Serologic Responses to the Mallein Test for Glanders in Solipeds

Hagebock, Joyce M.; Schlater, Linda K.; Frerichs, Wayne M.; Olson, David P.

doi:10.1177/104063879300500121

Cited by 26 publications

(27 citation statements)

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“…A redução na quantidade de anticorpos, observada nos animais 60 dias após a inoculação com a B. mallei inativada provavelmente está relacionada ao período de soroconversão causado pelas proteínas desse agente que é de no máximo dois meses (Hagebock et al 1993). Entretanto, exposições constantes de animais às proteínas do agente podem promover uma soroconversão permanente (Gregory & Waag 2008).…”

Section: Resultsunclassified

Desenvolvimento e avaliação de um teste ELISA indireto para o diagnóstico sorológico do mormo em equídeos

et al. 2012

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Glanders is an infectious-contagious disease of acute or chronic character which principally affects horses, causing enormous losses in the productive chain of this animal. To control the disease, the Ministry of Agriculture, Husbandry and Supply instituted mandatory sanitation measures in the entire national territory which include an ofϐicial diagnosis through the complement ϐixation (CF) test, maleinization and sacriϐice of the animals that are positive. Nowadays the kits used for the diagnosis of the disease are imported, making their routine application difϐicult and more expensive. The objective of this study was to standardize an indirect ELISA test, using the proteic extract of Burkholderia mallei isolated from a carrier horse in the state of Pernambuco. The samples were cultivated in 10% blood agar and incubated for 48h at 37°C; later, one of the isolated colonies was characterized phenotypically and genotypically and immediately cultivated in brain heart infusion (BHI) for enrichment; then it was peaked (repicada) for the Dor-set Henley medium which was incubated at 37ºC under 60rpm for eight weeks. To standardize the test the Protein G Peroxidase Sigma Conjugate was used in the dilution of 1:90.000, with serums diluted in 1:100 and the antigen in 1:400. Sixty serums were used as negative controls, tested before the CF to determine the cutting point which was 0.042nm. After establishing the standardization, 300 samples were tested, of which 99% (297) were in agreement with the results obtained in the CF. At the end, of assay presented 100% sensibility and 98.2% speciϐicity, with predictive (preditivo) positive and negative values of 97.7% and 100% respectively. The Kappa concordance test was 0.98 and the intra and interplac repeatability were 8.8% and 10.3% respectively. From the results obtained, it is possible to afϐirm that the indirect ELISA test can be used as an efϐicient diagnosis tool. However, more essays must be carried out to consolidate the reliability of this test.

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Section: Resultsunclassified

Desenvolvimento e avaliação de um teste ELISA indireto para o diagnóstico sorológico do mormo em equídeos

et al. 2012

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“…Mallein, a heat-killed, primarily lipopolysaccharide (LPS) extract of B. mallei, was used as the glanders cELISA antigen. 7,13 Both piroplasmosis antigens, the dourine antigen, and the glanders antigen were stored at Ϫ70 C until used.…”

Section: Antigensmentioning

confidence: 99%

“…6,7,9,11,12 This report describes a single, rapid, simple protocol for the competitive ELISA (cELISA) of equine serum designed to replace the current CF procedures for the serodiagnosis of piroplasmosis, dourine, and glanders.…”

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confidence: 99%

“…2,3 These CF procedures do not function with sera having anticomplementary activity, test interpretation is often subjective, test sensitivity is low relative to more modern immunoassay methods, and CF test sensitivity declines as the serologic responses of exposed animals shift from initial IgMbased reactions to those of other immunoglobulin classes and subclasses. 6,7,9,11,12 This report describes a single, rapid, simple protocol for the competitive ELISA (cELISA) of equine serum designed to replace the current CF procedures for the serodiagnosis of piroplasmosis, dourine, and glanders.…”

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confidence: 99%

“…2,3 The OIE has also recognized alternative serodiagnostic methods for each etiologic agent, including separate indirect fluorescent antibody tests (IFATs) for B. equi and B. caballi, enzyme-linked immunosorbent assay (ELISA) for B. mallei, and ELISA, IFAT, or agar gel immunodiffusion procedures for T. equiperdum serodiagnosis. 4,6,7,12,13,17 In practice, the battery of 4 CF procedures has been the most convenient, rapid, single test system for accomplishing the required piroplasmosis, dourine, and glanders serodiagnostic testing. Unfortunately, the CF tests require careful continuous titration of numerous labile reagents and, for piroplasmosis, the laborious expensive preparation of antigen from infected splenectomized horses.…”

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confidence: 99%

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Procedurally Similar Competitive Immunoassay Systems for the Serodiagnosis ofBabesia Equi, Babesia Caballi, Trypanosoma Equiperdum, andBurkholderia MalleiInfection in Horses

2000

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Abstract. Procedurally similar competitive enzyme-linked immunoassay (cELISA) methods were developed for the serodiagnosis of Babesia equi and Babesia caballi (piroplasmosis), Trypanosoma equiperdum (dourine), and Burkholderia mallei (glanders) infections in horses. Apparent test specificities for the B. equi, B. caballi, T. equiperdum, and B. mallei cELISAs were 99.2%, 99.5%, 98.9%, and 98.9%, respectively. Concordances and kappa values between the complement fixation (CF) and the cELISA procedures for the serodiagnosis of B. equi, B. caballi, T. equiperdum, and B. mallei infections in experimentally exposed horses were 76% and 0.55, 89% and 0.78, 97% and 0.95, and 70% and 0.44, respectively. The cELISA method may be a technically more reproducible, objective, and convenient approach for piroplasmosis, dourine, and glanders serodiagnosis in qualifying animals for international movement and disease eradication programs than the CF systems currently in use. Use of the cELISA method also obviated the problems associated with testing hemolyzed or anticomplementary sera.Piroplasmosis, dourine, and glanders are internationally reportable, serious infectious diseases of Equidae. 1,3 The etiologic agents of these diseases are unrelated. Piroplasmosis results from Babesia equi or Babesia caballi infection, whereas dourine and glanders result from Trypanosoma equiperdum and Burkholderia mallei infections respectively. 1,3 Many nations require that Equidae presented for importation be serologically negative to B. equi, B. caballi, T. equiperdum, and B. mallei. 1,3,10 Traditionally, a panel of 4 separate complement fixation (CF) procedures has been the primary serodiagnostic tool prescribed by the Office International des Epizooties (OIE) for this purpose. 2,3 The OIE has also recognized alternative serodiagnostic methods for each etiologic agent, including separate indirect fluorescent antibody tests (IFATs) for B. equi and B. caballi, enzyme-linked immunosorbent assay (ELISA) for B. mallei, and ELISA, IFAT, or agar gel immunodiffusion procedures for T. equiperdum serodiagnosis. 4,6,7,12,13,17 In practice, the battery of 4 CF procedures has been the most convenient, rapid, single test system for accomplishing the required piroplasmosis, dourine, and glanders serodiagnostic testing. Unfortunately, the CF tests require careful continuous titration of numerous labile reagents and, for piroplasmosis, the laborious expensive preparation of antigen from infected splenectomized horses. 2,3 These CF procedures do not From the Diagnostic Bacteriology Laboratory, National Veterinary Services Laboratories, US Department of Agriculture, Animal and Health Inspection Services, Ames, IA 50010.Received for publication January 19, 1999. function with sera having anticomplementary activity, test interpretation is often subjective, test sensitivity is low relative to more modern immunoassay methods, and CF test sensitivity declines as the serologic responses of exposed animals shift from initial IgMbased reactions to those of other immu...

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