Assessment of Southern blot ribotyping for differentiation of Leptospira strains isolated from field rats.
A Southern blot ribotyping based on EcoRV and HindIII digestion with two 16S and 23S rDNA probes for differentiating 27 Leptospira serovars was developed. The results between ribotyping and serotyping among 40 leptospiral strains isolated from field rats trapped in the northeastern region of Thailand during 1999-2000, were compared. A combination of Southern blot ribotyping, using EcoRV or HindIII digestion with both 16S and 23S rDNA as the probes, successfully typed 27 Leptospira serovars into 24 ribotypes with the discriminatory index (D) values of 0.99. The 16S- and 23S-EcoRV ribopatterns produced 17 and 9 profiles, respectively, with D values of 0.95 and 0.63, respectively. Ribopatterns of HindIII from both specific probes yielded 17 patterns. The D values of 16S- and 23S-HindIII ribopatterns were 0.94 and 0.93, respectively. With EcoRV digestion, the 16S rDNA probe was more discriminative than the 23S rDNA probe for differentiating Leptospira serovars. Moreover, the 16S-EcoRV (11 profiles), 16S-HindIII (11 profiles), and 23S-HindIII (10 profiles) ribopatterns produced higher numbers of distinct and unique profiles than the 23S-EcoRV (5 profiles). The results showed 100% concordance between ribotyping and serotyping, leading to all 40 isolates being successfully typed. The current study revealed that ribotyping as a quick and powerful tool for differentiating Leptospira serovars, has potential value in epidemiological studies.
Kositanont
Uraiwan
U
Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. gruks@mahidol.ac.th
Chotinantakul
Kamonnaree
K
Phulsuksombati
Duangporn
D
Tribuddharat
Chanwit
C
D000818:Animals; D015139:Blotting, Southern / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D007919:Leptospira / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D007922:Leptospirosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000737:chemistry / Q000235:genetics; D051381:Rats; D021521:Ribotyping / Q000379:methods*; D012703:Serotyping; D013785:Thailand
D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S; D004262:DNA Restriction Enzymes