Recombinant truncated flagellin of Burkholderia pseudomallei as a molecular probe for diagnosis of melioidosis.
Current serological tests for melioidosis, using impure or uncharacterized cell antigens from Burkholderia pseudomallei, have problems in detection sensitivity and specificity. Therefore, we designed and expressed the recombinant flagellin (truncated at both the N- and C-terminal ends), and used the antigen to develop an indirect enzyme-linked immunosorbent assay (ELISA) to diagnose melioidosis. Comparison of the immunoreactivities of the full-length and truncated flagellins reveals that the truncated flagellin performed much better in detection specificity and sensitivity. Only the full-length flagellin was recognized by other bacterial causing septicemia and gave a false-positive result in Western analysis, indicating that the cross-reactive epitopes were located on the more highly conserved N- and C-terminal regions of flagellin. The indirect ELISA using recombinant truncated flagellin as the antigen achieved 93.8% sensitivity and 96.3% specificity and offered a more efficient serodiagnosis of melioidosis.
Section of Infectious Disease, Kaohsiung Veterans General Hospital, Taiwan, Republic of China.
Yao-Shen
YS
Chen
David
D
Shiuan
Ssu-Ching
SC
Chen
Soi-Moi
SM
Chye
Ya-Lei
YL
Chen
D000942:Antigens, Bacterial / Q000235:genetics; D016957:Burkholderia pseudomallei / Q000276:immunology* / Q000302:isolation & purification; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards; D000939:Epitopes; D005408:Flagellin / Q000235:genetics*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008554:Melioidosis / Q000175:diagnosis*; D015335:Molecular Probes; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity; D017384:Sequence Deletion; D012698:Serologic Tests / Q000379:methods / Q000592:standards
D000942:Antigens, Bacterial; D000939:Epitopes; D015335:Molecular Probes; D011994:Recombinant Proteins; D005408:Flagellin