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Original Articles
Characterization of Korean Leptospira interrogans Isolates by Pulsed-Field Gel Electrophoresis of Not I Digests of DNA
Eun-Kyoung Yim, M.S.*,, In-Ae Chang, M.S.*,, Jin-Sang Lee, M.S.*,, Yoon-Won Kim, M.D.*,, Soo-Dong Woo, Ph.D., Jee-Yin Ahn, Ph.D., Seung-Kee Park* and Min-Kee Cho*,
Department of Microbiology*, College of Medicine, Hallym University, Chuncheon, Institute of Medical Science, Hallym University, Chuncheon, and Department of Agricultural Biology, College of Agriculture, Chungbuk University, Korea
Vol.36 Num.2 (p59~67)
Background:Many strains of Leptospira interrogans have been isolated in Korea since 1984. Most isolates were identified as serovar lai by serological methods. The pulsed field gel electrophoresis (PFGE) patterns of Korean isolates have not been investigated currently.
Methods:29 reference strains and 29 Korean isolates of Leptospira interrogans were characterized by PFGE. Chromosomes were digested by the Not I restriction enzyme and subsequently PFGE was performed in CHEF-DRII (Bio Rad Lab) with 3 pulse times (30 seconds 13 hours, 60 seconds 13 hours, 120 seconds 14 hours) at 150 V (4.5 V/cm).
Results:12 serogroup reference strains and most 17 serovars reference strains in the serogroup Icterohaemoffhagie showed the unique Not I restriction patterns. Most isolates identified serologically as serovar lai showed the same PFGE patterns as the serovar lai reference strain. The strain HM3 and 18R identified serologically as new serovars yeonchon and hongchon respectively showed the same PFGE patterns as serovar lai. The strain AP31, CH88-19 and NR13 that were different from serovar lai by serological methods showed the PFGE patterns indistinguishable from serovar lai reference strain. The strain HY2 that was identified as serovar lai, and the strain 30R that was different from serovar lai serologically showed the PFGE patterns slightly different from serovar lai reference strain.
Conclusion:The PFGE profile of most Korean isolates Leptospira interrogans serologically identified as serovar lai is identical to the reference strain serovar lai. PFGE analysis thus may be applied to identify serovar of isolates and to investigate the genetic diversity of related serovar.
Keywords : PFGE, Icterohaemorrhagiae, Leptospira interrogans, Not I