Dirofilaria immitis

Performance and standardization of specific antibody immunoassays for filariasis

Robert G Hamilton, Alan Leroy Scott, Robert D'Antonio, David A. Levy, N Franklin Adkinson

Research output: Contribution to journalArticle

Abstract

The factors associated with the development, optimization, and validation of immunoassays for the detection of parasite-specific antibody in filariasis infections were investigated using the dog heartworm, Dirofilaria immitis as a model. We examined two assays, the Protein A solid-phase radioimmunoassay (SPRIA) and enzyme-linked immunosorbent assay (ELISA), for quantitation of specific antibody to the parasite in canine serum. Precision, reproducibility, and parallelism were examined using response-error relationships and precision profile analyses. A staphylococcal Protein A saturation analysis was applied to the standardization of IgG antiparasite antibody reference sera in weight per volume units (μg/ml). Using the mean minimal detectable dose + 3 SD and an intraassay precision profile <10% coefficient of variation (CV) as criteria for assay sensitivity, the SPRIA and ELISA displayed comparable positive thresholds of 1 μg/ml IgG anti-parasite antibody/ml of serum. Both assays also demonstrated good reproducibility (<15% interassay CV) and parallelism (<20% interdilutional CV) over their working ranges (SPRIA: 1-40 μg/ml; ELISA: 1-5 μg/ml). Specificity of each assay was enhanced by preadsorption of cross-reacting antibodies in canine serum (i.e., specific for Toxocara canis antigens) with solid-phase antigen prior to assay. Methods for comparing different immunoassay designs are considered in relation to the variables that influence the assays' performance characteristics.

Original languageEnglish (US)
Pages (from-to)298-313
Number of pages16
JournalExperimental Parasitology
Volume56
Issue number3
DOIs
StatePublished - 1983

Fingerprint

Dirofilaria immitis
Filariasis
Immunoassay
Radioimmunoassay
Parasites
Antibodies
Enzyme-Linked Immunosorbent Assay
Staphylococcal Protein A
Serum
Canidae
Immunoglobulin G
Toxocara canis
Antigens
Anti-Idiotypic Antibodies
Weights and Measures
Infection

Keywords

  • antibody, quantitation
  • canine
  • Dirofilaria immitis
  • enzyme-linked immunosorbent assay (ELISA)
  • Filaria
  • immunoassay
  • nematodes, parasitic
  • precision profile analysis
  • quality control
  • saturation analysis
  • solid-phase radioimmunoassay (SPRIA)
  • Toxocara canis

ASJC Scopus subject areas

  • Immunology
  • Parasitology
  • Infectious Diseases

Cite this

Dirofilaria immitis : Performance and standardization of specific antibody immunoassays for filariasis. / Hamilton, Robert G; Scott, Alan Leroy; D'Antonio, Robert; Levy, David A.; Adkinson, N Franklin.

In: Experimental Parasitology, Vol. 56, No. 3, 1983, p. 298-313.

Research output: Contribution to journalArticle

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