Isolation and purification of the Burkholderia mallei antigenic proteins and its use in diagnostic tests

Document Type : Original Articles

Authors

1 Department of Biology, Payame Noor University, Tehran, Iran

2 Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization (AREEO), Tehran, Iran

10.22092/ari.2024.367086.3346

Abstract

Glanders is a contagious infectious disease caused by Burkholderia mallei that affects both solipeds and carnivores. This disease occasionally leads to human infection through direct contact between humans and infected animals. The recent rise in the prevalence of glanders has caused a focus on control and eradication programs, emphasizing accurate diagnosis of infected cases using a high-performance test. To this end, antigenic proteins were purified from Burkholderia mallei, and the mallein test was then optimized using the purified proteins. Finally, the efficacy of antigenic proteins was evaluated using the complement fixation test (CFT). The laboratory strain of B. mallei was selected and the proteins of inactivated bacteria were precipitated using ammonium sulfate (AS) and trichloroacetic acid (TCA). An optimal precipitation method was selected, and the proteins were purified using size exclusion chromatography (SEC) and high-performance liquid chromatography (HPLC). Brute mallein was also prepared to compare the results. The protein profile of the samples was analyzed using SDS-PAGE. The mallein test was also performed, and the results were evaluated using CFT. The AS method was selected as the optimal precipitation method. The protein profile exhibited a range of proteins from low to high molecular weights, appearing as a smear in the brute mallein. The mallein test using the AS-participated proteins, the first fraction from SEC, and the second fraction from HPLC yielded significant results, demonstrating erythema with a diameter of 18.46, 21.70 and 25.37 mm, respectively. The mallein test results were confirmed by CFT. The findings suggested that the purified antigenic proteins improved the mallein test and CFT results. Consequently, these proteins can be used to diagnose glanders correctly and increase the accuracy of the mallein test and CFT.

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Archives of Razi Institute

Articles in Press, Accepted Manuscript
Available Online from 11 November 2024

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