[MARMAM] New publication: Development of a real-time PCR for the detection of pathogenic Leptospira spp. in California sea lions.
David Wu - NOAA Affiliate
david.wu at noaa.gov
Mon Aug 11 07:26:15 PDT 2014
We are pleased to announce that the publication of our article is now
Qingzhong Wu, Katherine C. Prager, Tracey Goldstein, David P. Alt, Renee L.
Galloway, Richard L. Zuerner, James O. Lloyd-Smith and Lori Schwacke.
of a real-time PCR for the detection of pathogenic *Leptospira* spp.
sea lions. Diseases of Aquatic Organisms. DOI: 10.3354/dao02752
Several real-time PCR assays are currently used for detection of pathogenic
*Leptospira* spp.; however, few methods have been described for the
successful evaluation of clinical urine samples. This study reports a
rapid assay for the detection of pathogenic *Leptospira *spp. in California
sea lions (*Zalophus californianus*) using real-time PCR with primers and a
probe targeting the *lipL32* gene. The PCR assay had high analytic
sensitivity - the limit of detection was three genome copies per PCR volume
using *L.** interrogans* serovar Pomona DNA; and 100% analytic specificity
- it detected all pathogenic leptospiral serovars tested and none of the
non-pathogenic *Leptospira* species *L. biflexa *and* L. meyeri* serovar
Semaranga*,* the intermediate species *L. inadai*, or the non-*Leptospira*
pathogens tested. Our assay had an amplification efficiency of 1.00.
between the real-time PCR assay and culture isolation for detection of
pathogenic *Leptospira *spp*. *in urine and kidney tissue samples from
California sea lions showed that samples were more often positive by
real-time PCR than by culture methods. Inclusion of an internal
amplification control in the real-time PCR assay showed no inhibitory
effects in PCR negative samples. These studies indicated that our
real-time PCR assay has high analytic sensitivity and specificity for the
rapid detection of pathogenic *Leptospira *species in urine and kidney
Qingzhong (David) Wu
Qingzhong (David) Wu, JHT, Inc. - Contractor
Hollings Marine Laboratory
National Ocean Service
331 Ft. Johnson Rd.
Charleston, SC 29412
Office 843-762-8940david.wu at noaa.gov
-------------- next part --------------
An HTML attachment was scrubbed...
More information about the MARMAM