عیارسنجی آنتی توکسین اپسیلون کلستریدیوم پرفرینجنز تیپ D در سرم خرگوش به روش الایزای غیر مستقیم

نوع مقاله : مقاله کامل

نویسندگان

بخش تحقیق و تولید واکسن های باکتریایی بی هوازی، آزمایشگاه تحقیقاتی کلستریدیا، موسسه تحقیقات واکسن و سرم‌سازی رازی، سازمان ترویج آموزش توسعه کشاورزی، تهران، ایران

چکیده

کلستریدیوم پرفرینجنز یک باکتری گرم مثبت که براساس چهار توکسین اصلی آلفا، بتا، اپسیلون و یوتا به پنج تیپ A تا E تقسیم می شود. توکسین اپسیلون از توکسین‌های قوی کلستریدیایی و توکسین اصلی تیپ‌های B و D و عامل بیماری گاز گانگرن درانسان و آنتروتوکسمی در دام است. ارزیابی ایمنی ناشی از واکسن که در پیش‌گیری از این بیماری از اهمیت خاصی برخودار بوده که در فارماکوپه روش
(SN (Serum neutralization test برای آن پیشنهاد گردیده است. هدف این پژوهش ارزیابی تست الایزا به عنوان روشی جایگزین برای اندازگیری آنتی‌توکسین اپسیلون می‌باشد. ابتدا تست اندازه‌گیری توانایی واکسن به روش معمول انجام یافته و سرم‌های اخذ شده با هر دو روش معمول و الایزای غیرمستقیم بررسی گردید. میزان cut off کنترل منفی در تست الایزا 42/۰ برآورد شد و نتایج آزمون کراس نشان داد که سیستم الایزا برای توکسین اپسیلون اختصاصی عمل می‌نماید نتایج این پژوهش ارتباط معنی‌داری بین این دو تست در موارد واکسینه شده با نمونه فرمانتور نشان داد. آنالیز رگرسیون خطی، همبستگی ۸۴/۰ با P≤0.01 را نشان داد. در نهایت پیشنهاد گردید از تست الایزا به عنوان جایگزینی برای ارزیابی سطح ایمنی حیوانات آزمایشگاهی استفاده نمود. اگرچه برای استفاده در حیوانات هدف نیاز به تحقیقات بیشتر در این زمینه ضروری می باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation of Epsilon Antitoxin of Clostridium Perfringens Type D in The Rabbit Serum By Indirect ELISA

نویسندگان [English]

  • A. R. Paradise
  • L. Abdolmohammadi Khiav
Department Anaerobic Bacterial Vaccine Production and Research, Clostridia Research Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran.
چکیده [English]

Clostridium perfringens is a gram-positive bacterium divided into five major A, E, and iota toxins based on the four main toxins. Epsilon toxin is a potent clostridial toxin and a major toxin of Clostridium Perfringens B and D types, and is the cause of gangrene gas in humans and enterotoxemia in human. For safety assessment of the vaccine, which is important for prevention of this disease, has been proposed SN (Serum neutralization test) method in the pharmacopoeia. The purpose of this study was to evaluate the ELISA as an alternative method for measuring of epsilon antitoxin. First, vaccine potency test was performed using conventional method and the sera obtained were measured using both conventional and ELISA methods. The negative control cut-off was calculated 0.42 and cross-examination was shown that epsilon toxin had no cross reaction with other toxins of Clostridium spp.
The results of this study showed that there is a significant agreement between two tests for serum samples of vaccinated rabbits by polyvalent vaccine. Linear regression analysis gave correlation coefficients of 0.697 for the indirect ELISA, with a significance level of P<0.01. Finally, ELISA test could be used as an alternative MNT test which used for detection antibody in laboratory animals. However, further research in this field is needed for target animals.

کلیدواژه‌ها [English]

  • Indirect ELISA
  • enterotoxaemia vaccine
  • Clostridium perfringens type D
  • epsilon antitoxin
  • serum neutralization test
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