Cloning, expression and purification of outer membrane protein (OmpA) of Burkholderia pseudomallei and evaluation of its potential for serodiagnosis of melioidosis

• Cloning, expression, and purification of recombinant protein OmpA of B. pseudomallei.
• Standardization of iELISA with rOmpA.
• Evaluation in indirect ELISA format with 87 serum samples.
• Assay sensitivity of 82.6% and specificity of 93.75%.
• iELISA will be a useful tool in serodiagnosis of melioidosis.

Melioidosis is an emerging infectious disease in India and caused by gram-negative, soil saprophyte bacteria Burkholderia pseudomallei. This disease is endemic in Southeast Asia and northern Australia, and sporadic cases of melioidosis are also reported from southern states of India. The present study reports the cloning, expression, and purification of recombinant protein outer membrane protein A (OmpA) of B. pseudomallei and its evaluation in indirect enzyme-linked immunosorbent assay (ELISA) format with 87 serum samples collected from Manipal, Karnataka, India. Twenty-three samples from culture confirmed cases (n = 23) of melioidosis, 25 serum samples from patients of other febrile illness and pyrexia of unknown origin (n = 25), and 39 serum samples from healthy blood donors (n = 39) from Kasturba Medical College, Manipal, were tested in this assay format. The assay showed sensitivity of 82.6% and specificity of 93.75%. The recombinant OmpA based indirect ELISA will be a useful tool for serodiagnosis of melioidosis in large scale rapid screening of clinical samples.