J Vet Sci.  2005 Sep;6(3):231-237.

Simultaneous detection of Lawsonia intracellularis, Brachyspira hyodysenteriae and Salmonella spp. in swine intestinal specimens by multiplex polymerase chain reaction

• Suh DK
• Song JC

Abstract

A multiplex PCR assay was developed for the simultaneous detection of the etiologic agents associated with porcine proliferative enteropathies (PPE), swine dysentery (SD)and porcine salmonellosis (PS)in a single reaction using DNA from swine intestinal samples. Single and multiplex PCR amplification of DNA from Lawsonia intracellularis, Salmonella typhimurium and Brachyspira hyodysenteriae with each primer set produced fragments of the predicted size without any nonspecific amplification, 210-bp, 298-bp and 403-bp bands, respectively. The single PCR assay could detect as little as 100 pg of purified DNA of S. typhimurium and L. intracellularis, and 50 pg of B.hyodysenteriae, respectively. However, multiplex PCR turned out to be 10 times lower sensitivity with S. typhimurium compared with single PCR. With 23 swine intestinal specimens suspected of having PPE, SD and/or PS, the multiplex PCR assay showed identical results with conventional methods except one. In conclusion, this multiplex PCR is a feasible alternative to standard diagnostic methods for detection of L. intracellularis, B. hyodysenteriae and Salmonella spp. from swine intestinal specimens.

Keyword

Brachyspira hyodysenteriae; Lawsonia intracellularis; Multiplex PCR; Salmonella spp

MeSH Terms

Animals
Desulfovibrionaceae Infections/microbiology/veterinary
Intestines/microbiology
Lawsonia Bacteria/*isolation&purification
Polymerase Chain Reaction/*methods/veterinary
Salmonella/*isolation&purification
Salmonella Infections, Animal/diagnosis
Sensitivity and Specificity
Spirochaetales/*isolation&purification
Spirochaetales Infections/microbiology/veterinary
Swine
Swine Diseases/*diagnosis/*microbiology