Veterinary Research Forum
Vet Res Forum

Development of a diagnostic indirect ELISA test for detection of Brucella antibody using recombinant outer membrane protein 16 kDa (rOMP16)

Document Type : Original Article

Authors

Mehdi Golchin 1
Somayye Mollayi 1
Elham Mohammadi 1
Neda Eskandarzade 2

1 Department of Pathobiology, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

2 Department of Basic Sciences, School of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran

https://doi.org/10.30466/vrf.2021.524590.3141
Abstract
Brucellosis is considered as one of the important global zoonotic diseases that causes medical as well as economic problems especially in tropical countries. The illness has no specific pathognomonic signs; therefore, the rapid and accurate diagnosis of the disease has a very important role in preventing the Brucella spillover and treatment. The purpose of this study was to design a new indirect ELISA test for detection of human brucellosis based on using recombinant Brucella abortus outer membrane protein 16.00 kDa (rOMP16) as an antigen. OMP16 gene of B. abortus was initially synthesized and cloned in pET-21d vector and then expressed in Escherichia coli cells. The expression was confirmed by the SDS-PAGE, western blotting and dot blotting. The purified protein was coated in ELISA plates and an indirect ELISA was performed on 70 human serum samples. The results were evaluated with a commercial IgG ELISA kit and Rose Bengal plate agglutination tests as reference tests. Diagnostic performance of designed OMP16 ELISA test in comparison with Rose Bengal plate test revealed 100% of sensitivity, 95.00% of specificity and good Fleiss kappa agreement, whereas, where it was compared to commercial ELISA kit, it revealed very good kappa agreement with 100% of sensitivity and 100% of specificity in cut-off value of 0.13. It was concluded that OMP 16.00 kDa could be acceptable alternative antigen for detecting Brucella IgG antibody with high accuracy.

Keywords

Brucellosis
Indirect ELISA
OMP16
Sensitivity
Specificity
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Veterinary Research Forum
Volume 13, Issue 3
Summer 2022
Pages 387-391

  • Receive Date 06 February 2021
  • Revise Date 15 March 2021
  • Accept Date 08 May 2021

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