title: Potential of recombinant flagellin fragment from Burkholderia thailandensis as an antigen for melioidosis antibody detection by indirect ELISA.
abstract: Non-pathogenic Burkholderia thailandensis may be used as a model for Burkholderia pseudomallei due to the genetic similarity of these species. Moreover, the experimental manipulation of B. thailandensis is safer. In this study, we constructed recombinant flagellin protein fragments of B. thailandensis E264 (FLAG300, FLAG600, FLAG900, and FLAGFL fragments) and used fragments as the antigen to detect melioidosis antibodies by an indirect enzyme-linked immunosorbent assay (indirect ELISA). The serum samples consisted of serodiagnostic melioidosis sera (N = 52), septicemic sera caused by other bacteria (disease control) (N = 16) and healthy donor sera (N = 40). The area under the receiver operating characteristic (ROC) curve at optimal value (mean plus standard deviation) of all constructed fragments to melioidosis antibodies detection ranged from 0.654 to 0.953. The highest area under the ROC curve (AUROCC, 0.953) for FLAG300 fragment at 1600 serum titer revealed the best performance of melioidosis diagnosis. The indirect ELISA using this fragment as the antigen possessed 82.7% sensitivity and 94.6% specificity.
authors.affiliation: Biochemistry Unit, Department of Medical Sciences, Faculty of Science, Rangsit University, Muang Ake, Pathumthain 12000, Thailand. sumetw@hotmail.com
authors.forename: Sumet
authors.initials: S
authors.lastname: Wajanarogana
authors.forename: Phitchapat
authors.initials: P
authors.lastname: Nimnuch
authors.forename: Acharawan
authors.initials: A
authors.lastname: Thongmee
authors.forename: Kanyanan
authors.initials: K
authors.lastname: Kritsiriwuthinan
mesh_terms: D000907:Antibodies, Bacterial / Q000032:analysis / Q000097:blood; D000942:Antigens, Bacterial / Q000276:immunology; D001483:Base Sequence; D019117:Burkholderia / Q000235:genetics* / Q000472:pathogenicity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005408:Flagellin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis* / Q000382:microbiology; D008969:Molecular Sequence Data; D012372:ROC Curve; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins; D005408:Flagellin