Western blotting in the serodiagnosis of Lyme disease.
Dressler F, Whalen JA, Reinhardt BN, Steere AC. Division of Rheumatology/Immunology, Tufts University School of Medicine, New England Medical Center, Boston, Massachusetts 02111.
There are currently no accepted criteria for positive Western blots in Lyme disease.
In a retrospective analysis of 225 case and control subjects, the best discriminatory ability of test criteria was obtained by requiring at least 2 of
the 8 most common IgM bands in early disease
(18, 21, 28, 37, 41, 45, 58, and 93 kDa)
and by requiring at least 5 of
the 10 most frequent IgG bands after the first weeks of infection
(18, 21, 28, 30, 39, 41, 45, 58, 66, and 93 kDa)
When these definitions were tested in a prospective study of all 237 patients seen in a diagnostic Lyme disease clinic during a 1-year period and in 74 patients with erythema migrans or summer flu-like illnesses, the IgM blot in early disease had a sensitivity of 32% and a specificity of 100%; the IgG blot after the first weeks of infection had a sensitivity of 83% and a specificity of 95%.
Among patients with indeterminate IgG responses by ELISA, 6 of 9 patients with active Lyme disease had positive blots compared with 2 of 34 patients with other illnesses (P < .001). Thus, Western blotting can be used to increase the specificity of serologic testing in Lyme disease.
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Electrophoresis. 1993 Sep;14(9):937-44.Links Western blot as a tool in the diagnosis of Lyme borreliosis.Zöller L, Cremer J, Faulde M. Department of Medical Microbiology, Ernst-Rodenwaldt-Institute, Koblenz, Germany.
Borrelia burgdorferi is the causative agent of Lyme borreliosis, a multisystem disorder, which can mimic numerous immune disorders and inflammatory diseases. Laboratory diagnosis of Borrelia infection relies on immunodiagnostic assays, which, however, are hampered by unsatisfactory specificity. The Western blot technique has been employed to analyze the humoral immune response in Lyme borreliosis and is used as a serodiagnostic confirmation test.
The most important immunodominant proteins of Borrelia burgdorferi are the 94 kDa, 60 kDa, 41 kDa (flagellin), 34 kDa (Osp B), 31 kDa (Osp A), 30 kDa, 21 kDa (Osp C), and 17/18 kDa proteins.
Whereas the 60 kDa, 41 kDa, and 34 kDa constituents reveal a marked cross-antigenicity with other spirochetes and even more distantly related bacteria, antibodies against the 94 kDa, 31 kDa and 21 kDa proteins are largely species-specific.
The early immune response in Lyme borreliosis is triggered mainly by the flagellin. In the later stage a wide range of immunogenic proteins is involved, with the 94 kDa antigen being the best marker for late immune response. If the Western blot is used for diagnostic purposes the differences between early and late-stage immunogenicity of Borrelia proteins must be taken into account.
Interpretation criteria for blot positivity in early-stage borreliosis are primarily based on the presence of the 21 kDa band and the semiquantitatively recorded intensity of the 41 kDa band. In the diagnosis of late-stage infection, blot positivity relies on the presence of the 94 kDa, 39 kDa, 31 kDa, 30 kDa and 21 kDa bands.
PMID: 8223404 [PubMed - indexed for MEDLINE]
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IgE anti-Borrelia burgdorferi components (p18, p31, p34, p41, p45, p60) and increased blood CD8+CD60+ T cells in children with Lyme disease.
Bluth MH, Robin J, Ruditsky M, Norowitz KB, Chice S, Pytlak E, Nowakowski M, Durkin HG, Smith-Norowitz TA. Department of Surgery, S.U.N.Y. Downstate Medical Center, Brooklyn, NY, USA.
Immunoglobulin (Ig) E may provide immunity against Borrelia burgdorferi infection (Lyme disease) in children which lasts throughout adulthood. We investigated the presence and persistence of IgE anti-B. burgdorferi antibodies (Abs) in paediatric patients infected with Lyme disease over time. Serum immunoglobulin levels, presence of IgG and IgE anti-B. burgdorferi components, and distributions of blood T, B and natural killer lymphocyte subsets were studied in B. burgdorferi-infected and -uninfected children (nephelometry, UniCAP Total IgE Fluoroenzymeimmunoassay, Western blot, flow cytometry). Total serum IgM, IgG, IgE and IgA levels, and distributions of blood lymphocytes (CD4(+), CD8(+), CD19(+)) of both groups, excluding CD8(+)CD60(+) T cells, were within normal ranges.
However, infected, but not uninfected children made IgG anti-B. burgdorferi proteins p18, p31, p34, p41, p45, but not IgG anti-p60, and IgE anti-B. burgdorferi proteins p31, p34, p41, p45, p60, but not IgE anti-p18.
These proteins were also detected in an infected child 1 year post-infection. Interestingly, CD8(+)CD60(+) T-cell numbers were significantly increased (fourfold) in infected, compared with uninfected, patients (P=0.001).
These results demonstrate that specific IgE anti-B. burgdorferi Abs are generated and persist in children with Lyme disease and that CD8(+)CD60(+) T cells may play an important role in these responses.
PMID: 17386029 [PubMed - indexed for MEDLINE]
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Complement component (3d/Epstein Barr virus) receptor 2, also known as CR2 and CD21, is a protein involved in the complement system. It binds to C3d. B cells have CR2 receptors on their surfaces, allowing the complement system to play a role in B-cell activation and maturation 
Complement component receptor-2 (CR2) is the membrane protein on B lymphocytes to which the Epstein-Barr virus (EBV) binds during infection of these cells. See also MIM 120620. Yefenof et al. (1976) found complete overlapping of EBV receptors and C3 (MIM 120700) receptors on human B lymphocytes.[supplied by OMIM]
They think he has Epstein-Barr virus But that cross reacts with Lyme.
18 kDa p18: the specificity of this protein has been recently established; excellent marker for late infections.
It is therefore often useful to include the p18 band in the interpretation, knowing that it will be found in only 25-30% of EM but is virtually never present on blots from healthy patients. Here Band 18
Results. Lyme arthritis patients had more bands and higher-intensity bands than did non-Lyme patients. The presence of at least 2 bands of moderate to high intensity (>40 optical units) or at least 5 bands of lower intensity (>20 optical units) was over 90% sensitive and 100% specific for the diagnosis of Lyme arthritis. A 60-kd band was present in all Lyme arthritis patients. The presence of an 83-, 39-, 21-, or 18-kd band was highly specific for Lyme arthritis.
The presence of an 83-, 39-, 21-, or 18-kd band was highly specific for Lyme arthritis. CONCLUSION. Band intensity analysis increases the objectivity and accuracy of Western blot interpretation for the diagnosis of Lyme arthritis. here pubmedband 18
The overall reactivity of the three Borrelia strains in IgG immunoblots indicated that ten protein bands were significant, with a different prevalence of some of them in the two groups of patient sera: bands at 60-58, 30-33, 36-37 and 28-27 kDa were markers for neuroborreliosis sera; proteins at 100-83, 72-70 and 18-17 kDa behaved like markers for Lyme arthritis.
Conclusions: The CDC guidelines result in a gross underreporting of true Lyme cases, as well as the misdiagnosis of many Lyme sufferers. More reliable testing is needed, and the CDC needs to lower the number of Western Blot bands required, also adding the Bb-specific bands of 31 kDa, 34 kDa, and 18kDa. here canlyme 18kda
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