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:: Volume 7, Issue 2 (Vol.7, No.2,Summer 2019) ::
J Neyshabur Univ Med Sci 2019, 7(2): 15-22 Back to browse issues page
Design and optimization of Dot-blot ELISA system using specific antigens for simple and rapid diagnosis of glanders
Sajjad Yazdansetad Mr.1 , Nader Mosavari * 2, Keyvan Tadayon Dr.3 , Iraj Mehregan1
1- Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2- Department of Tuberculin & Mallein, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran , nmosavari@gmail.com
3- Department of Veterinary Aerobic Bacteria Vaccines, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
Abstract:   (414 Views)
Introduction
Glanders is one of the oldest contagious and dangerous zoonotic diseases manifesting ulcerative granulomatous lesions on the skin and mucous membranes. Early methods possessing desirable sensitivity and specificity is important to diagnose the disease considering the just only one case report and preventing disease by identification and eradication. The present study was aimed to design and optimize Dot-blot ELISA system using specific antigens for simple and rapid detection of glanders using equine sera samples.
Materials & Methods
Burkholderia mallei strains were cultured in nutrient broth supplemented with glycerol 4%. Whole cell antigens of bacterial strains were precipitated by trichloroacetic acid (TCA) followed by sonication method. The optimum concentrations of antigen-antibody were determined by checkerboard titration. The nitrocellulose membrane was coated by antigen, then stopped by skimmed milk as a blocker. The suspicious equine serum was added to the membrane following HRP-conjugated antibody, the signals were detected by TMB as a substrate.
Results
4 out of 90 sera samples were positive by Dot-blot ELISA. The specificity and sensitivity of the test were evaluated 96.62% and 100%, respectively. B. mallei culture and isolation from clinical specimens of the horses validated the test.
Conclusion
Our study showed that the Dot-blot ELISA is specific and sensitive in glanders diagnosis and it seems to be practical and efficient test due to the rapid, easy interpretation without any special tools, cost-effective and also field-friendly.
Keywords: Burkholderia mallei, glanders, Dot-blot ELISA, antibody
Full-Text [PDF 1232 kb]   (73 Downloads)    
Type of Study: Short Communication | Subject: Veterinary
Received: 2019/05/14 | Revised: 2019/07/6 | Accepted: 2019/06/10 | ePublished ahead of print: 2019/06/11 | Published: 2019/07/6 | ePublished: 2019/07/6
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Yazdansetad S, Mosavari N, Tadayon K, Mehregan I. Design and optimization of Dot-blot ELISA system using specific antigens for simple and rapid diagnosis of glanders. J Neyshabur Univ Med Sci. 2019; 7 (2) :15-22
URL: http://journal.nums.ac.ir/article-1-729-en.html


Volume 7, Issue 2 (Vol.7, No.2,Summer 2019) Back to browse issues page
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