Protocol

PCR Detection of Microbial Pathogens

Volume 943 of the series Methods in Molecular Biology pp 257-266

Date:

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

  • Robyn Anne StoddardAffiliated withNational Center for Zoonotic, Vector-Borne, and Enteric Diseases, Centers for Disease Control and Prevention Email author 

* Final gross prices may vary according to local VAT.

Get Access

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