Ring test evaluation of the repeatability and reproducibility of a Porcine reproductive and respiratory syndrome virus oral fluid antibody enzyme-linked immunosorbent assay

Apisit Kittawornrat, Chong Wang, Gary Anderson, Andrea Ballagi, André Broes, Suzanne Carman, Kent Doolittle, Judith Galeota, John Johnson, Sergio Lizano, Eric Nelson, Devi Patnayak, Roman Pogranichniy, Anna Rice, Gail Scherba, Jeffrey Zimmerman

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

The precision of a Porcine reproductive and respiratory syndrome virus (PRRSV) oral fluid antibody enzyme-linked immunosorbent assay (ELISA) was evaluated by calculating reliability coefficients for assay repeatability (within laboratory) and assay reproducibility (between laboratories). Randomly ordered oral fluid samples of known (n = 39) and unknown (n = 224) PRRSV antibody status were tested in 12 diagnostic laboratories. Each laboratory tested the samples twice, first using an antibody ELISA kit and reagents provided to them (phase 1) and then using an ELISA kit and reagents configured in their respective laboratory (phase 2). Repeatability (within laboratory) reliability coefficients calculated using results from samples of known PRRSV antibody status ranged from 0.724 to 0.997 in phase 1 and from 0.953 to 0.998 in phase 2. Reproducibility (between laboratories) reliability coefficients were calculated for 3 conditions: case 1-samples of unknown status (n = 224); case 2-samples of known status (n = 39), and case 3-all samples (n = 263). Among the 3 cases, reliability coefficients ranged from 0.937 to 0.964 in phase 1 and from 0.922 to 0.935 in phase 2. For case 3, it was estimated that 96.67% of the total variation in phase 1 and 93.21% in phase 2 could be attributed to the oral fluid samples themselves. Overall, the PRRSV oral fluid antibody ELISA was highly repeatable and reproducible. The current study supports the routine use of this test in laboratories providing diagnostic service to pig producers.

Original languageEnglish (US)
Pages (from-to)1057-1063
Number of pages7
JournalJournal of Veterinary Diagnostic Investigation
Volume24
Issue number6
DOIs
StatePublished - Nov 2012

Bibliographical note

Funding Information:
The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: The study was supported in part by Pork Checkoff funds distributed through the National Pork Board (PO Box 9114, Des Moines, IA 50306) and the PRRS CAP, USDA NIFA award 2008-55620-1932. HerdChek PRRS 3X ELISA kits were provided by IDEXX Laboratories Inc., Westbrook, ME.

Keywords

  • Enzyme-linked immunosorbent assay
  • Porcine reproductive and respiratory syndrome virus
  • oral fluid
  • reliability coefficient
  • repeatability
  • reproducibility

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