Use of a Western blot technique for the serodiagnosis of glanders

Elschner, Mandy C.; Scholz, Holger C.; Melzer, Falk; Saqib, Muhammad; Marten, Peggy; Rassbach, Astrid; Dietzsch, Michael; Schmoock, Gernot; Santana, Vania Lucia de Assis; Souza, Marcília Maria Alves de; Wernery, Renate; Wernery, Ulrich; Neubauer, Heinrich

doi:10.1186/1746-6148-7-4

Cited by 42 publications

(69 citation statements)

References 21 publications

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“…Approaches using polysaccharide-based microarray technology (23) are expensive and will not be suitable for routine mass testing of serum samples. Recently, an LPS-based Western blot technique with 100% sensitivity and specificity has been described for serodiagnosis of glanders (10). Preparation of LPS is sophisticated and each time of preparation requires culture handling in a BSL3 laboratory.…”

Section: Discussionmentioning

confidence: 99%

“…Other serological tests, e.g., agglutination, precipitation, indirect hemagglutination, immunodiffusion, counterimmunoelectrophoresis, and enzyme-linked immunosorbent assay (ELISA), have also been described (6,13,26,33), but they have limitations. A Western blot technique using lipopolysaccharide antigen has been shown to have higher sensitivity and specificity than the currently used CFT (10).…”

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confidence: 99%

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Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Pal

Kumar

Malik

et al. 2012

Clin. Vaccine Immunol.

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Glanders is a contagious disease caused by the Gram-negative bacillus Burkholderia mallei. The number of equine glanders outbreaks has increased steadily during the last decade. The disease must be reported to the Office International des Epizooties, Paris, France. Glanders serodiagnosis is hampered by the considerable number of false positives and negatives of the internationally prescribed tests. The major problem leading to the low sensitivity and specificity of the complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA) has been linked to the test antigens currently used, i.e., crude preparations of whole cells. False-positive results obtained from other diagnostic tests utilizing crude antigens lead to financial losses to animal owners, and false-negative results can turn a risk into a possible threat. In this study, we report on the identification of diagnostic targets using bioinformatics tools for serodiagnosis of glanders. The identified gene sequences were cloned and expressed as recombinant proteins. The purified recombinant proteins of B. mallei were used in an indirect ELISA format for serodiagnosis of glanders. Two recombinant proteins, 0375H and 0375TH, exhibited 100% sensitivity and specificity for glanders diagnosis. The proteins also did not cross-react with sera from patients with the closely related disease melioidosis. The results of this investigation highlight the potential of recombinant 0375H and 0375TH proteins in specific and sensitive diagnosis of glanders.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Pal

Kumar

Malik

et al. 2012

Clin. Vaccine Immunol.

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“…The CFT as the recommended test is also subject to some limitations, such as the possibility of false-positive results. A Western immunoblot technique has recently been proposed for resolving the false-positives found in the CFT for use in areas with low glanders prevalence (Elschner et al 2011). …”

Section: Discussionmentioning

confidence: 99%

Tying-up syndrome associated with glanders in two mules

Araghi-Sooreh¹,

Mahmoudi²,

Bahmani³

et al. 2015

PHK

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Two mules with lameness and tight hind limbs were referred to the veterinary teaching hospital due to losing weight and honeycoloured nasal discharge. Other clinical findings included increased rectal temperature, emaciation and increased heart rate (> 80 bpm). Blood samples were taken for haematological and serum biochemical examinations. Creatine kinase (CK) and aspartate transaminase (AST) enzymes showed considerable increases in both cases. The mules were checked using the intradermopalpebral mallein test, and glanders was diagnosed for both cases. To the best of our knowledge, there have been no reports published of tying-up syndrome associated with glanders. Regarding the clinical signs and the results of the increased CK and AST enzymes and myoglobinuria, the most plausible cause of the tying-up syndrome was considered to be rhabdomyolysis. Localisation of the bacteria into the muscles after bacteremia, and subsequent myositis or sedimentation of antigen-antibody complexes in muscles followed by autoimmune reactions may be involved in the clinical signs reported here.

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“…Although diagnostic tests have been developed for use in equines (4,9,13), no serological tests have been developed to identify glanders in humans. Skin tests using mallein did not become positive in horses until 3 to 4 weeks after infection and therefore may have little value diagnostically (10).…”

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confidence: 99%

Humoral Immune Responses in a Human Case of Glanders

Waag¹,

England²,

DeShazer³

2012

Clin. Vaccine Immunol.

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ABSTRACTWithin 2 months of acquiring glanders, a patient developed 8-, 16-, and 4-fold increases, respectively, in specific IgA, IgG, and IgM serological titers againstBurkholderia mallei. Within 14 months of infection, the titers decreased to the baseline. Serum from this patient was also highly reactive againstBurkholderia pseudomalleiwhole cells.Burkholderia malleiwhole cells did not react with sera from patients with other diseases. Therefore, an assay using aB. malleicellular diagnostic antigen may be useful for the serodiagnosis of glanders.

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Use of a Western blot technique for the serodiagnosis of glanders

Cited by 42 publications

References 21 publications

Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Evaluation of Recombinant Proteins of Burkholderia mallei for Serodiagnosis of Glanders

Tying-up syndrome associated with glanders in two mules

Humoral Immune Responses in a Human Case of Glanders

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