Development and validation of an indirect enzyme immunoassay for detection of ovine antibody to Brucella ovis

Vigliocco, Ana M.; Paulo, Patricia Silva; Mestre, J.; Briones, Gabriel; Draghi, Graciela; Tossi, Mirta; Nielsen, Klaus

doi:10.1016/s0378-1135(96)01285-0

Cited by 32 publications

(19 citation statements)

References 20 publications

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“…23 For the evaluation of repeatability, the CV values of 13.4% for the equivocal sera and 0.64% for the positive sera, are considered as acceptable and comparable to those reported by other authors of 5.98-11.8%. 3,4,17 Although SD values for reproducibility have not, to our knowledge, been reported, the value obtained for the cELISA can be considered satisfactory because the value is similar to the values obtained by other authors for repeatability.…”

Section: 22supporting

confidence: 81%

Validation according to OIE criteria of a monoclonal, recombinant p26–based, serologic competitive enzyme-linked immunosorbent assay as screening method in surveillance programs for the detection of Equine infectious anemia virus antibodies

et al. 2016

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Abstract. The Italian National Reference Center for equine infectious anemia (CRAIE; Rome, Italy) developed and validated a monoclonal, recombinant p26-based competitive enzyme-linked immunosorbent assay (cELISA) for the detection of EIA virus antibodies employing the 2010 criteria of the World Organization for Animal Health (OIE). The following parameters were evaluated: cutoff values, repeatability, reproducibility, concordance, analytical sensitivity (Se), absolute analytical specificity (Sp), and diagnostic Se and Sp. Positive and negative predictive values were also defined in relation to the estimated prevalence. When the cELISA was used as a screening test for 96,468 samples in the Italian EIA surveillance program, 17% more EIA cases were detected than by the agar gel immunodiffusion test, and the apparent diagnostic Sp estimated from these samples was 99.8%, which was more than the diagnostic Sp (80.2%) estimated from validation. The high Se and Sp of the cELISA confirm its fit for purpose as a screening test.

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Section: 22supporting

confidence: 81%

Validation according to OIE criteria of a monoclonal, recombinant p26–based, serologic competitive enzyme-linked immunosorbent assay as screening method in surveillance programs for the detection of Equine infectious anemia virus antibodies

et al. 2016

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“…A confirmação do diagnósti-co pode ser feita pelo cultivo em meios seletivos para isolamento da bactéria a partir do sêmen de carneiros, descargas vaginais e leite das ovelhas (Bulgin 1990, Grilló et al 1999, Baigún et al 2000. No entanto, os métodos indiretos são amplamente utilizados, sendo a imunodifusão em gel de ágar (IDGA), a reação de fixação do complemento (RFC) e o ensaio imunoenzimático indireto (ELISA) os métodos sorológicos mais utilizados em vários países (Burgess & Norris 1982, Worthington et al 1984, Vigliocco et al 1997, Robles 1998 animais (IBGE 1998). Nas mesorregiões do Agreste e Mata Paraibana a pecuária é menos expressiva e a agricultura é bastante diversificada, predominando os pequenos estabelecimentos agropecuários.…”

Section: Introdução Introdução Introdução Introdução Introduçãounclassified

Inquérito soro-epidemiológico e fatores de risco associados à infecção por Brucella ovis em carneiros deslanados do semi-árido da Paraíba

et al. 2007

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Foi realizado um levantamento soro-epidemiológico da brucelose por Brucella ovis em reprodutores ovinos deslanados na Paraíba com os objetivos de verificar a prevalência e distribuição da infecção por B. ovis em propriedades rurais e analisar os possíveis fatores de risco associados à infecção. Foram investigadas 283 propriedades criadoras de ovinos, das quais foram colhidas 498 amostras de soro sanguíneo de carneiros, a partir de oito meses de idade, nas mesorregiões do Sertão Paraibano e Borborema. Todos os soros foram examinados pela IDGA (teste de triagem) e RFC (teste confirmatório). De acordo com as análises, 8,59% (I.C.95% = 5,83%-12,48%) das propriedades apresentaram evidência sorológica de infecção por B. ovis, com uma prevalência de 5,57% (I.C.95% = 3,86%-7,97%) de reprodutores sororreagentes. Nas propriedades que higienizavam suas instalações com maior freqüência, a soropositividade foi estatisticamente inferior (p < 0,05).

