IMMUNOGLOBULIN-M CAPTURE ASSAY FOR SEROLOGIC CONFIRMATION OF EARLY LYME-DISEASE - ANALYSIS OF IMMUNE-COMPLEXES WITH BIOTINYLATED BORRELIA-BURGDORFERI SONICATE ENHANCED WITH FLAGELLIN PEPTIDE EPITOPE
M. Brunner et al., IMMUNOGLOBULIN-M CAPTURE ASSAY FOR SEROLOGIC CONFIRMATION OF EARLY LYME-DISEASE - ANALYSIS OF IMMUNE-COMPLEXES WITH BIOTINYLATED BORRELIA-BURGDORFERI SONICATE ENHANCED WITH FLAGELLIN PEPTIDE EPITOPE, Journal of clinical microbiology, 36(4), 1998, pp. 1074-1080
We previously reported on the efficacy of the enzyme-linked immunoglob
ulin M capture immune complex (IC) biotinylated antigen assay (EMIBA)
for the seroconfirmation of early Lyme disease and active infection wi
th Borrelia burgdorferi. In earlier work we identified non-cross-react
ing epitopes of a number of B. burgdorferi proteins, including flagell
in, We now report on an improvement in the performance of EMIBA with t
he addition of a biotinylated form of a synthetic non-cross-reacting i
mmunodominant flagellin peptide to the biotinylated B. burgdorferi B31
sonicate antigen source with the avidin-biotinylated peroxidase compl
ex: detection system used in our recently developed indirect IgM-captu
re immune complex-based assay (EMIBA). As in our previous studies, the
enzyme-linked immunosorbent assay (ELISA) reactivities of antibodies
liberated from circulating ICs (by EMIBA) were compared with those of
antibodies in unprocessed serum (antibodies found free in the serum, t
hus as an IgM-capture ELISA, but not EMIBA, because the antibodies wer
e not liberated from ICs), the sample usually used in standard ELISAs
and Western blot assays, The addition of the flagellin epitope enhance
d the ELISA signal obtained with untreated sera from many Lyme disease
patients but not from healthy controls, In tests with both free antib
odies and ICs, with or without the addition of the flagellin epitope t
o the sonicate, we found the most advantageous combination was IC as t
he source of antibodies and sonicate plus the flagellin epitope as the
antigen, In a blinded study of sera obtained from patients with early
and later-phase Lyme disease, EMIBA with the enhanced antigenic prepa
ration compared favorably with other serologic assays, especially for
the confirmation of early disease.