Polymerase chain reaction assay specific for pathogenic Leptospira based on the gene hap1 encoding the hemolysis-associated protein-1.
In this study, we used Southern hybridization of genomic DNA with the integral hap1 gene as a probe to show that this gene is only present in pathogenic Leptospira strains. We then selected PCR primers based on the hap1 gene, and tested them on several Leptospira strains and biological samples. Specific amplification was obtained for all pathogenic strains tested. Negative PCR results were observed with all saprophytic leptospire strains used as well as with other spirochetes and bacteria commonly found in biological samples. The results of direct PCR performed on biological samples, such as blood, urine or kidneys correlated with the results obtained with the classical Leptospira tests (culture and MAT). A PCR assay based on this gene would be a very useful tool for the rapid, sensitive and specific identification of pathogenic leptospires in samples for diagnosis or epidemiological survey.
Branger
Christine
C
Leptospira Medical and Molecular Bacteriology Unit, ENVN, BP 40706, 44307 Nantes Cedex 03, France.
Blanchard
Béatrice
B
Fillonneau
Catherine
C
Suard
Isabelle
I
Aviat
Florence
F
Chevallier
Bruno
B
André-Fontaine
Geneviève
G
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers; D004283:Dog Diseases / Q000175:diagnosis* / Q000382:microbiology; D004285:Dogs; D006460:Hemolysin Proteins; D006801:Humans; D007919:Leptospira / Q000145:classification* / Q000235:genetics / Q000472:pathogenicity*; D007922:Leptospirosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers; D006460:Hemolysin Proteins; C439630:hemolysis-associated protein 1