title: Detection of pathogenic Leptospira spp. through real-time PCR (qPCR) targeting the LipL32 gene.
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.
authors.affiliation: National Center for Zoonotic, Vector-Borne, and Enteric Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA. RAStoddard@cdc.gov
authors.forename: Robyn Anne
authors.initials: RA
authors.lastname: Stoddard
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000302:isolation & purification; D006801:Humans; D007919:Leptospira / Q000235:genetics / Q000302:isolation & purification*; D007922:Leptospirosis / Q000175:diagnosis*; D008074:Lipoproteins / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D043262:Ribonuclease P / Q000235:genetics; D013048:Specimen Handling / Q000379:methods
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; C406589:LipL32 protein, Leptospira; D008074:Lipoproteins; D043262:Ribonuclease P