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Journal of Clinical Microbiology, March 2002, p. 984-987, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.984-987.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Application of a 5' Nuclease Assay for Detection of Lawsonia intracellularis in Fecal Samples from Pigs

R. H. Lindecrona,* T. K. Jensen, P. H. Andersen, and K. Møller

Danish Veterinary Laboratory, DK-1790 Copenhagen V, Denmark

Received 13 June 2001/ Returned for modification 23 September 2001/ Accepted 30 December 2001

A 5' nuclease assay was developed to detect Lawsonia intracellularis in porcine fecal samples. The specific probe and primers were chosen by using the 16S ribosomal DNA gene as a target. The 5' nuclease assay was used with a total of 204 clinical samples, and the results were compared to those of immunohistochemistry (IM) on ileal sections of the same animals. There was 91% agreement between the results of IM and the 5' nuclease assay. In the 5' nuclease assay, 111 (54%) of the pigs tested positive for L. intracellularis infection, with a mean cycle threshold (Ct) value of 27.2, whereas 98 (48%) of the pigs tested positive by IM. On average, the Ct and {Delta}Rn values for the positive samples were 27.2 (standard deviation [SD], 3.7) and 1.6 (SD, 0.7), respectively. A Ct value of 27.2 corresponds to a fecal excretion of approximately 107 L. intracellularis cells per g of feces. Furthermore, a total of 40 fecal samples derived from a herd known to be free from infection with L. intracellularis all tested negative, with a Ct value of 40. By using a Ct value of 36 as the cutoff limit, the detection limit of the assay was 1 L. intracellularis cell per PCR tube. In conclusion, the 5' nuclease assay that has been developed represents an applicable fast method for detection of L. intracellularis in fecal samples, with a sensitivity and specificity comparable to those of IM.

* Corresponding author. Present address: Danish Veterinary and Food Administration, Institute of Food Safety and Toxicology, Mørkhøj Bygade 19, DK-2860 Søborg, Denmark. Phone: 45 33956000. Fax: 45 33956001. E-mail: rhl{at}fdir.dk.

Journal of Clinical Microbiology, March 2002, p. 984-987, Vol. 40, No. 3
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.3.984-987.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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