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“…However, CFT has some drawbacks like complexity, incompatibility with anticomplementary and hemolyzed sera, prozone phenomena and occasional false reactions. On the other hand, AGID is a very labour intensive test and has low number of test capacity [2][3][4]6,18,19] . Of the serological methods used to detect antibodies to B. ovis, I-ELISA has been shown to be the most sensitive and specific test.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, among chronically infected rams which show CFT negative results are not rare. On the other hand, AGID is a very labour intensive test and has low sample capacity [2][3][4][5][6] . Various I-ELISAs have been developed for detection of antibodies against B. ovis antigens with various results.…”

Section: Introductionmentioning

confidence: 99%

Brucella abortus RB51 ve Brucella canis M- Varyant Suşlarından Hazırlanan Antijenlerin Kullanıldığı Enzim Bağlı İmmünosorbent Testinin Koyunlarda Brucella ovis İnfeksiyonunun Serolojik Tanısında Kullanılabilirliğinin Araştırılması

Tel¹,

Keskin²

2015

Kafkas Univ Vet Fak Derg

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The aim of this work was to investigate the possible usage of hot saline extract antigens (HSE) of Brucella abortus RB51(HSE-RB51) and B. canis M-variant strains (HSE-M-) in ELISA by comparing the results with those of obtained from commercial I-ELISA kit for the serological diagnosis of B. ovis infection. In this study, a total of 183 serum samples collected from different cities (Şanlıurfa, Mardin, Gaziantep, Diyarbakır) in Southeastern Anatolia Region of Turkey were tested by using three ELISAs, one of which is from a commercial source, Rose Bengal plate agglutination tests prepared by rough (R-RBPT) and smooth strains (S-RBPT). Recombinant protein A/G conjugated with horseradish peroxidase (A/G-HRPO) was used as conjugate in the in house ELISAs. Seropositivity rate was 11% for HSE-RB51-ELISA and 3.3% for both HSE-M-and commercial ELISA. The percentage of positive results was 7.6% for S-RBPT and 2.7% for R-RBPT. Only 2 serum samples were positive for all the tests except S-RBPT. Because similar results were obtained from the same serum samples by both commercial ELISA and HSE-M-ELISA, these results may suggest that HSE-M-antigen could be used in ELISA for serologic diagnosis of B. ovis infection in sheep. Since 7.6% of the serum samples were found as positive by only HSE-RB51-ELISA, it was assumed that this test could be less specific or more sensitive than other tests used in the study. Although R-RBPT is a screening test, it showed the lowest seropositivity in the study. This could be explained by less mucoid nature of its antigen than other test systems using natural rough species. S-RBPT ile serumların %7.6'sı ve R-RBPT ile %2.7'si pozitif bulundu. Sadece 2 serum R-RBPT ve 3 ELISA ile pozitif reaksiyon verdi. Ticari B. ovis ELISA kiti ile STE-M-ELISA ile aynı sayıda ve aynı serumlardan pozitif reaksiyon alınması STE-M-antijeninin B. ovis infeksiyonunun serolojik tanısında ELISA için iyi bir aday antijen olabileceğinin kanısına varıldı. Kullanılan serolojik testlerden sadece STE-RB51-ELISA ile pozitiflik yüzdesinin %7.6 olması testin özgüllüğünün diğer testlere göre daha düşük olabileceği kanısı uyandırdı. Ayrıca R-RBPT' nin bir tarama testi olmasına rağmen en düşük seropozitiflik oranı göstermesinin M-varyant suşunun daha az mukoid bir yapı içermesi nedeni ile çapraz reaksiyonları azaltması sonucu olabileceği düşünüldü. Keywords

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Development and validation of an indirect enzyme immunoassay for detection of ovine antibody to Brucella ovis

Cited by 32 publications

References 20 publications

Validation according to OIE criteria of a monoclonal, recombinant p26–based, serologic competitive enzyme-linked immunosorbent assay as screening method in surveillance programs for the detection of Equine infectious anemia virus antibodies

Validation according to OIE criteria of a monoclonal, recombinant p26–based, serologic competitive enzyme-linked immunosorbent assay as screening method in surveillance programs for the detection of Equine infectious anemia virus antibodies

Inquérito soro-epidemiológico e fatores de risco associados à infecção por Brucella ovis em carneiros deslanados do semi-árido da Paraíba

Brucella abortus RB51 ve Brucella canis M- Varyant Suşlarından Hazırlanan Antijenlerin Kullanıldığı Enzim Bağlı İmmünosorbent Testinin Koyunlarda Brucella ovis İnfeksiyonunun Serolojik Tanısında Kullanılabilirliğinin Araştırılması

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