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Evaluation of a Radioimmunoassay for Type III Procollagen Peptide in the Dog
Author(s) -
Dimski Donna S.,
Brooks Charles L,
Johnson Susan E.
Publication year - 1990
Publication title -
veterinary clinical pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.537
H-Index - 51
eISSN - 1939-165X
pISSN - 0275-6382
DOI - 10.1111/j.1939-165x.1990.tb00541.x
Subject(s) - radioimmunoassay , antiserum , serial dilution , polyacrylamide gel electrophoresis , microbiology and biotechnology , peptide , medicine , chemistry , endocrinology , biology , antibody , biochemistry , immunology , pathology , enzyme , alternative medicine
Summary Type III procollagen peptide (P‐3‐P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P‐3‐P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P‐3‐P, followed by Western immunoblotting with rabbit anti‐P‐3‐P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P‐3‐P and sera from two dogs suspected of having elevated P‐3‐P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross‐reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P‐3‐P. All tested sera from dogs had minimal competitive binding with radiolabeled P‐3‐P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P‐3‐P concentrations in any of the groups of dogs studied. It was concluded that currently available radioimmunoassay kits for the measurement of P‐3‐P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P‐3‐P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.