JVDI Advertisement
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Journal of Veterinary Diagnostic Investigation Vol. 22 Issue 1, 70-73
Copyright © 2010 by the American Association of Veterinary Laboratory Diagnosticians
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Google Scholar
Right arrow Articles by Richter, B.
Right arrow Articles by Weissenböck, H.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Richter, B.
Right arrow Articles by Weissenböck, H.

Brief Research Reports

A TaqMan quantitative polymerase chain reaction assay for the detection of Lawsonia intracellularis in fecal and tissue samples from pigs

Barbara Richter1, Andrea Ladinig, Nora Nedorost and Herbert Weissenböck

Correspondence: 1Corresponding Author: Barbara Richter, Institute of Pathology and Forensic Veterinary Medicine, University of Veterinary Medicine Vienna, Veterinaerplatz 1, 1210 Vienna, Austria, e-mail: Barbara.Richter{at}vetmeduni.ac.at

In the present study, a TaqMan quantitative polymerase chain reaction (qPCR) assay for detecting the 16S ribosomal RNA gene of Lawsonia intracellularis in porcine native ileal mucosal scrapings, fecal samples, and formalin-fixed and paraffin-embedded (FFPE) ileal samples is described. Samples from 62 pigs were examined. The results of the qPCR were compared with results obtained with conventional detection methods (PCR, immunohistochemistry, in situ hybridization, and silver staining) from a previous study and correlated well. The qPCR assay proved to be very sensitive and specific. In particular, the sensitivity of TaqMan PCR was significantly higher than conventional PCR on FFPE tissues because of a much shorter amplicon. A higher number of copies per gram of sample material was detected in native mucosa and FFPE tissue compared with feces, especially in highly positive animals. The detection limit for the qPCR was at 4 copies per well in native mucosal scrapings and 18 copies per well in feces and FFPE tissue, respectively. Inhibition of the qPCR reaction was checked by simultaneous detection of a recombinant beta-actin plasmid using a second fluorescent probe. A decreased signal from this internal control plasmid revealed inhibition of the PCR reaction in 21% of native mucosal samples and 1.6% of fecal samples. With a 10-fold dilution of template, the inhibition could be overcome.

Key Words: Lawsonia intracellularis • pigs • porcine proliferative enteropathy • quantitative polymerase chain reaction • real-time polymerase chain reaction • TaqMan






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2010 by the American Association of Veterinary Laboratory Diagnosticians, Inc.