Improvement of recombinant-truncated Burkholderia motility protein A (BimA)-based indirect ELISA for equine glanders

Singh, Sandeep; Dohre, Sudhir K.; Kamthan, Aayushi; Pal, Vijai; Karothia, B S; Singha, Harisankar; Kumar, Subodh

doi:10.1080/15321819.2018.1523800

Cited by 6 publications

(3 citation statements)

References 15 publications

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“…Successful control of glanders infection requires reliable diagnostic tools. Although numerous methods [9][10][11]13,16,18,35] have been described for diagnosis of glanders, some have limitations [9][10][11]18]. Protein glycosylation, a posttranslational modification that covalently attaches a glycan to a protein, is prevalent in Archaea and bacteria [36,37].…”

Section: Discussionmentioning

confidence: 99%

“…A western blot technique based on a partially purified lipopolysaccharide-containing antigen shows higher specificity compared to CFT [14]. Moreover, indirect ELISAs based on the intracellular motility A protein (BimA) [15,16], the type 6 secreted TssB [17] and Hcp1 [18] proteins, the heat shock protein GroEL [19] and a semi-purified fraction of B. mallei have been developed [11]. However, these methods require more investigation using large-scale sample surveys and optimization to improve specificity and sensitivity [18].…”

Section: Introductionmentioning

confidence: 99%

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A glycoengineered antigen exploiting a conserved protein O-glycosylation pathway in the Burkholderia genus for detection of glanders infections

et al. 2021

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We recently described a protein O-glycosylation pathway conserved in all species of the Burkholderia genus that results in the synthesis and incorporation of a trisaccharide glycan to membrane-exported proteins. Here, we exploited this system to construct and evaluate a diagnostic tool for glanders. Burkholderia mallei, the causative agent of glanders, is a highly infectious and fatal zoonotic pathogen that infects horses, mules, donkeys, and occasionally humans. A highly sensitive and specific diagnostic tool is crucial for the control, elimination, and eradication of B. mallei infections. We constructed plasmids carrying synthetic genes encoding a modified, previously unannotated Burkholderia glycoprotein containing three glycosylation sequons fused to the cholera toxin B-subunit. The resulting proteins were glycosylated in the B. cenocepacia K56-2 parental strain, but not in glycosylationdeficient mutants, as determined by SDS-PAGE and fluorescent lectin blots. One of these glycoproteins was used as an antigen in ELISA and western blots to screen a panel of serum samples collected from glanders-infected and healthy horses, which were previously investigated by complement fixation test and indirect ELISA based on a semi-purified fraction of B. mallei. We show that ELISA and western blot assays based on our glycoprotein antigen provide 100% specificity, with a sensitivity greater than 88%. The glycoprotein antigen was recognized by serum samples collected from patients infected with B. pseudomallei, B. mallei, B. multivorans, and B. cenocepacia. Our results indicate that protein O-glycosylation in Burkholderia can be exploited as a biomarker for diagnosis of Burkholderiaassociated infections.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A glycoengineered antigen exploiting a conserved protein O-glycosylation pathway in the Burkholderia genus for detection of glanders infections

et al. 2021

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“…Other serologic assays, including ELISA, agglutination, and polysaccharide microarray, have been developed for glanders and have undergone various degrees of validation, but none are recommended by the WOAH over the CFT. 3,8,14,17,19,21,23 A commercial ELISA has been evaluated as a replacement for the CFT because it exhibited considerably fewer false-positives; ELISAs are well-suited for highthroughput laboratories. 4 A colorimetric western blot (WB) assay was developed as a highly specific confirmatory assay to resolve false-positive CFT results.…”

mentioning

confidence: 99%

Development and validation of a chemiluminescent western blot assay for glanders (Burkholderia mallei) serodetection

Grause,

Elschner,

Ledesma

et al. 2024

J VET Diagn Invest

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Glanders, caused by Burkholderia mallei, is a zoonotic disease of equids. Serologic testing for glanders is required by disease-free countries before international movement of equids. The World Organisation for Animal Health Terrestrial Manual recommends the complement fixation test (CFT) for clearance of individual animals for movement, but the CFT is prone to false-positive results. A colorimetric western blot (WB) assay was developed and validated to resolve false-positive CFT results; however, that assay is relatively time-consuming, and the interpretation is subjective. We present here a procedurally similar chemiluminescent WB assay that performs comparably to the validated colorimetric WB assay and offers noticeable benefits of decreased time-to-result and greater ease of interpretation.

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Glanders & Melioidosis - A Zoonosis and a Sapronosis

Virk

Fhogartaigh

Dance

2022

Zoonoses: Infections Affecting Humans and Animals

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Improvement of recombinant-truncated Burkholderia motility protein A (BimA)-based indirect ELISA for equine glanders

Cited by 6 publications

References 15 publications

A glycoengineered antigen exploiting a conserved protein O-glycosylation pathway in the Burkholderia genus for detection of glanders infections

A glycoengineered antigen exploiting a conserved protein O-glycosylation pathway in the Burkholderia genus for detection of glanders infections

Development and validation of a chemiluminescent western blot assay for glanders (Burkholderia mallei) serodetection

Glanders & Melioidosis - A Zoonosis and a Sapronosis

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