Comparison of Immunogenicity of 17 Burkholderia Mallei Antigens and Whole Cell Lysate Using Indirect ELISA

Glanders is a World Organization for Animal Health (WOAH)-notifiable equine disease caused by the infection of Burkholderia mallei, and is endemic in Mongolia, South Asia, Africa, and South America. While the complement fixation test (CFT) has been widely used for serodiagnosis of glanders and is considered a standard serological test, it has several limitations. These limitations include poor specificity, labor intensive techniques, variability in antigen and protocol. Consequently, indirect enzyme-linked immunosorbent assays (iELISAs) based on recombinant proteins have been developed as alternative serodiagnostic assays to address some of the challenges associated with the CFT. The accuracy of iELISA relies on the B. mallei proteins used as an antigen. Hence, to determine the best diagnostic candidate in iELISA, in terms of sensitivity and specificity, a comparison of 17 immunogenic B. mallei proteins and detergent-based whole cell lysate (WCL) was performed. According to the sensitivity and specificity on the sera from glanderous and non-glanderous Mongolian native horses, iELISA using Hcp1, GroEL, and detergent-based WCL represented the highest diagnostic accuracy. These three candidates did not have cross-reactivity to horse sera with several other equine diseases. WCL, Hcp1, and GroEL showed considerable potential as antigens for iELISA in the serodiagnosis of glanders in Mongolia. Detergent-based WCL extraction offers a consistent approach for the preparation of reliable B. mallei antigen. WCL-iELISA should be further validated in a large-scale study to meet WOAH demands.