Summary auto-generated
This study describes the first genetic characterization of the outer surface protein C (ospC) gene in Lyme disease spirochetes (Borrelia burgdorferi sensu lato) isolated from rodents in Taiwan. Researchers analyzed seven Taiwan isolates (TWKM1-7) using PCR-RFLP analysis and DNA sequencing. All Taiwan isolates were amplified to produce approximately 630 base pair ospC gene fragments. Using restriction enzyme DraI digestion, all Taiwan isolates displayed identical RFLP pattern type AI, consistent with B. burgdorferi sensu stricto, distinguishing them from B. garinii and B. afzelii genospecies. Sequence analysis revealed the Taiwan isolates were highly homogeneous, sharing 99.3-100% nucleotide sequence identity with each other and with reference B. burgdorferi sensu stricto strains B31, JD1, TB, and CT27985. Phylogenetic analysis confirmed Taiwan isolates clustered closely together with less than 0.5% sequence divergence from these reference strains. The study demonstrates that ospC gene analysis via RFLP patterns and sequence similarity effectively identifies Borrelia genospecies both at inter-species and intra-species levels. Results confirm all Taiwan isolates belong to B. burgdorferi sensu stricto and provide molecular tools for characterizing Lyme disease spirochetes in Taiwan.
Key findings
- All seven Taiwan isolates belong to B. burgdorferi sensu stricto genospecies, confirmed by identical ospC RFLP pattern type AI and high sequence homogeneity (99.3-100%)
- Taiwan isolates demonstrate minimal genetic diversity, with less than 0.5% sequence divergence from reference B. burgdorferi sensu stricto strains B31, JD1, TB, and CT27985
- PCR-RFLP analysis of ospC genes effectively distinguishes between B. burgdorferi genospecies (B. burgdorferi sensu stricto, B. garinii, and B. afzelii) using DraI restriction enzyme digestion patterns
- ospC gene analysis is useful for molecular typing and identification of Borrelia isolates from diverse geographical and biological sources
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Abstract
The outer surface protein C gene (ospC) of Lyme disease spirochaetes (Borrelia burgdorferi sensu lato) was analysed for the first time in Taiwan. The genetic identities of these Taiwan isolates (TWKM17) were determined by restriction fragment length polymorphism (RFLP) analysis and sequence similarities of the PCR-amplified ospC gene amplicons. After cleavage by nuclease DraI, differential fragment patterns of PCR-amplified ospC DNA in relation to different genospecies of Lyme disease spirochaetes were observed and all of these Taiwan isolates were genetically affiliated to the genospecies of B. burgdorferi sensu stricto. The phylogenetic analysis on the sequence similarity of these Taiwan isolates revealed a highly homogeneous genotype, ranging from 99.3% to 100%, within the genospecies of B. burgdorferi sensu stricto and was distinguished from other genospecies of Borrelia isolates. The sequence similarity analysis also revealed the high sequence variability of the ospC gene among Borrelia strains that belong to the same genospecies but were isolated from different biological and geographical sources. Thus, these results provide the first investigation on the genetic identity of the ospC gene of these Taiwan isolates and show that these Taiwan isolates were closely related genetically to the genospecies of B. burgdorferi sensu stricto.