A statistical model to optimize enzyme-linked immunosorbent assay parameters for detection of Mycoplasma gallisepticum and M. synoviae antibodies in egg yolk.

H. O. Mohammed, R. Yamamoto, Tim Carpenter, H. B. Ortmayer

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

An enzyme-linked immunosorbent assay (ELISA) was developed to quantify Mycoplasma gallisepticum (MG) and M. synoviae (MS) antibodies in egg-yolk extract. Various parameters of ELISA were evaluated and optimized. A statistical model was developed to study the relationship between ELISA absorbance (A) and antigen concentration, antibody concentration, and time of reading of the test. These factors explained 62% of the variability in A for the MG antigen and 74% of the variability in A for the MS antigen. The optimum concentration of MG antigen was 4 micrograms protein/ml, and that for MS antigen was 3 micrograms protein/ml. A similar model was developed to study the relation between A and conjugate concentration, antibody concentration, and time of reading of the test. The optimum concentration of the conjugate was found to be 1:4000. The within-run and between-run coefficients of variation for MG were 8% and 9%, respectively, and those for MS were 9% and 7%, respectively, indicating a high degree of reproducibility of these tests. There was a high correlation between ELISA and hemagglutination-inhibition test results.

Original languageEnglish (US)
Pages (from-to)389-397
Number of pages9
JournalAvian Diseases
Volume30
Issue number2
DOIs
StatePublished - Jan 1 1986
Externally publishedYes

ASJC Scopus subject areas

  • Food Animals
  • Animal Science and Zoology
  • Immunology and Microbiology(all)

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