title: Cloning, expression and purification of outer membrane protein (OmpA) of Burkholderia pseudomallei and evaluation of its potential for serodiagnosis of melioidosis.
abstract: 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.
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India.
authors.forename: Sonia
authors.initials: S
authors.lastname: Arora
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India. Electronic address: dtselvam@drde.drdo.in.
authors.forename: Duraipandian
authors.initials: D
authors.lastname: Thavaselvam
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India.
authors.forename: Ashu
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India.
authors.forename: Archana
authors.initials: A
authors.lastname: Prakash
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India.
authors.forename: Anita
authors.initials: A
authors.lastname: Barua
authors.affiliation: Division of Microbiology, Defence Research & Development Establishment, Jhansi Road, Gwalior 474 002 India.
authors.forename: Kannusamy
authors.initials: K
authors.lastname: Sathyaseelan
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D016957:Burkholderia pseudomallei / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D011994:Recombinant Proteins; C078167:OMPA outer membrane proteins
keywords: Burkholderia pseudomallei; ELISA; Melioidosis; OmpA; Serodiagnosis