Application of pulsed-field gel electrophoresis for the discrimination of leptospiral isolates in Brazil.
OBJECTIVE
Leptospirosis is a public health problem worldwide. Traditionally, microscopic agglutination test (MAT) and cross-agglutinin absorption test (CAAT) are used to identify leptospires. However, these techniques are laborious and time-consuming, requiring the maintenance of a collection of more than 200 reference strains and correspondent rabbit antisera. The purpose of this study was to evaluate the pulsed-field gel electrophoresis (PFGE) method for discrimination of Leptospira serovars.


RESULTS
Fourteen clinical isolates of Leptospira spp. were analysed by MAT before being characterized by PFGE. The isolates were compared with a library of 206 different reference Leptospira serovars. All the isolates gave clear profiles with high resolution. PFGE and MAT results were in agreement for all clinical isolates evaluated. Twelve isolates were classified as serovar Icterohaemorrhagiae/Copenhageni by PFGE. By MAT, these isolates were classified as serogroup Icterohaemorrhagiae with titres ranging from 3200 to 25 600. Two isolates were classified as serovar Canicola by PFGE, and as serogroup Canicola by MAT with titres higher than 3200.


CONCLUSIONS
PFGE offers the advantages of simple, reliable and reproducible results.


CONCLUSIONS
PFGE provides a convenient tool for the identification of clinical isolates.
Department of Medical Biology, Bacteriology, Instituto Adolfo Lutz, Sao Paulo, Brazil. eliete_romero @yahoo.com.br
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Romero
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Blanco
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Galloway
D015373:Bacterial Typing Techniques; D001938:Brazil; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007922:Leptospirosis / Q000382:microbiology*; D010802:Phylogeny