[Isolation of the recombinant proteins OspC and the fragment FlaA (F-FLAA) from the western-Siberian Borrelia garinii NT29 isolates and the study of their immunochemical properties].
The structural proteins OspC and FlaA of the Lyme disease (LD) agent are known to be the basic antigens, which induce the humoral immune response at the initial stage of the disease. The goal of this work was to obtain the recombinant OspC and a fragment of the FlaA protein (f-FlaA) from the Western Siberian Borrelia garinii NT29 isolates and to assess the possibility of their use for the LD diagnosis. Encoding regions of the OspC and f-FlaB genes were amplified using PCR inserted in the pREB expressive vectors and cloned in the E. coli str. BL21 and C-600, respectively. The recombinant OspC and f-FlaA proteins were purified using affinity chromatography on Ni-NTA-sepharose 6A, and their ability to bind serum antibody of patients with Lyme disease was tested using western-blot and ELISA methods. The results of the analyses suggest that these proteins can be considered as promising components for elaboration of diagnostic tests for LD. The prototype of the ELISA diagnostic test was designed on the basis of the OspC and f-FlaA recombinant antigens. This test provides satisfactory parameters of diagnostic specificity (70.0%) and sensitivity (85.0%).
Karavaev
V S
VS
Ivanov
I D
ID
Riabchenko
A V
AV
Beklemishev
A B
AB
D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D005408:Flagellin / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D008193:Lyme Disease / Q000175:diagnosis* / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012800:Siberia
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C079616:OspC protein; D011994:Recombinant Proteins; D005408:Flagellin; C067138:flaA protein, bacteria