[MARMAM] New publication: Development of a real-time PCR for the detection of pathogenic Leptospira spp. in California sea lions.

[David Wu - NOAA Affiliate]

Dear colleagues,

We are pleased to announce that the publication of our article is now
available online:

Qingzhong Wu, Katherine C. Prager, Tracey Goldstein, David P. Alt, Renee L.
Galloway, Richard L. Zuerner, James O. Lloyd-Smith and Lori Schwacke.
2014. Development
of a real-time PCR for the detection of pathogenic  *Leptospira* spp.
in California
sea lions. Diseases of Aquatic Organisms. DOI: 10.3354/dao02752

http://www.int-res.com/abstracts/dao/v110/n3/p165-172/



Abstract:

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.
Comparisons
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
tissue samples.



Best regards,

Qingzhong (David) Wu

Molecular microbiologist

-- 

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
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