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An improved ELISA system for the measurement of IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in human sera.
Dev Biol Stand. 1991; 73:167-74.DB

Abstract

We have developed a modified assay method for the measurement of antibodies against PT and FHA. Polystyrene balls coated with purified PT and FHA were incubated with standard or test sera, washed, incubated with goat anti-human IgG F(ab')2 conjugated with horseradish peroxidase and then washed again. Peroxidase activity was then measured by color development using o-phenylenediamine. The anti-PT and anti-FHA IgG titers were determined using calibration curves. The assay standard curves for anti-PT and anti-FHA were both linear from 1 to 100 ELISA units per milliliter (EU ml-1), and the assay sensitivity was calculated to be 1 EU ml-1 in both cases. Intra- and inter-assay CVs were both less than 10%. Inter-laboratory assay CVs calculated from the results from four different facilities were 3.3-34.6% (average 14.5%) and 2.1-30.3% (average 13.6%) for anti-PT and anti-FHA IgG, respectively, and the correlation coefficient between the Chinese hamster ovary (CHO) cell assay and the anti-PT PS-ball ELISA assay was 0.959. Using this assay method we obtained in a highly sensitive and reproducible measurement of IgG antibodies against PT and FHA in human sera. This assay should be especially useful for the determination of low concentrations of these antibodies.

Authors+Show Affiliations

Department of Bacteriology and Applied Immunology, National Institute of Health, Tokyo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

1778310

Citation

Sato, Y, et al. "An Improved ELISA System for the Measurement of IgG Antibodies Against Pertussis Toxin (PT) and Filamentous Hemagglutinin (FHA) in Human Sera." Developments in Biological Standardization, vol. 73, 1991, pp. 167-74.
Sato Y, Sato H, Kodama H, et al. An improved ELISA system for the measurement of IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in human sera. Dev Biol Stand. 1991;73:167-74.
Sato, Y., Sato, H., Kodama, H., Uchimura, M., Miwa, N., Kobayashi, T., Yamamoto, E., Fujita, I., & Kumamoto, T. (1991). An improved ELISA system for the measurement of IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in human sera. Developments in Biological Standardization, 73, 167-74.
Sato Y, et al. An Improved ELISA System for the Measurement of IgG Antibodies Against Pertussis Toxin (PT) and Filamentous Hemagglutinin (FHA) in Human Sera. Dev Biol Stand. 1991;73:167-74. PubMed PMID: 1778310.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An improved ELISA system for the measurement of IgG antibodies against pertussis toxin (PT) and filamentous hemagglutinin (FHA) in human sera. AU - Sato,Y, AU - Sato,H, AU - Kodama,H, AU - Uchimura,M, AU - Miwa,N, AU - Kobayashi,T, AU - Yamamoto,E, AU - Fujita,I, AU - Kumamoto,T, PY - 1991/1/1/pubmed PY - 1991/1/1/medline PY - 1991/1/1/entrez SP - 167 EP - 74 JF - Developments in biological standardization JO - Dev Biol Stand VL - 73 N2 - We have developed a modified assay method for the measurement of antibodies against PT and FHA. Polystyrene balls coated with purified PT and FHA were incubated with standard or test sera, washed, incubated with goat anti-human IgG F(ab')2 conjugated with horseradish peroxidase and then washed again. Peroxidase activity was then measured by color development using o-phenylenediamine. The anti-PT and anti-FHA IgG titers were determined using calibration curves. The assay standard curves for anti-PT and anti-FHA were both linear from 1 to 100 ELISA units per milliliter (EU ml-1), and the assay sensitivity was calculated to be 1 EU ml-1 in both cases. Intra- and inter-assay CVs were both less than 10%. Inter-laboratory assay CVs calculated from the results from four different facilities were 3.3-34.6% (average 14.5%) and 2.1-30.3% (average 13.6%) for anti-PT and anti-FHA IgG, respectively, and the correlation coefficient between the Chinese hamster ovary (CHO) cell assay and the anti-PT PS-ball ELISA assay was 0.959. Using this assay method we obtained in a highly sensitive and reproducible measurement of IgG antibodies against PT and FHA in human sera. This assay should be especially useful for the determination of low concentrations of these antibodies. SN - 0301-5149 UR - https://www.unboundmedicine.com/medline/citation/1778310/An_improved_ELISA_system_for_the_measurement_of_IgG_antibodies_against_pertussis_toxin__PT__and_filamentous_hemagglutinin__FHA__in_human_sera_ DB - PRIME DP - Unbound Medicine ER -