Molecular Characterization of canine parvovirus (CPV) antigenic variants from healthy and diarrheic dogs in Urmia region, Iran

Document Type : Infectious agents- Diseases

Authors

1 Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

3 Graduated from the Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

Abstract

Backgrounds: Canine parvovirus (CPV) has been incriminated as a primary pathogen related to acute hemorrhagic enteritis in dogs. Three major antigenic variants of CPV (CPV-2a/2b/2c) have so far been identified. Objectives: This study was carried out to investigate the frequency of CPV-2 and its variants (CPV-2a/2b/2c) in a population of healthy and diarrheic dogs in the north west of Iran. Methods: A total of 35 stool samples from healthy (n=16) and diarrheic (n=19) dogs were screened for all variants (2a, 2b, and 2c) by polymerase chain reaction (PCR) using primer pair 555for/555rev resulting in a PCR product of 583 bp in length. The resulting fragments were further digested by MboII endonuclease that selectively recognizes the restriction site “GAAGA” unique to CPV2c only. All undigested samples were subjected to PCR assays with primer pair Pab (which detects both CPV-2a and CPV-2b types) and primer pair Pb (which detect only CPV-2b type) primer pairs. The relationship of health status, breed, age, sex and vaccination status with PCR results were analyzed using statistical tests. Results: From a total of 35 samples, 10 samples were found to be positive by 555for/555rev primers that were further analyzed by MboII digestion of PCR products. One sample was characterized as CPV-2c and nine samples were categorized as CPV-2a or CPV-2b. All nine undigested samples resulted positive by PCR using Pab primers, out of which 7 resulted positive by PCR using Pb primer pairs, indicating that they are of CPV-2b variant. Conclusions: It seems that CPV-2b is prevalent variant circulating in the North West of Iran. Results also indicated that CPV-2a and CPV-2c are affecting dogs, suggests constant surveillance and monitoring of CPV variants.

Keywords

Article Title [Persian]

شناسایی مولکولی واریانت‌های آنتی‌ژنی پاروویروس سگ (CPV) جدا شده از سگ‌های سالم و اسهالی در منظقه ارومیه، ایران

Authors [Persian]

  • حبیب دستمالچی ساعی 1
  • شهرام جوادی 2
  • شهلا اکبری 3
  • نرجس هادیان 3
  • ادریس زرزا 3
1 گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران
2 گروه علوم بالینی، دانشکده دامپزشکی دانشگاه ارومیه، ارومیه، ایران
3 دانش آموخته دانشکده دامپزشکی دانشگاه ارومیه، ارومیه، ایران
Abstract [Persian]

زمینه مطالعه: پاروویروس سگ ‌(CPV) به عنوان یک جرم بیماری‌زای مهم در ارتباط با انتریت خونریزی دهنده حاد در سگ‌ها قلمداد می‌شود. تاکنون سه واریانت آنتی‌ژنی عمده از CPV (CPV-2a/2b/2c) شناسایی شده است. اهداف: این مطالعه جهت بررسی وفور CPV-2 و واریانت‌های آن (CPV-2a/2b/2c) در جمعیتی از سگ‌های سالم و اسهالی در شمال غرب ایران انجام گرفت. روش کار: در کل 35 نمونه مدفوع از سگ‌های سالم (16=n) و اسهالی (19=n) از نظر حضور همه واریانت-ها (2a، 2b و 2c) بوسیله واکنش زنجیره‌ای پلیمراز (PCR) و با استفاده از جفت پرایمرهای rev555/for555 که محصول PCR 583 جفت بازی تولید می‌نمایند، مورد غربالگری قرار گرفتند. قطعات حاصله با اندونوکلئاز MboII مورد هضم قرار گرفتند که بطور انتخابی محل تعیین حدودی "GAAGA" را شناسایی می‌نماید که تنها منحصر به CPV-2c است. همه نمونه-های هضم نشده تحت آزمایشات PCR با جفت پرایمر Pab (که هر دوی تیپ‌های CPV-2a و CPV-2b را شناسایی می-کند) و جفت پرایمر Pb (که تنها تیپ CPV-2b را شناسایی می‌نماید) قرار گرفتند. ارتباط وضعیت سلامتی، نژاد، سن، جنس و وضعیت واکسیناسیون با نتایج PCR با استفاده از تست‌های آماری آنالیز شدند. نتایج: از کل 35 نمونه، 10 نمونه با پرایمرهای rev555/for555 مثبت یافت شدند که مورد آنالیز بیشتر بوسیله هضم MboII فرآورده‌های PCR قرار گرفتند. یک نمونه به عنوان CPV-2c شناسایی گردید و 9 نمونه به عنوان CPV-2a و CPV-2b طبقه‌بندی شدند. همه 9 نمونه هضم نشده به PCR انجام شده با پرایمرهای Pab نتیجه مثبت دادند، که از این تعداد هفت نمونه به PCR انجام شده با پرایمرهای Pb نتیجه مثبت داده که نشان می‌دهد آنها از واریانت CPV-2b هستند. نتیجه‌گیری نهایی: به نظر می‌رسد که CPV-2b واریانت شایع است که در شمال غرب ایران در حال گردش است. نتایج همچنین نشان دادند که CPV-2a و CPV-2c سگ‌ها را درگیر نموده، که این موضوع نشان دهنده مراقبت مداوم و نظارت واریانت‌های CPV می‌باشد.

Keywords [Persian]

  • واریانتهای آنتیژنی
  • پاروویروس سگ
  • سگ
  • PCR-RFLP
  • تعیین توالی

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Iranian Journal of Veterinary Medicine
Volume 11, Issue 1
January 2017
Pages 9-19

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