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Journal of Clinical Microbiology, 10 1997, 2612-2617, Vol 35, No. 10
Copyright © 1997 by the American Society for Microbiology. All rights reserved.

Differentiation of Leptospira interrogans isolates by IS1500 hybridization and PCR assays

RL Zuerner and CA Bolin
Leptospirosis Reference Center, National Animal Disease Center, USDA Agriculture Research Service, Ames, Iowa 50010, USA. rzuerner@nadc.ars.usda.gov

Genetic variability among Leptospira interrogans (sensu stricto) serovars was assessed by Southern blot hybridization and PCR analyses. The experiments used probes directed to sequences in a recently described insertion element, IS1500. Hybridization analysis showed that IS1500 was present on polymorphic fragments and that differences in these patterns could be used to identify serovars. Hybridization analysis was also useful in discriminating between serovar pomona type kennewicki isolates, making possible the identification of 15 previously unrecognized genetic groups. A PCR assay was developed in which the primers are positioned near the terminal inverted repeats of the element and directed outward. This assay yielded characteristic amplification patterns from isolates, allowing them to be identified. We applied these assays to several new animal isolates of L. interrogans from Nicaragua, which recently had an outbreak of human leptospirosis. Three groups of isolates were identified: one strain of serovar pomona type kennewicki and two genetically distinct groups of isolates which may be genetic intermediates between serovars canicola and portlandvere. The IS-based typing assays described should be useful for epidemiological analysis of leptospirosis.

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