Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Abstract

Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e., crude preparations of whole cells. False-positive results obtained from other diagnostic tests utilizing crude antigens lead to financial losses to animal owners, and false-negative results can turn a risk into a possible threat. In this study, we report on the identification of diagnostic targets using bioinformatics tools for serodiagnosis of glanders. The identified gene sequences were cloned and expressed as recombinant proteins. The purified recombinant proteins of B. mallei were used in an indirect ELISA format for serodiagnosis of glanders. Two recombinant proteins, 0375H and 0375TH, exhibited 100% sensitivity and specificity for glanders diagnosis. The proteins also did not cross-react with sera from patients with the closely related disease melioidosis. The results of this investigation highlight the potential of recombinant 0375H and 0375TH proteins in specific and sensitive diagnosis of glanders.

Development of a real-time loop-mediated isothermal amplification assay for detection ofBurkholderia mallei

“…Burkholderia mallei ‐specific target DNA sequence was selected as described earlier (Pal, Kumar, Malik, & Rai, ; Pal, Singh, Tiwari, Jaiswal, & Rai, ). A subregion (89 bp) of gene BMA10229_0375 along with 145‐bp downstream sequence of BMA10229_0375 being specific for B. mallei was targeted for development of LAMP assay.…”

“…To the best of our knowledge, this is the first report of a specific and sensitive real-time LAMP assay for detection of B. mallei. 2.2 | Selection of B. mallei-specific target DNA sequence Burkholderia mallei-specific target DNA sequence was selected as described earlier (Pal, Kumar, Malik, & Rai, 2012;Pal, Singh, Tiwari, Jaiswal, & Rai, 2016). A subregion (89 bp…”

A Quadruplex Real-Time PCR Assay for the Rapid Detection and Differentiation of the Most Relevant Members of the B. pseudomallei Complex: B. mallei, B. pseudomallei, and B. thailandensis

“…A gene that encodes a 16.5 kDa hypothetical protein (Accession number YP_001024199 Locus tag = BMA10229_0375) was reported to be unique to B . mallei [ 77 ]. Orf11 (Accession number AF074878) was reported to be unique to B .…”

Seroprevalence of equine glanders in horses in the central and eastern parts of Mongolia

“…However, due to animal welfare concerns, this test is not currently recommended [26]. The complement fixation test (CFT) is considered a suitable screening test for the diagnosis of equine glanders [4,11,12,25,27,28,30].…”