A LightCycler real-time PCR hybridization probe-based assay that detects a conserved region of the16S rRNA gene of pathogenic but not saprophytic @entity2928 species was developed for the rapid detection of pathogenic leptospires directly from processed tissue samples. In addition, a differential PCR specific for saprophytic leptospires and a control PCR targeting the porcine @entity28249 gene were developed. To assess the suitability of these PCR methods for diagnosis, a trial was performed on kidneys taken from adult @entity1707 with evidence of @entity281 , primarily a history of reproductive disease and serological evidence of exposure to pathogenic leptospires (n=180) and aborted @entity1707 foetuses (n=24). Leptospire DNA was detected by the 'pathogenic' specific PCR in 25 tissues (14%) and the control @entity28249 PCR was positive in all 204 samples confirming DNA was extracted from all samples. No leptospires were isolated from these samples by culture and no positives were detected with the 'saprophytic' PCR. In a subsidiary experiment, the 'pathogenic' PCR was used to analyse kidney samples from rodents (n=7) collected as part of vermin control in a zoo, with show animals with high microagglutination titres to @entity2928 species, and five were positive. Fifteen PCR amplicons from 1 @entity19 , 2 @entity35 and 14 @entity1707 kidney samples, were selected at random from positive PCRs (n=30) and sequenced. Sequence data indicated @entity2928 DNA in the @entity1707 and @entity35 samples and L. inadai DNA, which is considered of intermediate pathogenicity, in the @entity19 sample. The only successful culture was from this @entity19 kidney and the isolate was confirmed to be L. inadai by classical serology. These data suggest this suite of PCRs is suitable for testing for the presence of pathogenic leptospires in @entity1707 herds where @entity5852 and @entity249 occur and potentially in other animals such as rodents.

The development of a real-time PCR to detect pathogenic XXXX species in kidney tissue.

@entity1707 :: ('9823', 'Species') :: ['pigs', 'pig'] @entity249 :: ('MESH:D007246', 'Disease') :: ['infertility'] @entity5852 :: ('MESH:D000031', 'Disease') :: ['abortions'] @entity28249 :: ('396797(Tax:9823)', 'Gene') :: ['beta-actin'] @entity19 :: ('10090', 'Species') :: ['mouse'] @entity2928 :: ('173', 'Species') :: ['Leptospira', 'L. interrogans'] @entity281 :: ('MESH:D007239', 'Disease') :: ['leptospiral infection'] @entity35 :: ('10116', 'Species') :: ['rat']
@entity2928 :: (173,Species) :: ['Leptospira']
