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 ORIGINAL ARTICLE
Year : 2002  |  Volume : 20  |  Issue : 1  |  Page : 25-28

Arbitrarily primed PCR- A rapid and simple method for typing of leptospiral serovars


1 Department of Animal Biotechnology, Madras Veterinary College, Chennai - 600 007, India
2 Lister Laboratory, Chennai 600 034, India

Correspondence Address:
P Ramadass
Department of Animal Biotechnology, Madras Veterinary College, Chennai - 600 007
India
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Source of Support: None, Conflict of Interest: None


PMID: 17657019

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PURPOSE: To investigate the use of arbitrarily primed polymerase chain reaction (AP-PCR) for typing of leptospiral serovars. METHODS: AP-PCR was adopted for identification of laboratory strains of leptospires and leptospiral cultures at serovar level. A primer of 12 bp was used for amplifying DNA of 13 laboratory strains of leptospires as well as culture pellets of leptospires. RESULTS: Each serovar produced distinct DNA fingerprint which was characteristic for each serovar. These patterns were used for typing of 81 serum culture samples obtained from human leptospiral cases. Of these samples, 39 could be typed based on AP-PCR fingerprints belonging to serovars autumnalis, pomona, canicola, javanica, icterohaemorrhagiae, patoc and pyrogenes. These results were confirmed by RAPD fingerprinting of the DNA samples of the respective leptospiral serovars after culturing -*them in EMJH media. One of the important findings of this work was that straight culture sample could be used for AP-PCR assay, without purification of DNA. By having more number of AP-PCR reference fingerprints, more serovars could be typed. CONCLUSIONS: AP-PCR technique provides great potential for simple and rapid identification of leptospires at serovar level, which could be useful in molecular epidemiological studies of leptospirosis.






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2004 - Indian Journal of Medical Microbiology
Published by Wolters Kluwer - Medknow

Online since April 2001, new site since 1st August '04