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Proteomic-based identification of novel EV-derived protein antibodies biomarkers for melioidosis diagnosis
Nini Luo1, Jun Tan1, Xuemiao Li1
1NHC Key Laboratory of Tropical Disease Control, School of Life Sciences and Medical Technology, Hainan Medical University, Haikou, Hainan, China.
Abstract:
Melioidosis, caused by Burkholderia pseudomallei (Bp), is a life-threatening disease characterized by diverse clinical manifestations and limited diagnostic capabilities. Extracellular vesicles (EVs) have emerged as critical carriers of novel antibody targets for serodiagnosis. In this study, we established a Bp-infected BEAS-2B cell model (Bp/BEAS-2B) and isolated EV from both Bp and Bp/BEAS-2B cells to generate EV proteome, identifying potential antigenic biomarkers for melioidosis diagnosis. Bioinformatics analysis identified PPEP and POMCR proteins as candidate antigens, with BLF1 and omp A serving as positive controls. Using a self-developed IgM-ELISA, serum samples from 43 melioidosis patients and 47 healthy volunteers were analyzed to detect antibodies against these antigens. Anti-POMCR IgM demonstrated exceptional diagnostic performance, with an AUC of 0.9872 (95% CI: 0.9713-1.003), sensitivity of 93.02% and specificity of 97.92% at a cutoff value of OD450 = 0.118. Similarly, IgM against PPEP, BLF1, and omp A also showed high diagnostic accuracy, with AUC values of 0.969, 0.9621, and 0.976, respectively. The accuracy of anti-POMCR and anti-PPEP were 96.43% and 95.54%, respectively, equivalent to anti-omp A (93.75%) and anti-BLF1 (91.96%). Antibodies to EV-derived proteins effectively differentiated melioidosis patients from other bacterial infections and healthy volunteers, highlighting their clinical potential as diagnostic tools for melioidosis.
Insights
This study identifies novel antibody targets from extracellular vesicles for diagnosing melioidosis. Antibodies against POMCR and PPEP proteins show high accuracy, offering potential for improved melioidosis diagnostics.
Area of Science:
- Microbiology
- Immunology
- Biotechnology
Background:
- Melioidosis, a severe infection caused by Burkholderia pseudomallei (Bp), presents diagnostic challenges due to varied symptoms and limited tests.
- Extracellular vesicles (EVs) are increasingly recognized as sources of biomarkers for disease diagnosis.
- Identifying specific antibody targets is crucial for developing accurate serodiagnostic tools for melioidosis.
Purpose of the Study:
- To identify novel antigenic biomarkers from Bp-derived extracellular vesicles (EVs) for melioidosis serodiagnosis.
- To evaluate the diagnostic performance of antibodies against identified EV proteins in melioidosis patients.
- To assess the potential of these EV-derived antigens as diagnostic tools for melioidosis.
Main Methods:
- Established a Burkholderia pseudomallei-infected BEAS-2B cell model (Bp/BEAS-2B).
- Isolated EVs from Bp and Bp/BEAS-2B cells to generate EV proteome.
- Utilized bioinformatics to identify candidate antigens (PPEP, POMCR) and positive controls (BLF1, omp A).
- Developed an IgM-Enzyme-Linked Immunosorbent Assay (ELISA) to detect antibodies in serum samples from melioidosis patients and healthy volunteers.
Main Results:
- Anti-POMCR IgM antibodies showed high diagnostic performance with an Area Under the Curve (AUC) of 0.9872, 93.02% sensitivity, and 97.92% specificity.
- Antibodies against PPEP, BLF1, and omp A also demonstrated high diagnostic accuracy (AUCs ranging from 0.9621 to 0.976).
- The accuracy for anti-POMCR and anti-PPEP was 96.43% and 95.54%, respectively, comparable to positive controls.
Conclusions:
- EV-derived proteins, particularly POMCR and PPEP, serve as effective antigenic biomarkers for melioidosis diagnosis.
- Antibodies targeting these EV-derived proteins can accurately differentiate melioidosis patients from healthy individuals and those with other bacterial infections.
- These findings highlight the significant clinical potential of EV-derived antibody targets for developing improved melioidosis diagnostic tools.
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