PCR Detection of Microbial Pathogens pp 257-266

Part of the Methods in Molecular Biology book series (MIMB, volume 943)

Detection of Pathogenic Leptospira spp. Through Real-Time PCR (qPCR) Targeting the LipL32 Gene

Protocol

Abstract

Rapid diagnosis of leptospirosis, through culture and/or serology, can be difficult without proper expertise and is often delayed due to the length of time required to obtain results. Polymerase chain reaction (PCR), more specifically the real-time detection of the amplified PCR product, is a methodology that can provide a diagnosis in a timelier manner compared to culture and serology. There are a limited number of real-time PCR (qPCR) assays for detecting Leptospira and not all of these assays are able to distinguish pathogenic from nonpathogenic species. In addition, there are a variety of probe technologies and qPCR instruments that are utilized with these assays. This chapter presents a qPCR assay that targets lipL32, a gene which is present only in pathogenic Leptospira spp. This assay utilizes a TaqMan probe and instructions for use on either the Lightcycler 1.2 (Roche Diagnostics, Indianapolis, IN) or the ABI 7500 (Applied Biosystems, Foster City, CA) are provided.

Key words

Leptospira Leptospirosis Real-time PCR TaqMan Diagnosis LipL32 

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Centers for Disease Control and PreventionAtlantaUSA

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