losses in poultry industry throughout the world. Avian paramixovirus serotype 1(APMV-1) is a member of family paramyxoviridae, the genus Avulavirus, Newcastle disease virus belongs to APMV-1. The aim of this study to recognize and phylogenic analysis of Newcastle disease Virus isolated from poultry farms in Markazi Province by RT-PCR. In this research, recognition and phylogenic analysis was performed based on F gene of Newcastle disease virus. About 30 samples including brain, trachea, lung and spleen were obtained from suspected farms. Then, They were inoculated to 9 - 11 days age SPF embryonic eggs. After that 2-5 days, The Allantoic fluid harvested, the HA test was performed and genetically analysis using specific primer for F gene was done by RT PCR. In this study, Among the samples 13 of them was confirmed as Newcastle disease virus. Phylogeny tree was constructed for F gene using MEGA 6 software. The results showed that all isolates NDV belonged to genotype VII and subgenotype VII-j. These obtained data can use to improve the efficacy of the vaccine and disease prevention program, as well as its use for vaccine production against the Newcastle-disease causative agents.

کلیدواژه‌ها [English]

  • Newcastle disease
  • HA
  • phylogeny
  • isolation
  • inoculation
  • RT-PCR

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سابقه مقاله

  • تاریخ دریافت: 21 دی 1397
  • تاریخ بازنگری: 17 بهمن 1398
  • تاریخ پذیرش: 30 فروردین 1399

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