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 [email protected]
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