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 language | English (US) |
|---|---|
| Pages (from-to) | 298-313 |
| Number of pages | 16 |
| Journal | Experimental Parasitology |
| Volume | 56 |
| Issue number | 3 |
| DOIs | |
| State | Published - 1983 |
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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 journal › Article
}
TY - JOUR
T1 - Dirofilaria immitis
T2 - Performance and standardization of specific antibody immunoassays for filariasis
AU - Hamilton, Robert G
AU - Scott, Alan Leroy
AU - D'Antonio, Robert
AU - Levy, David A.
AU - Adkinson, N Franklin
PY - 1983
Y1 - 1983
N2 - 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.
AB - 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.
KW - antibody, quantitation
KW - canine
KW - Dirofilaria immitis
KW - enzyme-linked immunosorbent assay (ELISA)
KW - Filaria
KW - immunoassay
KW - nematodes, parasitic
KW - precision profile analysis
KW - quality control
KW - saturation analysis
KW - solid-phase radioimmunoassay (SPRIA)
KW - Toxocara canis
UR - http://www.scopus.com/inward/record.url?scp=0021070271&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021070271&partnerID=8YFLogxK
U2 - 10.1016/0014-4894(83)90075-9
DO - 10.1016/0014-4894(83)90075-9
M3 - Article
VL - 56
SP - 298
EP - 313
JO - Experimental Parasitology
JF - Experimental Parasitology
SN - 0014-4894
IS - 3
ER -