Evaluation of the use of selective PCR amplification of LPS biosynthesis genes for molecular typing of leptospira at the serovar level.
Leptospirosis is an important epidemic zoonosis worldwide. Currently, there are more than 250 Leptospira pathogenic serovars known that can potentially infect humans. Conventional classification of leptospires with the serovar as the basic taxon, based on serological recognition of lipopolysaccharide (LPS) composition does not correlate well with species determination, based on general genomic features. Here, we investigate the selective amplification of polymorphic regions from the LPS biosynthesis loci (rfb) as a potential tool for serovar typing of Leptospira interrogans species. Eight pairs of primers were designed to target six ORFs from the rfb operon with varying levels of sequence polymorphism. They were tested both separately and multiplexed. Half of these primer pairs produced serovar-specific amplicons, allowing the identification of some specific serovars and also groups of serovars. It was shown that the serovar classification of Leptospira can be accessed by selective amplification of rfb operons in some cases, which may permit a parallel between the serological and the genomic classifications of Leptospira. As a conclusion, the selective amplification of rfb generated promising and already useful results, but it appears necessary to characterize a larger variety of Leptospira genomes or rfb operons to fully develop this method.
Bezerra da Silva
Josefa
J
Centro de Biotecnologia, Instituto Butantan, 05503-900, São Paulo, SP, Brazil.
Carvalho
Eneas
E
Hartskeerl
Rudy A
RA
Ho
Paulo L
PL
D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D005838:Genotype; D006801:Humans; D007921:Leptospira interrogans / Q000145:classification* / Q000235:genetics*; D007922:Leptospirosis / Q000382:microbiology; D008070:Lipopolysaccharides / Q000235:genetics*; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic
D017931:DNA Primers; D008070:Lipopolysaccharides