Identification of a new diagnostic antigen for glanders using immunoproteome analysis

Dohre, Sudhir K.; Kamthan, Aayushi; Singh, Sandeep; Alam, Syed Imteyaz; Kumar, Subodh

doi:10.1016/j.cimid.2017.06.007

Cited by 13 publications

(11 citation statements)

References 26 publications

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“…mallei were also evaluated for use in ELISA, e.g. the intracellular motility A protein (BimA) [14], the type 6 secreted protein TssB [18] and a heat shock protein GroEL [21]. A new indirect ELISA based on a semi-purified fraction of B .…”

Section: Introductionmentioning

confidence: 99%

Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders

Elschner¹,

Laroucau²,

Singha³

et al. 2019

PLoS ONE

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Glanders is a zoonotic contagious disease of equids caused by Burkholderia (B.) mallei . Serodiagnosis of the disease is challenging because of false-positive and false-negative test results. The accuracy of the complement fixation test (CFT) which is prescribed for international trade by the World Organisation for Animal Health (OIE), five ELISAs and a Western blot (WB) were compared for serodiagnosis of glanders using sera from 3,000 glanders-free and 254 glanderous equids. Four ELISA tests are based on recombinant antigens (TssA, TssB, BimA and Hcp1), the IDVet ELISA is based on a semi-purified fraction of B . mallei and WB makes use of a purified LPS-containing B . mallei -antigen. Sensitivity and specificity of tests were estimated using cut-off values recommended by the test developers. The WB and all ELISAs, except BimA, were significantly more specific than the CFT. ELISAs based on TssA, TssB, and BimA antigens had significantly lower sensitivity compared to CFT while the sensitivities of the Hcp1-ELISA, the IDVet-ELISA and the WB did not differ significantly from that of the CFT. Given their comparable sensitivities and specificities, the CFT (98.0%, 96.4%), the WB (96.8%, 99.4%), the Hcp1-ELISA (95.3%, 99.6%) and the IDVet-ELISA (92.5%, 99.5%) should be further developed to meet OIE requirements.

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Section: Introductionmentioning

confidence: 99%

Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders

Elschner¹,

Laroucau²,

Singha³

et al. 2019

PLoS ONE

Self Cite

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“…The supernatant was precipitated with tricarboxylic acid (TCA) (Merck, Germany). The washed pellet was resuspended in PBS (0.01 M, pH 7.2) (5). Total protein concentration was evaluated using the Bradford method by measuring the absorbance at 595 nm (UV/Vis spectrophotometer 6320D, Jenway, UK) using bovine serum albumin (BSA) as the standard.…”

Section: Methodsmentioning

confidence: 99%

“…Generally, glanders in horse tends to be a chronic disease but in other equidae (i.e., mule, zebra, and donkey) the disease has an acute nature (3, 4). In equids, glanders is transmitted by oro-nasal mucous membrane invasion and inhalation routes (5). The common symptoms of glanders include nasal discharge (mucopurulent), lung lesions and nodules involving the liver and spleen (3).…”

Section: Introductionmentioning

confidence: 99%

“…The serological tests, including complement fixation test (CFT), enzyme-linked immunosorbent assays (ELISA), Rose Bengal plate agglutination test (RBPT), indirect fluorescent antibody test (IFAT), indirect hemagglutination test (IHAT), and counter immunoelectrophoresis test (CIET) have been used for diagnosis of glanders where there is technically no flawless method of choice (14). CFT however, is currently the most widely used test in international trade of equines and is recommended by the World Organisation of Animal Health (OIE) (12) though some limitations such as false positive/negative results have been noticed (5, 15, 16). The CFT is a labor-intensive and time-consuming test with sensitivity of 90–95%, and a questionable specificity (5).…”

Section: Introductionmentioning

confidence: 99%

“…CFT however, is currently the most widely used test in international trade of equines and is recommended by the World Organisation of Animal Health (OIE) (12) though some limitations such as false positive/negative results have been noticed (5, 15, 16). The CFT is a labor-intensive and time-consuming test with sensitivity of 90–95%, and a questionable specificity (5). There is, therefore, room for development of an immunoblotting-based method providing high specificity.…”

Section: Introductionmentioning

confidence: 99%

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Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm

Yazdansetad

Mosavari

Tadayon

et al. 2019

IJM

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Background and Objectives: Burkholderia mallei is the leading cause of glanders, a highly transmittable and an OIE-no- tifiable disease of equidae. Despite the importance of B. mallei, little is known about serodiagnosis of glanders. The present study aimed to develop an immunoblotting assay based on whole-cell proteome of B. mallei to enable accurate serodiagnosis of glanders. Materials and Methods: Three farm horses were subcutaneously immunized with a crude suspension (106 cfu/ml) of heat-inactivated B. mallei formulated with incomplete Freund's adjuvant (IFA) to achieve a hyperimmune sera panel. The immunization was done for 1, 14 and 28 days with 1 dose of 1 ml antigen containing 106 cfu/ml. The hyperimmunity of sera was confirmed by CFT. B. mallei whole-cell proteome was prepared through sonication and the protein content was visu- alized by SDS-PAGE and quantified by Western blot using HRP-conjugated rabbit anti-horse IgG. A comprehensive set of positive and negative horse sera validated the test. Results: A ladder pattern of the B. mallei immunoreactive antigens was seen within the region of 20-90 kDa clearly and the immunoblot was scored positive, while no reaction was seen for the negative sera. The Western blot assay indicated a noticeably higher diagnostic specificity for positive or negative sera of glanders. Conclusion: The whole-cell proteome-based immunoblot proved reliable and straightforward in our study. The prepared an- tigen was adaptable for application in immunoblotting. We assumed this improved immunoblotting system provides appro- priate sensitivity and also specificity expected in serodiagnosis of glanders in endemic areas and typically in less-developed countries.

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Development of a microsphere‐based immunoassay for the serological detection of glanders in equids

et al. 2020

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Identification of a new diagnostic antigen for glanders using immunoproteome analysis

Cited by 13 publications

References 26 publications

Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders

Evaluation of the comparative accuracy of the complement fixation test, Western blot and five enzyme-linked immunosorbent assays for serodiagnosis of glanders

Development of an immunoblotting assay for serodiagnosis of Burkholderia mallei infection: the whole-cell proteome-based paradigm

Development of a microsphere‐based immunoassay for the serological detection of glanders in equids

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