A Sensitive Serodiagnostic Test for Onchocerciasis Using a Cocktail of Recombinant Antigens

Bradley, Joshua; Trenholme, Katharine R.; Gillespie, A. J.; Guderian, Ronald H.; Titanji, V. P. K.; Hong, Yan; McReynolds, Larry A.

doi:10.4269/ajtmh.1993.48.198

Cited by 57 publications

(54 citation statements)

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“…Serology -To detect prepatent or cryptic infections an ELISA-based assay, which used a cocktail of 3 recombinant antigens show to have a high specificity and sensitivity was performed (Bradley et al 1993). Finger-prick capillary samples were collected on filter paper, dried, and stored at -20 o C before testing.…”

Section: Study Area -mentioning

confidence: 99%

“…The final concentration was 2 µg/ml in the proportion of 50% Ov 11, 25% Ov 10, and 25% Ov 29 (Ov 10 and Ov 29, 500 ng/ml; and Ov 11, 1 µg/ml). The ELISA methodology, which has been reported previously (Bradley et al 1991(Bradley et al , 1993, was performed with the following modifications. The plates were blocked for 1 hr with 5% nonfat skim milk in PBS and serum samples were added at 1/200 dilution in 0.05% Tween 20 (PBS-T) with 5% nonfat skim milk.…”

Section: Study Area -mentioning

confidence: 99%

“…The plates were developed with 2,2'-azino-di-(ethyl benzathiozoline) substrate (Sigma, St.Louis, MO). A positive test was determined as previously described (Bradley et al 1993).…”

Section: Study Area -mentioning

confidence: 99%

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Onchocerciasis in Ecuador: Prevalence of Infection on the Ecuador-Colombia Border in the Province of Esmeraldas

Guderian¹,

Anselmi²,

Espinel³

et al. 1997

Mem. Inst. Oswaldo Cruz

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Section: Study Area -mentioning

confidence: 99%

Section: Study Area -mentioning

confidence: 99%

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Onchocerciasis in Ecuador: Prevalence of Infection on the Ecuador-Colombia Border in the Province of Esmeraldas

Guderian¹,

Anselmi²,

Espinel³

et al. 1997

Mem. Inst. Oswaldo Cruz

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“…Além disso, Madruga et al (2000) Outro ponto importante nesse trabalho foi a padronização de um ELISA utilizando como antígeno um coquetel de proteínas recombinantes. A hipótese inicial era de que uma maior variedade de epítopos resultaria em um aumento significativo da sensibilidade do teste, conforme observado por Bradley et al (1993) para Onchocerca volvulus. Porém, o mesmo não foi observado para A. marginale.…”

Section: Resultsunclassified

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA

et al. 2010

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Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática-ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI). Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.

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“…[8][9][10][11][12][13][14] Some of these antigens were tested in a multicenter trial organized by the Tropical Diseases Research Program of the WHO that identified highly specific proteins, 15 and high sensitivity of these tests was achieved by combining three proteins in a diagnostic cocktail. 13,16,17 However, these antigens were difficult to produce and the fact that they were expressed as fusions with a carrier protein made it necessary to run parallel tests to rule out antibody reactions with the carrier. These limitations have prevented the O. volvulus antigen cocktail from becoming a widely used diagnostic tool.…”

Section: Introductionmentioning

confidence: 99%

Sensitive and specific serodiagnosis of onchocerciasis with recombinant hybrid proteins.

Nde¹,

Pogonka²,

Bradley³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. Onchocerciasis remains a major health hazard in many tropical countries. However, the existing tools for diagnosis of the disease have limitations, particularly regarding the detection of low level or early infections. To design an optimized reagent, we exploited the high antibody reactivity of patient sera against the Onchocerca volvulus proteins Ov20 and Ov33, which have been described as highly sensitive and specific immunodiagnostic reagents for producing hybrid proteins. The construct OvH2 was composed of Ov20 fused to Ov33, while OvH3 consisted of the C-terminus of Ov20 linked to Ov33. When these constructs were tested with sera from patients with onchocerciasis and control sera, OvH2 showed a sensitivity of 98.5% and a specificity of 97.7% and OvH3 showed a sensitivity of 98.5% and a specificity of 95.35%. All non-responders were from Ecuador. These results surpass those of existing single recombinant antigens, suggesting that our hybrid proteins combined the sensitivity of the two parent proteins. Tests based on OvH2 should prove suitable for monitoring onchocerciasis control programs and individual diagnosis.

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A Sensitive Serodiagnostic Test for Onchocerciasis Using a Cocktail of Recombinant Antigens

Cited by 57 publications

References 0 publications

Onchocerciasis in Ecuador: Prevalence of Infection on the Ecuador-Colombia Border in the Province of Esmeraldas

Onchocerciasis in Ecuador: Prevalence of Infection on the Ecuador-Colombia Border in the Province of Esmeraldas

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA

Sensitive and specific serodiagnosis of onchocerciasis with recombinant hybrid proteins.

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