Investigation of the seroreactivity of Brucella-infected patients with the recombinant OMP 31 antigen using the ELISA diagnostic method.
Subject Areas : Plasma biomarkers
Taromchi Amir Hossein
1
,
Zahra Taghipour
2
,
ُّFaezeh Sabzehei
3
,
saeid Kaboli
4
1 -
2 - Department of Medical Biotechnology, school of medicine, Zanjan University of Medical Sciences
3 - Department of Medical Biotechnology, school of medicine, Zanjan University of Medical Sciences
4 - Department of Medical Biotechnology, school of medicine, Zanjan University of Medical Sciences
Keywords: Omp31, Brucellosis, Serodiagnosis, ELISA,
Abstract :
Background and Objective: Brucellosis or Malta fever is a zoonotic bacterial infection with a global distribution caused by Brucella species. The hallmark symptoms of this disease include fever in humans and abortion in infected animals. The essential step in controlling this disease is correct and timely identification; therefore, the development of effective diagnostic methods is important. In this study, an ELISA test based on recombinant protein OMP 31 was used, which, based on previous research, is an immunogenic protein with the ability to stimulate an immune response and produce appropriate antibodies. The aim of this project is to investigate the reactivity of this protein with IgG and IgM antibodies present in the serum of patients.
Materials and Methods: The designed gene cassette was cloned in the pET-28a vector and transferred into a bacterial expression host. After confirming the nature of the protein, an appropriate volume of it was expressed and purified by affinity chromatography. After determining the concentration, the antigen was coated on an ELISA plate and its reactivity with patient serum and control serum was examined, and the sensitivity and specificity for both IgM and IgG antibodies were evaluated by ROC analysis.
Results: After expression of the recombinant protein, its identity was confirmed by Western blot and its concentration was estimated at 54.37 μg/ml by Bradford test, and a suitable reaction with the tested sera was reported, with a sensitivity and specificity of 56.56 and 72.5% for IgG; and 75.7% and 62.79 for IgM.
Conclusion: The produced recombinant protein was able to react with patient serum, and a significant difference in its reactivity with the patient group and the control group was observed.
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