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Evaluation of recombinant antigens for diagnosis of melioidosis
Elizabeth M Allwood1, Carie-Anne Logue, Gregory J Hafner
1Institute for Glycomics, Griffith University Gold Coast Campus, Qld, Australia.
Abstract:
Burkholderia pseudomallei, the causative agent of melioidosis, is endemic to Southeast Asia and northern Australia. Clinical manifestations of the disease are diverse, ranging from chronic localized infection to acute septicaemia, with death occurring within 24-48 h after the onset of symptoms. Definitive diagnosis of melioidosis involves bacterial culture and identification, with results obtained within 3-4 days. This delayed diagnosis is a major contributing factor to high mortality rates. Rapid diagnosis is vital for successful management of the disease. This study describes the purification and evaluation of three recombinant antigenic proteins, BPSL0972, BipD and OmpA from B. pseudomallei 08, for their potential in the serodiagnosis of melioidosis using an indirect enzyme-linked immunosorbent assay (ELISA) method. The recombinant proteins were evaluated using 74 serum samples from culture-confirmed melioidosis patients from Malaysia, Thailand and Australia. In addition, 62 nonmelioidosis controls consisting of serum samples from clinically suspected melioidosis patients (n=20) and from healthy blood donors from an endemic region (n=18) and a nonendemic region (n=24) were included. The indirect ELISAs using BipD and BPSL0972 as antigens demonstrated poor to moderate sensitivities (42% and 51%, respectively) but good specificity (both 100%). In contrast, the indirect ELISA using OmpA as an antigen achieved 95% sensitivity and 98% specificity. These results highlight the potential for OmpA to be used in the serodiagnosis of melioidosis in an endemic area.
Insights
Rapid serodiagnosis of melioidosis is crucial for patient survival. This study evaluated recombinant proteins for diagnosing melioidosis, finding OmpA highly effective with 95% sensitivity and 98% specificity.
Area of Science:
- Medical Microbiology
- Immunodiagnostics
- Infectious Diseases
Background:
- Melioidosis, caused by Burkholderia pseudomallei, is prevalent in Southeast Asia and Australia.
- Diverse clinical presentations range from chronic localized infections to acute septicemia, often with rapid mortality.
- Current definitive diagnosis relies on bacterial culture, taking 3-4 days, contributing to high mortality rates.
Purpose of the Study:
- To evaluate three recombinant antigenic proteins (BPSL0972, BipD, OmpA) from B. pseudomallei for their potential in serodiagnosis of melioidosis.
- To assess the diagnostic accuracy of these proteins using an indirect enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- Purification and preparation of recombinant BPSL0972, BipD, and OmpA proteins.
- Indirect ELISA was performed on 74 serum samples from culture-confirmed melioidosis patients.
- Control groups included serum from suspected melioidosis patients and healthy donors from endemic and non-endemic regions.
Main Results:
- Indirect ELISAs using BipD and BPSL0972 showed poor to moderate sensitivity (42% and 51%) but 100% specificity.
- The indirect ELISA employing OmpA achieved a high sensitivity of 95% and specificity of 98%.
- OmpA demonstrated significant potential for accurate melioidosis serodiagnosis.
Conclusions:
- OmpA is a promising candidate antigen for the development of a rapid and accurate serodiagnostic test for melioidosis.
- Improved diagnostic tools are essential for timely intervention and reducing mortality associated with melioidosis in endemic areas.
