HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Levett, P. N Articles by Mayer, L. W Search for Related Content PubMed PubMed Citation Articles by Levett, P. N Articles by Mayer, L. W Agricola Articles by Levett, P. N Articles by Mayer, L. W

Detection of pathogenic leptospires by real-time quantitative PCR

Paul N Levett, Roger E Morey, Renee L Galloway, Danielle E Turner, Arnold G Steigerwalt and Leonard W Mayer

Meningitis and Special Pathogens Branch, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA, USA

Correspondence Paul N. Levett plevett{at}health.gov.sk.ca

Received August 9, 2004
Accepted September 30, 2004

Definitive diagnosis of leptospirosis has traditionally depended upon the isolation of leptospires from clinical specimens or the demonstration of seroconversion in paired acute and convalescent serum samples. Both of these approaches require expertise not routinely available in clinical laboratories and usually result in delayed diagnosis. Conventional PCR assays have been developed, but all have limitations which have restricted their widespread use. In order to overcome these limitations, a real-time PCR assay was developed using a 423 bp target on the lipL32 gene, which is conserved among pathogenic serovars of Leptospira. Reactions were monitored by SYBR green fluorescence and melting curve analysis. Representative serovars from 16 species of Leptospira and over 40 species of other bacteria and fungi were tested. Positive results were obtained with all pathogenic leptospiral serovars, with the exception of Leptospira fainei serovar Hurstbridge. The analytical sensitivity of this assay was 3 genome equivalents per reaction; approximately 10 genome equivalents were detectable in human urine. Leptospiral DNA was amplified from blood containing EDTA or citrate anticoagulants, but heparin, sodium polyanetholesulfonate and saponin were inhibitory. The assay successfully detected leptospiral DNA from serum and urine samples of patients with leptospirosis. This assay has the potential to facilitate rapid, sensitive diagnosis of acute leptospirosis.

This article has been cited by other articles:

				A. Roczek, C. Forster, H. Raschel, S. Hormansdorfer, K.-H. Bogner, A. Hafner-Marx, H. Lepper, G. Dobler, M. Buttner, and A. Sing Severe course of rat bite-associated Weil's disease in a patient diagnosed with a new Leptospira-specific real-time quantitative LUX-PCR J. Med. Microbiol., May 1, 2008; 57(5): 658 - 663. [Abstract] [Full Text] [PDF]

				B. Guerra, T. Schneider, E. Luge, A. Draeger, V. Moos, C. Loddenkemper, A. Jansen, and K. Nockler Detection and characterization of Leptospira interrogans isolates from pet rats belonging to a human immunodeficiency virus-positive patient with leptospirosis J. Med. Microbiol., January 1, 2008; 57(1): 133 - 135. [Full Text] [PDF]

				K. S. A. MYINT, R. V. GIBBONS, C. K. MURRAY, K. RUNGSIMANPHAIBOON, W. SUPORNPUN, R. SITHIPRASASNA, M. R. GRAY, C. PIMGATE, M. P. MAMMEN JR, and D. R. HOSPENTHAL LEPTOSPIROSIS IN KAMPHAENG PHET, THAILAND Am J Trop Med Hyg, January 1, 2007; 76(1): 135 - 138. [Abstract] [Full Text] [PDF]