Evaluation of recombinant proteins of Burkholderia mallei for serodiagnosis of glanders.
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.
Division of Microbiology, Defence Research and Development Establishment, Gwalior, Madhya Pradesh, India. vjsoni@rediffmail.com
Vijai
V
Pal
Subodh
S
Kumar
Praveen
P
Malik
Ganga Prasad
GP
Rai
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D042726:Burkholderia mallei / Q000235:genetics / Q000276:immunology*; D019295:Computational Biology / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005896:Glanders / Q000175:diagnosis*; D006736:Horses; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D014730:Veterinary Medicine / Q000379:methods*
D000942:Antigens, Bacterial; D011994:Recombinant Proteins