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Development of a fecal sample collection strategy for extraction and quantification of fecal immunoglobulin A in dogs

Ursula Tress Dr med vet1, Jan S. Suchodolski Dr med vet, PhD2, David A. Williams VetMB, PhD3, and Jörg M. Steiner Dr med vet, PhD4
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  • 1 Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843-4474.
  • | 2 Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843-4474.
  • | 3 Department of Veterinary Clinical Medicine, University of Illinois, Urbana, IL 61802.
  • | 4 Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843-4474.

Abstract

Objective—To develop a fecal sample collection strategy and quantification method for measurement of fecal IgA concentrations in dogs.

Sample Population—Fecal samples from 23 healthy pet dogs of various breeds.

Procedures——Immunoglobulin A was extracted from fecal samples. An ELISA for the measurement of fecal IgA concentrations was established and analytically validated. Intraindividual variation of fecal IgA was determined by calculation of coefficients of variation. A sample collection strategy was developed on the basis of results of intraindividual variation of fecal IgA concentrations. A reference range for fecal IgA concentrations was determined.

Results—The method for extraction and quantification of fecal IgA was determined to be sufficiently sensitive, reproducible, accurate, and precise. On the basis of the intraindividual variability of our results, the determined fecal sample collection strategy required analysis of a total of 4 fecal samples/dog, with each fecal sample collected on 2 consecutive days with 28 days between sample collection periods (ie, days 1 and 2 followed by days 28 and 29). Reference range values for fecal IgA concentration were 0.22 to 3.24 mg/g of feces.

Conclusions and Clinical Relevance— Methods of fecal IgA extraction and quantification used in our study allow for identification of dogs with consistently low fecal IgA concentrations. Use of these techniques will enable future investigations into possible associations between low fecal IgA concentrations and signs of gastrointestinal disease in dogs.

Abstract

Objective—To develop a fecal sample collection strategy and quantification method for measurement of fecal IgA concentrations in dogs.

Sample Population—Fecal samples from 23 healthy pet dogs of various breeds.

Procedures——Immunoglobulin A was extracted from fecal samples. An ELISA for the measurement of fecal IgA concentrations was established and analytically validated. Intraindividual variation of fecal IgA was determined by calculation of coefficients of variation. A sample collection strategy was developed on the basis of results of intraindividual variation of fecal IgA concentrations. A reference range for fecal IgA concentrations was determined.

Results—The method for extraction and quantification of fecal IgA was determined to be sufficiently sensitive, reproducible, accurate, and precise. On the basis of the intraindividual variability of our results, the determined fecal sample collection strategy required analysis of a total of 4 fecal samples/dog, with each fecal sample collected on 2 consecutive days with 28 days between sample collection periods (ie, days 1 and 2 followed by days 28 and 29). Reference range values for fecal IgA concentration were 0.22 to 3.24 mg/g of feces.

Conclusions and Clinical Relevance— Methods of fecal IgA extraction and quantification used in our study allow for identification of dogs with consistently low fecal IgA concentrations. Use of these techniques will enable future investigations into possible associations between low fecal IgA concentrations and signs of gastrointestinal disease in dogs.

Contributor Notes

Supported by a grant from Masterfoods, Vernon, Calif.

Presented in part at the 2004 American College of Veterinary Internal Medicine Forum, Minneapolis, May 2004.

Address correspondence to Dr. Tress.