title: Potential immunogenic polypeptides of Burkholderia pseudomallei identified by shotgun expression library and evaluation of their efficacy for serodiagnosis of melioidosis.
abstract: The search for novel immunogenic polypeptides to improve the accuracy and reliability of serologic diagnostic methods for Burkholderia pseudomallei infection is ongoing. We employed a rapid and efficient approach to identify such polypeptides with sera from melioidosis patients using a small insert genomic expression library created from clinically confirmed local virulent isolates of B. pseudomallei. After 2 rounds of immunoscreening, 6 sero-positive clones expressing immunogenic peptides were sequenced and their identities were: benzoate 1,2-dioxygenase beta subunit, a putative 200 kDa antigen p200, phosphotransferase enzyme family protein, short chain dehydrogenase and 2 hypothetical proteins. These immunogens were then transferred to an ELISA platform for further large scale screening. By combining shotgun expression library and ELISA assays, we identified 2 polypeptides BPSS1904 (benzoate 1,2-dioxygenase beta subunit) and BPSL3130 (hypothetical protein), which had sensitivities of 78.9% and 79.4% and specificities of 88.1% and 94.8%, respectively in ELISA test, thus suggesting that both are potential candidate antigens for the serodiagnosis of infections caused by B. pseudomallei.
authors.affiliation: Department of Biomedical Science, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Suat Moi
authors.initials: SM
authors.lastname: Puah
authors.forename: S D
authors.initials: SD
authors.lastname: Puthucheary
authors.forename: Kek Heng
authors.initials: KH
authors.lastname: Chua
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D016957:Burkholderia pseudomallei / Q000276:immunology* / Q000378:metabolism / Q000472:pathogenicity; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D006801:Humans; D008297:Male; D008554:Melioidosis / Q000276:immunology / Q000378:metabolism / Q000382:microbiology*; D010105:Oxygenases / Q000096:biosynthesis / Q000302:isolation & purification; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D010770:Phosphotransferases / Q000096:biosynthesis / Q000302:isolation & purification; D012698:Serologic Tests; D012703:Serotyping
chemical_list: D001426:Bacterial Proteins; D010455:Peptides; D010105:Oxygenases; C019766:benzoate 1,2-dioxygenase; D010770:Phosphotransferases
keywords: ELISA.; melioidosis; recombinant polypeptide; serodiagnosis; shotgun expression library