Development of an immunomagnetic separation-ELISA for the detection of Burkholderia pseudomallei in blood samples.
Autor: Yatsomboon, Artitaya; Sermswan, Rasana W.; Wongratanacheewin, Surasakdi
Publication year: 2021
Asian Pacific journal of allergy and immunology
issn:0125-877X
doi: 10.12932/AP-080518-0307
Abstract:
BACKGROUND: Septicemic melioidosis caused by Burkholderia pseudomallei is a serious cause of morbidity and mortality. An effective, rapid and simple diagnostic method is required for detection of B. pseudomallei infection. OBJECTIVE: To develop immunomagnetic beads (IMB) coupled with ELISA (IMB-ELISA) for detection of B. pseudomallei in blood samples of patients with suspected melioidosis. METHODS: For separation of B. pseudomallei from buffer, blood samples and hemoculture, 200 nm immunomagnetic beads (IMBs) coated with 4B11 monoclonal antibody (4B11-IMBs) against exopolysaccharide antigens were used. The detection was done by an ELISA based biotin-streptavidin system. The sensitivity and specificity were evaluated. RESULTS: 4B11-IMBs (100 μg) were successfully developed and used for detection of B. pseudomallei in 1 ml samples. Transmission electron microscopy (TEM) imaging demonstrated B. pseudomallei was captured by 4B11-IMBs. The IMBs showed high capture efficiency (98%) with B. pseudomallei in buffer. The IMB-ELISA assay was highly specific for B. pseudomallei. It showed no cross-reactions with other bacteria, except B. mallei. The limits of the B. pseudomallei assay detection for detecting B. pseudomallei in either buffer solution or blood was 102 CFU/ml. The IMB-ELISA detection sensitivity in blood samples was 44.5%. Although it did not give the highest sensitivity, it was useful for detection with hemoculture that was faster than conventional methods. CONCLUSION: This study suggests the IMB-ELISA assay offers a simple and highly specific method with a turnaround time of 6 h for detection of B. pseudomallei. The developed assay can be applied in hospitals for surveillance of B. pseudomallei.
Language: eng
Rights:
Pmid: 30525739
Tags: Humans; Sensitivity and Specificity; Enzyme-Linked Immunosorbent Assay; Antibodies, Monoclonal/immunology; Antigens, Bacterial/immunology; Burkholderia pseudomallei/*immunology; Immunoglobulin G/immunology; Immunomagnetic Separation; Melioidosis/blood/*diagnosis; Polysaccharides, Bacterial/immunology; Sepsis/blood/*diagnosis
Link: https://pubmed.ncbi.nlm.nih.gov/30525739/
