Orientia tsutsugamushi, a causative pathogen of Scrub typhus, is a gram-negative intracellular bacterium. Outer membrane vesicles (OMVs) are produced from the membrane of bacteria and play many roles related to the survival of the pathogen. However, there have been no reports confirming whether O. tsutsugamushi indeed produce OMVs. O. tsutsugamushi boryong was cultured in ECV-304 cells for the purification of OMVs. Western blot analysis and immunoenrichment using anti-O. tsutsugamushi monoclonal antibody and electron microscopy were employed for identification and characterization of OMVs. We confirm the presence of OMVs derived from O. tsutsugamushi, and also found that those OMVs contain a major surface antigen of 56-kDa protein and variant immunogenic antigens.
Antibodies, Monoclonal/*immunology ; Antigens, Bacterial/*immunology ; Antigens, Surface/*immunology ; Cell Line ; Cell Membrane/immunology ; Humans ; Microscopy, Electron ; Orientia tsutsugamushi/*immunology/metabolism ; Scrub Typhus/diagnosis/microbiology ; Secretory Vesicles/*immunology

There are several antigenic variants of Orientia tsutsugamushi. The 56-kDa type-specific antigen (TSA) is responsible for the antigenic variation. Nucleotide sequences of the 56-kDa TSA obtained from 44 eschar samples of Korean scrub typhus patients and from 40 representative strains retrieved from the GenBank database were analyzed phylogenetically. Clinical patient data were assessed based on the genotyping results. Of the 44 nucleotide sequences, 32 (72.7%) clustered with the Boryong genotype, which is the major genotype in Korea. Eleven nucleotide sequences (25%) clustered with the Kawasaki genotype, not identified in Korea until 2010. One nucleotide sequence was consistent with the Karp genotype. The clinical course of the patients infected with each genotype showed no differences. Diagnostic performance of the immunofluorescence assay (IFA) using the 56-kDa TSA from Gilliam, Karp and Boryong as test antigens were not different for the Boryong and Kawasaki genotypes. Although Boryong is still the predominant genotype, the results suggest that Kawasaki genotype is quite prevalent in Chungbuk province of Korea.
Aged ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; DNA, Bacterial/analysis ; Databases, Genetic ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Open Reading Frames ; Orientia tsutsugamushi/*classification/isolation & purification ; Phylogeny ; Republic of Korea ; Scrub Typhus/diagnosis/metabolism/*microbiology ; Sequence Analysis, DNA

There are several antigenic variants of Orientia tsutsugamushi. The 56-kDa type-specific antigen (TSA) is responsible for the antigenic variation. Nucleotide sequences of the 56-kDa TSA obtained from 44 eschar samples of Korean scrub typhus patients and from 40 representative strains retrieved from the GenBank database were analyzed phylogenetically. Clinical patient data were assessed based on the genotyping results. Of the 44 nucleotide sequences, 32 (72.7%) clustered with the Boryong genotype, which is the major genotype in Korea. Eleven nucleotide sequences (25%) clustered with the Kawasaki genotype, not identified in Korea until 2010. One nucleotide sequence was consistent with the Karp genotype. The clinical course of the patients infected with each genotype showed no differences. Diagnostic performance of the immunofluorescence assay (IFA) using the 56-kDa TSA from Gilliam, Karp and Boryong as test antigens were not different for the Boryong and Kawasaki genotypes. Although Boryong is still the predominant genotype, the results suggest that Kawasaki genotype is quite prevalent in Chungbuk province of Korea.
Aged ; Bacterial Proteins/*genetics/metabolism ; Base Sequence ; DNA, Bacterial/analysis ; Databases, Genetic ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Open Reading Frames ; Orientia tsutsugamushi/*classification/isolation & purification ; Phylogeny ; Republic of Korea ; Scrub Typhus/diagnosis/metabolism/*microbiology ; Sequence Analysis, DNA