Development of an avidin-biotin dot enzyme-linked immunosorbent assay and its comparison with other serological tests for diagnosis of glanders in equines

Verma, R.D.; Sharma, Jitendra Kumar; Venkateswaran, K.; Batra, Harsh Vardhan

doi:10.1016/0378-1135(90)90095-d

Cited by 33 publications

(22 citation statements)

References 12 publications

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“…Mesmo havendo uma ótima concordância do ELISA-i quando comparado com o teste de ϐixação de complemento (FC) que possui sensibilidade em torno dos 90% a 95%, este último tem o risco de apresentar reações falso-negativas ocasionalmente observadas no soro de alguns animais (jovens, gestantes ou idosos), além de reações falso-positivas que podem ocorrer em aproximadamente 1% dos soros testados ao se utilizar antígeno de células inteiras, o que o leva a ser superado pelos resultados obtidos no ELISA-i (Cravitz & Miller 1950, Verma et al 1990. O animal gestante que apresentou resultado positivo no ELISA e negativo na Fixação de Complemento, provavelmente deve-se à limitação desse úlƟ mo teste, que pode apresentar resultados falsos-negativos, especialmente em soros de burros, mulas e animais gestantes (Gregory & Waag, 2008).…”

Section: Resultsunclassified

“…Além disso, existem entraves quanto a padronização insuϐiciente do antígeno, o que pode comprometer a qualidade da reação, resultando em limitações no que diz respeito à especiϐicidade e sensibilidade da técnica. Alguns pesquisadores ainda questionam a eϐicácia e aplicabilidade do referido ensaio (Jana et al 1982, Verma et al 1990, Neubauer et al 2005, Naureen et al 2007.…”

Section: Introductionunclassified

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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

Section: Introductionunclassified

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

et al. 2012

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“…(Svobodova et al, 2013). It was reported that the dot-ELISA is simple, quick, specific, sensitive, low cost field test that detects minute levels of antibodies much faster than complement fixation test and indirect hemagglutination antibody test (Verma & Misra, 1989;Verma et al, 1990). Dot-ELISA has been used for the serodiagnosis of glanders (John et al, 2010).…”

Section: Enzyme-linked Immunosorbant Assay (Elisa)mentioning

confidence: 99%

Biotechnological tools for diagnosis of equine infectious diseases

Prasad¹,

Brar²,

Ikbal³

et al. 2016

JEBAS

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Rapid diagnosis of infectious diseases and appropriate treatment with in time are important steps that promote optimal clinical outcomes and general public health. Today there is large number of new technologies such as nanotechnology, biosensors, and microarray techniques, are being developed and used as diagnostic tools for equine infectious diseases. Nucleic acid based techniques such as polymerase chain reaction (PCR) have become conventional tools in veterinary research and plays an important role in specific typing determinations as well as for rapid screening of ample numbers of samples at the time of equine disease outbreaks. Other biotechnological techniques are populous to be used in the coming times as they can enhance diagnostic efficacy in less time and cost as compared to conventional techniques. This review focuses on biotechnological tools available for equine diseases diagnosis and its applications hold great promise for improving the speed and accuracy of diagnostics for equine infectious diseases.

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“…B. mallei usually spread via lymphatics. The isolation of bacillus from nasal discharge is not easy as it is usually highly contaminated Several other serological tests were developed which include agglutination test [12], Indirect haemagglutination (IHA) test [13], precipitation test [12], immunodiffusion test [14], counter-immuno-electrophoresis test [15], [17], enzyme-linked immunosor-bent assay (ELISA) [18].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of PCR, DNA hybridization and immunomagnetic separation — PCR for detection of Burkholderia mallei in artificially inoculated environmental samples

et al. 2010

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Development of an avidin-biotin dot enzyme-linked immunosorbent assay and its comparison with other serological tests for diagnosis of glanders in equines

Cited by 33 publications

References 12 publications

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

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

Biotechnological tools for diagnosis of equine infectious diseases

Evaluation of PCR, DNA hybridization and immunomagnetic separation — PCR for detection of Burkholderia mallei in artificially inoculated environmental samples

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