Towards single screening tests for brucellosis

K. Nielsen; P. Smith; W. Yu; P. Nicoletti; P. Elzer; C. Robles; R. Bermudez; T. Renteria; F. Moreno; A. Ruiz; C. Massengill; Q. Muenks; Gregers Jungersen; T. Tollersrud; L. Samartino; S. Conde; L. Forbes; D. Gall; B. Perez; X. Rojas; A. Minas

Towards single screening tests for brucellosis

K. Nielsen, P. Smith, W. Yu, P. Nicoletti, P. Elzer, C. Robles, R. Bermudez, T. Renteria, F. Moreno, A. Ruiz, C. Massengill, Q. Muenks, Gregers Jungersen, T. Tollersrud, L. Samartino, S. Conde, L. Forbes, D. Gall, B. Perez, X. RojasA. Minas

Research output: Contribution to journal › Journal article › Research › peer-review

Abstract

This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLIPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.

Original language	English
Journal	Revue Scientifique Et Technique-office International Des Epizooties
Volume	24
Issue number	3
Pages (from-to)	1027-1037
ISSN	0253-1933
Publication status	Published - 2005

Keywords

indirect enzyme-linked immunosorbent assay
universal assay
fluorescence polarisation assay
diagnosis
brucellosis
serology

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Nielsen, K., Smith, P., Yu, W., Nicoletti, P., Elzer, P., Robles, C., Bermudez, R., Renteria, T., Moreno, F., Ruiz, A., Massengill, C., Muenks, Q., Jungersen, G., Tollersrud, T., Samartino, L., Conde, S., Forbes, L., Gall, D., Perez, B., ... Minas, A. (2005). Towards single screening tests for brucellosis. Revue Scientifique Et Technique-office International Des Epizooties, 24(3), 1027-1037.

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title = "Towards single screening tests for brucellosis",

abstract = "This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLIPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.",

keywords = "indirect enzyme-linked immunosorbent assay, universal assay, fluorescence polarisation assay, diagnosis, brucellosis, serology",

author = "K. Nielsen and P. Smith and W. Yu and P. Nicoletti and P. Elzer and C. Robles and R. Bermudez and T. Renteria and F. Moreno and A. Ruiz and C. Massengill and Q. Muenks and Gregers Jungersen and T. Tollersrud and L. Samartino and S. Conde and L. Forbes and D. Gall and B. Perez and X. Rojas and A. Minas",

year = "2005",

language = "English",

volume = "24",

pages = "1027--1037",

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Nielsen, K, Smith, P, Yu, W, Nicoletti, P, Elzer, P, Robles, C, Bermudez, R, Renteria, T, Moreno, F, Ruiz, A, Massengill, C, Muenks, Q, Jungersen, G, Tollersrud, T, Samartino, L, Conde, S, Forbes, L, Gall, D, Perez, B, Rojas, X & Minas, A 2005, 'Towards single screening tests for brucellosis', Revue Scientifique Et Technique-office International Des Epizooties, vol. 24, no. 3, pp. 1027-1037.

TY - JOUR

T1 - Towards single screening tests for brucellosis

AU - Nielsen, K.

AU - Smith, P.

AU - Yu, W.

AU - Nicoletti, P.

AU - Elzer, P.

AU - Robles, C.

AU - Bermudez, R.

AU - Renteria, T.

AU - Moreno, F.

AU - Ruiz, A.

AU - Massengill, C.

AU - Muenks, Q.

AU - Jungersen, Gregers

AU - Tollersrud, T.

AU - Samartino, L.

AU - Conde, S.

AU - Forbes, L.

AU - Gall, D.

AU - Perez, B.

AU - Rojas, X.

AU - Minas, A.

PY - 2005

Y1 - 2005

N2 - This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLIPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.

AB - This paper describes an indirect enzyme-linked immunosorbent assay (I-ELISA) and a fluorescence polarisation assay (FPA), each capable of detecting antibody in several species of hosts to smooth and rough members of the genus Brucella. The I-ELISA uses a mixture of smooth lipopolysaccharide (SLPS) and rough lipopolysaccharide (RLPS) as the antigen, and a recombinant protein A/G conjugated with horseradish peroxidase as the detection reagent. When using individually determined cutoff values, the SLPS/RLPS combined-antigen I-ELISA detected antibody in slightly more animals exposed to SLPS or to RLPS than did I-ELISA procedures using each individual antigen separately. Similarly, the assay using combined antigens detected antibody in slightly fewer animals not exposed to Brucella sp. When a universal cutoff of 10% positivity was used (relative to strongly positive control sera of each species), the overall performance index (percentage sensitivity plus percentage specificity) value decreased by 1.0 (from 199.4 to 198.4). In the FPA, it was not possible to use a universal cutoff without significant loss of performance. The overall sensitivity value for the FPA using the combined FPA antigen was 1.0% lower than using the O-polysaccharide (OPS) from SLIPS and 9.1% higher than using the core antigen (CORE) from RLPS. When the combined antigen was used, the FPA specificity was slightly higher (1.2%) than from only the OPS, and considerably higher (12.6%) than the CORE. Overall, both the I-ELISA and the FPA with combined antigens were suitable as screening tests for all species of Brucella in the animal species tested.

KW - indirect enzyme-linked immunosorbent assay

KW - universal assay

KW - fluorescence polarisation assay

KW - diagnosis

KW - brucellosis

KW - serology

M3 - Journal article

SN - 0253-1933

VL - 24

SP - 1027

EP - 1037

JO - Revue Scientifique Et Technique-office International Des Epizooties

JF - Revue Scientifique Et Technique-office International Des Epizooties

IS - 3

ER -

Towards single screening tests for brucellosis

Abstract

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