Identification of new repetitive element in Leptospira interrogans serovar copenhageni and its application to PCR-based differentiation of Leptospira serogroups.

Abstract

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2014-05-02T14:12:01Z No. of bitstreams: 1 Barocchi MA Identification of New....pdf: 2689916 bytes, checksum: b29266c121b560e1b5c6260b77f5c08f (MD5)Made available in DSpace on 2014-05-02T14:12:01Z (GMT). No. of bitstreams: 1 Barocchi MA Identification of New....pdf: 2689916 bytes, checksum: b29266c121b560e1b5c6260b77f5c08f (MD5) Previous issue date: 2001Division of Infectious Diseases and Immunity. School of Public Health. University of California. Berkeley, CaliforniaFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Division of International Medicine and Infectious Diseases. Weil Medical College of Cornell University. New York, New YorkFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, BrasilDivision of Infectious Diseases and Immunity. School of Public Health. University of California. Berkeley, CaliforniaA new repetitive DNA element was identified in an isolate of Leptospira interrogans serovar copenhageni from a patient in Salvador, Brazil. A Sau3A genomic library from this strain was constructed and screened for repetitive DNA elements. An insert of 438 bp (Rep1) from one library clone hybridized to multiple chromosomal DNA fragments resolved electrophoretically after digestion with BamHI, HindIII, and MfeI. A single oligonucleotide primer, designated iRepl, was designed to generate multiple PCR amplicons of various electrophoretic mobilities in a PCR typing method. The method distinguished strains belonging to the eight pathogenic and three saprophytic species of the genus Leptospira. Clinical isolates obtained during urban epidemics between 1996 and 1998 in Salvador, Brazil, were analyzed by this PCR method. Although the iRep1 primer was unable to discriminate strains among L. interrogans serovar copenhageni isolates, it was able to differentiate strains belonging to different species and serogroups of Leptospira identified in Salvador. This PCR-based method may provide a faster and less expensive alternative to serologic tests used in reference laboratories