Original scientific paper

Identification and Typing of Leptospira spp. by Restriction Fragment Length Polymorphism (RFLP) of 16S Gene rDNA and Pulsed-field Gel Electrophoresis (PFGE)

Nenad Turk ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Zoran Milas ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Josipa Habuš ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Zrinka Štritof ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Vesna Mojčec ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Zvonko Modrić ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Vilim Starešina ; Veterinarski fakultet Sveučilišta u Zagrebu, Laboratorij za leptospire, Zagreb, Hrvatska
Daniele Postic ; Laboratorie des spirochetes, Institut Pasteur, Paris, France

Abstract

Atotal of 13 Leptospira spp. strains isolated from domestic and wild mammals in different regions of inland Croatia were identified and characterized serologically by microscopic agglutination (MAT) and by modern molecular biology methods; polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) and pulsed field gel electrophoresis (PFGE). After the first typing according to serogroup affinities the isolates appeared to belong to serogroups Pomona (five isolates), Bataviae (two isolates), Canicola (two isolates), Icterohaemorrhagiae (two isolates), Australis (one isolate) and Grippotyphosa (one isolate). RFLP were examined in PCR products from first 330 bp long portion of rrs (16S rDNA gene) digested with endonucleases MnlI and DdeI revealed that all strains belonged to Leptospira interrogans sensu stricto genomic species. Intact genomic DNA from 13 leptospiral isolates was digested with NotI and SgrAI restriction enzymes and analyzed by PFGE. The results showed eight different serovars: Copenhageni (1), Icterohaemorrhagiae (1), Bataviae (2), Pomona (4), Pomona Kennewicki (1), Canicola (2), Lora (1) and Grippotyphosa (1). By revealing the degress of relatedness among 13 leptospiral restriction fingerprints in Croatia we demonstrated the existence of great genetic diversity within one genomic species, Leptospira interrogans sensu stricto especially in the Sava and Drava river valleys.

Keywords

leptospirosis; leptospira; molecular typing; RFLP; PFGE

Hrčak ID:

43725

URI

https://hrcak.srce.hr/43725

Publication date:

2.12.2008.

Article data in other languages: croatian

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