High-Resolution Melting Curve Analysis of the 16S Ribosomal Gene to Detect and Identify Pathogenic and Saprophytic Leptospira Species in Colombian Isolates

The American Journal of Tropical Medicine and Hygiene
Ronald Guillermo Peláez SanchezPiedad Agudelo-Flórez

Abstract

AbstractIt is important to identify the circulating Leptospira agent to enhance the performance of serodiagnostic tests by incorporating specific antigens of native species, develop vaccines that take into account the species/serovars circulating in different regions, and optimize prevention and control strategies. The objectives of this study were to develop a polymerase chain reaction (PCR)-high-resolution melting (HRM) assay for differentiating between species of the genus Leptospira and to verify its usefulness in identifying unknown samples to species level. A set of primers from the initial region of the 16S ribosomal gene was designed to detect and differentiate the 22 species of Leptospira. Eleven reference strains were used as controls to establish the reference species and differential melting curves. Twenty-five Colombian Leptospira isolates were studied to evaluate the usefulness of the PCR-HRM assay in identifying unknown samples to species level. This identification was confirmed by sequencing and phylogenetic analysis of the 16S ribosomal gene. Eleven Leptospira species were successfully identified, except for Leptospira meyeri/Leptospira yanagawae because the sequences were 100% identical. The 25 isolates from h...Continue Reading

References

Aug 1, 1994·Molecular Biotechnology·J A WagenaarB A Van der Zeijst
Apr 9, 2001·Clinical Microbiology Reviews·P N Levett
Jul 9, 2002·BMC Infectious Diseases·Lee D SmytheDavid B McKay
May 27, 2003·Clinical Chemistry·Carl T WittwerRobert J Pryor
Dec 4, 2003·The Lancet Infectious Diseases·Ajay R BhartiUNKNOWN Peru-United States Leptospirosis Consortium
Feb 1, 2005·Molecular and Cellular Probes·Raghavan U M PalaniappanBruce Roe
Jul 12, 2005·FEMS Microbiology Letters·Fabrice MerienGuy Baranton
Sep 9, 2005·Current Opinion in Infectious Diseases·Alan J A McBrideAlbert I Ko
Mar 4, 2006·International Journal of Systematic and Evolutionary Microbiology·Paul N LevettArnold G Steigerwalt
Oct 6, 2006·FEMS Microbiology Letters·Raven E Reitstetter
Sep 12, 2007·Bioinformatics·M A LarkinD G Higgins
Apr 18, 2008·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Ma de los A ValverdeRudy A Hartskeerl
Dec 17, 2008·Diagnostic Microbiology and Infectious Disease·Xu'ai LinJie Yan
Apr 7, 2009·Veterinary Microbiology·Ben Adler, Alejandro de la Peña Moctezuma
May 7, 2009·Human Mutation·Rolf H A M VossenJohan T den Dunnen
May 30, 2009·Human Mutation·Carl T Wittwer
Jun 18, 2010·The Veterinary Journal·Barbara SzonyiAlbert I Ko
Sep 22, 2010·PLoS Neglected Tropical Diseases·Renee L Galloway, Paul N Levett
Sep 18, 2012·International Journal of Systematic and Evolutionary Microbiology·L SmytheUNKNOWN International Committee on Systematics of Prokaryotes Subcommittee on the Taxonomy of Leptospiraceae
Nov 15, 2012·The American Journal of Tropical Medicine and Hygiene·Claudia M E Romero-VivasAndrew K I Falconar
Jan 23, 2013·Revista peruana de medicina experimental y salud pública·Paola RiveraManuel Céspedes
Jan 30, 2013·PLoS Neglected Tropical Diseases·Siriphan BoonsilpSharon J Peacock
Mar 22, 2013·PLoS Neglected Tropical Diseases·Pascale BourhyMathieu Picardeau
Sep 25, 2014·International Journal of Systematic and Evolutionary Microbiology·Pascale BourhyMathieu Picardeau
Sep 18, 2015·PLoS Neglected Tropical Diseases·Federico CostaAlbert I Ko

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Citations

May 17, 2018·Frontiers in Microbiology·Roman ThibeauxCyrille Goarant
Jun 19, 2021·Science Progress·Wannapimol KriangwanichChatchote Thitaram

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