Isolation and Purification of theAntigenic Proteins and its Use in Diagnostic Tests.

Abstract

Glanders is a contagious infectious disease caused bythat affects both solipeds and carnivores. This disease occasionally leads to human infection through direct contact between humans and infected animals. The recent rise in glanders prevalence has intensified focus on control and eradication programs, emphasizing accurate diagnosis of infected cases using high-performance test. To this end, antigenic proteins were purified from Burkholderia mallei, and the mallein test was optimized using these purified proteins. Finally, the efficacy of antigenic proteins was evaluated using the complement fixation test (CFT). The laboratory strain ofwas selected,and proteins from 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 for comparison. The protein profile of the samples was analyzed using SDS-PAGE. The mallein test was also performed, and results were evaluated using CFT. The AS method was identified 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. Mallein test using AS-participated proteins, the first SEC fraction and the second HPLC fraction yielded significant results, demonstrating erythema diameters of 18.46, 21.70 and 25.37 mm, respectively. These mallein test results were confirmed by CFT. The findings indicated that the purified antigenic proteins improved both mallein test and CFT results. Consequently, these proteins can diagnose glanders correctly and increase the accuracy of the mallein test and CFT.