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Clinical and Diagnostic Laboratory Immunology, June 2005, p. 693-699, Vol. 12, No. 6
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.6.693-699.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

J. J. Kroll,1* M. A. Eichmeyer,1 M. L. Schaeffer,1 S. McOrist,2 D. L. Harris,3 and M. B. Roof1

Department of Research and Development, Boehringer Ingelheim Vetmedica Inc., 2501 North Loop Drive, Ames, Iowa 50010,1 QAF Industries, Corowa, NSW 2646, Australia,2 Iowa State University, Ames, Iowa 500113

Received 20 December 2004/ Returned for modification 7 March 2005/ Accepted 12 April 2005

An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specificity and sensitivity values based on 80 known positive and 80 known negative serum samples from controlled experimental trials were 93.7% and 88.7%, respectively. Serological results from a controlled L. intracellularis challenge exposure study confirmed the high specificity and sensitivity of this assay (100% and 99.5%, respectively). Comparisons between the LPS ELISA and the IFAT in detecting anti-Lawsonia antibodies in this controlled study revealed significantly more LPS ELISA-positive pigs than IFAT-positive pigs on days 21, 28, 35, and 42 (P = 0.003, 0.030, 0.002, and 0.006, respectively). This indirect ELISA (LPS ELISA) test is an improved method of detecting antibodies in pigs soon after exposure to L. intracellularis, regardless of isolate type (vaccine or wild type) in experimental studies. The LPS ELISA may be used as a tool to support future research trials on vaccine efficacy and to further understand the immune response induced by L. intracellularis.

* Corresponding author. Mailing address: Department of Research and Development, Boehringer Ingelheim Vetmedica Inc., 2501 North Loop Drive, Ames, Iowa 50010. Phone: (515) 296-6625. Fax: (515) 296-7937. E-mail: jkroll{at}bi-vetmedica.com.

Clinical and Diagnostic Laboratory Immunology, June 2005, p. 693-699, Vol. 12, No. 6
1071-412X/05/$08.00+0     doi:10.1128/CDLI.12.6.693-699.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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