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Recombinant Truncated Flagellin of Burkholderia pseudomallei as a Molecular Probe for Diagnosis of Melioidosis

Section of Infectious Disease, Kaohsiung Veterans General Hospital,¹ Institute of Biomedical Science, National Sun Yat-Sen University,² Department of Medical Technology, Fooyin Unversity, Kaohsiung,⁵ Department of Life Science and Institute of Biotechnology, National Dong-Hwa University, Hualien,³ Department of Medicine, China Medical Collage, Taichung, Taiwan, Republic of China⁴

Received 22 October 2002/ Returned for modification 6 January 2003/ Accepted 4 February 2003

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.