OBJECTIVETo recombine OspC gene from Borrelia burgdorferi PD91 of China and expressed it in E. coli for early diagnosis of Lyme disease.

METHODSThe OspC gene was amplified from the genome of Borrelia burgdorferi PD91 strain by polymerase chain reaction and recombined with plasmid PET-11D. The recombinant plasmid PET-11D-OspC was identified with PCR, restriction endonuclease analysis and sequencing. The antigenicity was verified with Western Blot.

RESULTSOspC gene was cloned correctly into vector PET-11D. The resultant sequence was definitely different from the published sequence. The recombinant OspC seemed to have had strong antigenicity.

CONCLUSIONThe findings laid basis for the studies on early diagnosis of Lyme disease.

Antigens, Bacterial ; Bacterial Outer Membrane Proteins ; genetics ; immunology ; Blotting, Western ; Borrelia burgdorferi Group ; immunology ; Escherichia coli ; genetics ; Humans ; Lyme Disease ; diagnosis ; Polymerase Chain Reaction ; Recombinant Proteins ; analysis ; immunology

Lyme disease, a multi-systemic infection occurring worldwide, has yet to be reported in Korea, although the spirochete B. burgdorferi, known as the causative organism of the disease, has recently been isolated from the vector tick Ixodes persulcatus in the region. To contribute to revealing whether Lyme disease exists in Korea or not, B. burgdorferi specific antibodies (IgG, IgM, and/or IgA) were measured by three individual enzyme-linked immunosorbent assays (ELISA) utilizing different antigens in 38 patients with juvenile rheumatoid arthritis (JRA) which shares a number of clinical features with Lyme arthritis. The antibody prevalence rates in patients with JRA were various depending on the antigens (21% of IgG and IgM antibodies to purified organisms, 0% for IgG antibody to purified native flagella, and 5% for IgG, IgM, and IgA antibodies to recombinant p39) and were not different compared to 39 controls (21%, 0%, and 0% respectively). The antibody prevalence rates compared in various subgroups of patients with JRA according to types of JRA, length of illness, age, and sex were not different. Comparing the three different antigens, the greatest number of positive responders were yielded by purified organisms followed by p39 and purified flagellin, however the possibility of nonspecificity with purified organisms remained. The data indicate that serologic tests using ELISA fail to illustrate Lyme disease among 38 patients with JRA in Korea.
Adolescent ; Adult ; Antibodies, Bacterial/*blood ; Arthritis, Juvenile Rheumatoid/*microbiology ; Borrelia burgdorferi Group/*immunology ; Child ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male