title: Multiple-antigen ELISA for melioidosis--a novel approach to the improved serodiagnosis of melioidosis.
abstract: BACKGROUND
  Burkholderia pseudomallei, the causative agent of melioidosis, is endemic to Southeast Asia and northern Australia. Clinical manifestations of disease are diverse, ranging from chronic infection to acute septicaemia. The current gold standard of diagnosis involves bacterial culture and identification which is time consuming and often too late for early medical intervention. Hence, rapid diagnosis of melioidosis is crucial for the successful management of melioidosis.
  
  
  METHODS
  The study evaluated 4 purified B. pseudomallei recombinant proteins (TssD-5, Omp3, smBpF4 and Omp85) as potential diagnostic agents for melioidosis. A total of 68 sera samples from Malaysian melioidosis patients were screened for the presence of specific antibodies towards these proteins using enzyme-linked immunosorbent assay (ELISA). Sera from patients with various bacterial and viral infections but negative for B. pseudomallei, as well as sera from healthy individuals, were also included as non-melioidosis controls. The Mann Whitney test was performed to compare the statistical differences between melioidosis patients and the non-melioidosis controls.
  
  
  RESULTS
  TssD-5 demonstrated the highest sensitivity of 71% followed by Omp3 (59%), smBpF4 (41%) and Omp85 (19%). All 4 antigens showed equally high specificity (89-96%). A cocktail of all 4 antigens resulted in slightly reduced sensitivity of 65% but improved specificity (99%). Multiple-antigen ELISA provided improved sensitivity of 88.2% whilst retaining good specificity (96%). There was minimal reactivity with sera from healthy individuals proposing the utility of these antigens to demarcate diseased from non-symptomatic individuals in an endemic country.
  
  
  CONCLUSIONS
  TssD-5 demonstrated high detection sensitivity and specificity and the results were obtained within a few hours compared to time consuming culture and IFAT methods commonly used in a clinical setting. The use of multiple-antigens resulted in improved sensitivity (88.2%) whilst maintaining superior specificity. These data highlight the use of TssD-5 and other recombinant antigens tested in this study as potential serodiagnostic agents for melioidosis.
authors.affiliation: Malaysia Genome Institute, Jalan Bangi, Kajang, Selangor D.E. 43000, Malaysia. yukahara@gmail.com
authors.forename: Yuka
authors.initials: Y
authors.lastname: Hara
authors.forename: Chui-Yoke
authors.initials: CY
authors.lastname: Chin
authors.forename: Rahmah
authors.initials: R
authors.lastname: Mohamed
authors.forename: Savithri D
authors.initials: SD
authors.lastname: Puthucheary
authors.forename: Sheila
authors.initials: S
authors.lastname: Nathan
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial; D016957:Burkholderia pseudomallei / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008554:Melioidosis / Q000175:diagnosis*; D008875:Middle Aged; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins
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