طراحی یک روش الایزای رقابتی جهت ارزیابی قدرت سرم ضد سم مار

نوع مقاله: مقاله کامل

نویسندگان

1 موسسه تحقیقات واکسن وسرم سازی رازی بخش پروتئومیکس وبیوشیمی

2 بخش پروتئومیکس و بیوشیمی، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، ترویج و آموزش کشاورزی، کرج، ایران.

3 گروه میکروبیولوژی، دانشکده دامپزشکی، دانشگاه آزاد اسلامی واحد علوم و تحقیقات تهران، تهران، ایران.

4 موسسه تحقیقات واکسن وسرم سازی رازی بخش تصفیه سرم

5 موسسه تحقیقات واکسن وسرم سازی رازی بخش جانوران سمی

چکیده

جهت تعیین توان پتانسی سرم پلی والان ضد سم مار در خنثی کردن سم، از مجموع سموم مارهای ناجا ناجا اکسیانا، ویپرا لبتینا، ویپرا آلبیکورنوتا، اکیز کاریناتوز، پسیدوسراستوس پرسیوس و آجیسترودون هالی استفاده شده است. LD50 یا دوز کشنده در موش µg 5/6 در هر موش تعیین شده است. روش خنثی‌سازی سم جهت تعیین پتانسی سرم ضدسم مار، الایزای رقابتی و LD50 (یک روش درون تن) است. از این دو روش جهت تعیین پتانسی در 15 نمونه سرم اسب هایپرایمن استفاده شد. نتایج الایزای رقابتی برای تعیین پتانسی نشان داد که رقت 12000/1 سرم اسب هایپرایمن توان مهار 50 درصد سم را دارد. الایزای رقابتی با سنجش بیولوژیکی رایج LD50 که یک روش درون تن است مورد مقایسه قرار گرفت. همبستگی معناداری بین تیترهای الایزا و مقادیرLD50 در سطح معناداری 01/0≥P و 95/0=r مشاهده گردید، که نشان می‌دهد روش مورد مطالعه در این پژوهش در شرایط in vitro (برون تن) می‌تواند ظرفیت خنثی سازی آنتی بادی‌های سرم را همانند سنجش بیولوژیک in vivo (درون تن) با دقت بالا تخمین بزند. نتایج پژوهش حاضر نشان می‌دهد روش الایزای رقابتی با قابلیت اندازه گیری مقدار کمپلکس آنتی ژن- آنتی بادی می‌تواند جایگزین مناسبی برای روش خنثی‌سازی کشندگی در محیط درون تن باشد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Designing a competitive ELISA for evaluation of anti-snake venom serum potency

نویسندگان [English]

  • F. Golchinfar, 1
  • R. Madani, 2 3
  • T. Emami, 1
  • H. Zoulfagharian, 4
  • A. Zare, 5
  • N. Mouhammadpour, 5
1 Dept. of Biochem & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
2 Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
3 Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. | Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.
4 Dept. of Serum & Proteomics, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
5 Dept. of Venomous Animals and Antivenom Production, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
چکیده [English]

The venom of Naja naja oxiana, Vipera lebetina, Vipera albicornouta, Echis carinatus, Pseadocerastus persicus and Agkistrodon halys snakes are used to determine the potency of the polyvalent sera in neutralizing the venom. LD50 value per mouse was determined 6.5 μg. Neutralizations methods for obtaining potency of anti venom are competitive ELISA (Enzyme linked immunosorbent assay) and in vivo assay (ED50). Both of these tests were performed to estimate the serum potency in 15 samples of hyper-immune equines. Results of competitive ELISA showed that 1/12000 diluted serum can inhibits 50 percent of antigens. Competitive ELISA was compared with current biological assay ED50. Significant correlations between ELISA titers and values of ED500 at level of P≤ 0.01 and r=0.95 was observed, that indicates competitive ELISA can estimate antibody neutralizing capacity of the serum as well as the in vivo assay. The results of present study shows C-ELISA that measures Ag-Ab complex can used as a suitable replacement method for lethal neutralizing in vivo method.

کلیدواژه‌ها [English]

  • ELISA
  • Snake venom
  • Potency
  • Neutralization
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