Post by LymeEnigma on Mar 1, 2008 9:26:00 GMT -8
Serodiagnosis of Lyme Borreliosis by Western Immunoblot:
Reactivity of Various Significant Antibodies against Borrelia burgdorferi
BINGNAN MA, BEYAT CHRISTEN, DANTON LEUNG, AND CARMEN VIGO-PELFREY
Abstract:
The significance of various antibodies against Borrelia burgdorferi was studied by Western blot (immunoblot) by using 578 human serum samples. The proteins regularly detected by using samples from patients with Lyme borreliosis were those with bands with molecular masses of 94, 83, 75, 66, 60, 55, 46, 41, 39, 34, 31, 29, 22, and 17 kDa. The detectable frequencies of most of these proteins appeared to be significantly different between the sera from patients with Lyme borreliosis and those from normal control individuals as well as from the group with syphilis. The 39-kDa protein band recognized by polyvalent antibody was found to be the most significant marker for Lyme borreliosis. Furthermore, an anti-39-kDa immunoglobulin M response was detected in the samples from patients with early-stage Lyme borreliosis. Results from the use of monoclonal antibodies and patient sera revealed that the 39- and 41-kDa proteins may be structurally related but are immunologically distinct antigens. The significance of antibody reactivities to the 41-, 94-, 22-, 31-, and 34-kDa protein bands is also discussed.
Full article: www.pubmedcentral.nih.gov/picrender.fcgi?artid=265062&blobtype=pdf
*special note: Nick Harris, then of Whittaker Bioproducts, was an adviser in the accruing of information used in this paper.
Comparative Evaluation of Three Products for the Detection of
Borrelia burgdorferi Antibody in Human Serum
R. D. FISTER, L. A. WEYMOUTH,2 J. C. McLAUGHLIN, R. W. RYAN, AND R. C. TILTON
Abstract:
Eighty human serum specimens tested concomitantly by immunoblot and an enzyme-linked immunosorbent assay developed jointly at the University of Connecticut School of Medicine and the Connecticut Agricultural Experiment Station were used to evaluate three commercially available diagnostic products for Lyme borreliosis. The sources of the kits were Hillcrest Biologicals, Cypress, Calif.; Whittaker Bioproducts, Walkersville, Md.; and Cambridge Bioscience, Worcester, Mass. When compared with Western blot analysis, the sensitivities and specificities, respectively, for the diagnostic assays were as follows: Hillcrest Biologicals, 93 and 75%; Whittaker Bioproducts, 73 and 100%; Cambridge Bioscience, 89 and 100%; and University of Connecticut School of Medicine, 96 and 92%.
Full article: www.pubmedcentral.nih.gov/picrender.fcgi?artid=267136&blobtype=pdf
Western blot analysis of sera from Lyme borreliosis patients according to the genomic species of the Borrelia strains used as antigens
Abstract:
Sera of 52 Lyme borreliosis patients classified according to their clinical features were analysed by Western blot using as antigens Borrelia strains belonging to three recently described genomic species. The antibody response was demonstrated to be homologous within each genospecies. Serum reactivity was studied for each of the type strains Borrelia burgdorferi sensu stricto (strain B31T), Borrelia garinii (strain 20047T) and group VS461. Seven of 15 sera (46.6 %) of patients with menin-goradiculitis showed preferential reactivity with Borrelia garinii (strain 20047T), all of 8 sera (100 %) of patients with acrodermatitis chronica atrophicans showed preferential reactivity with group VS461 (strain VS461) and 8 of 16 sera (50 %) of patients with arthritis showed preferential reactivity with Borrelia burgdorferi sensu stricto (strain B31T). The presence of a strong response to OspA and OspB proteins of Borrelia burgdorferi sensu stricto was found only in this last group of patients. These results suggest that there are clinical implications of the recently described modifications in the taxonomy of Borrelia burgdorferi. www.springerlink.com/content/v2176801411q4512/
Partial breakdown of bands
(from Laboratory aids for the diagnosis of Borrelia burgdorferi infection, by Richard C. Tilton, Ph.D.):
1. 83 Kda. LeFebvre et al. (26) suggested that this chromosomally expressed protein is genus-species specific. Dorward (27) confirmed
the species specificity of this antigen.
2. 39 Kda. A major protein determinant in B. burgdorferi (21).
3. 34 Kda protein. Outer surface protein B.
4. 31 Kda protein. Outer surface protein A.
5. 25/22 Kda protein. Outer surface protein C (28).
6. 12 Kda protein. Genus-species specific but function unknown (29).
7. 41 Kda. This protein is characteristic of the flagellum of B. burgdorferi, but also of other treponemes such as oral treponemes that are involved in periodontal disease, other borrelias, and occasionally Leptospira. IgM antibody to 41 Kda is usually the first antibody to appear after infection with B. burgdorferi.
It is our experience and that of others that patients with latestage or recurrent Lyme disease may develop a variety of antibodies to other proteins (9, 20, 37, 38, 45, 50, 55, 57, 60, 66, 75, 100 Kda) but similar patterns may occasionally be seen in control subjects.
The significance of these proteins is unknown. The interpretation of immunoblots for Lyme disease should be based on both the number and type of bands present. We do not feel that interpretation based solely on the number of bands without regard to the nature of the proteins is justified. For example, more emphasis is placed on antibodies to proteins such as 22, 25 Kda, OspA (31 Kda), OspB (34 Kda), 39 Kda, and 83 Kda than the cross-reactive proteins such as 41 Kda or those with undefined significance. While cross-reactive, antibody to the 41 Kda protein is still important for the detection of early Lyme disease.
There are three categories of antibodies observed in Western blots:
Category 1- known cross-reactive and other undefined bands (9, 20, 37, 38, 45, 50, 55, 57, 60, 66 Kda);
Category 2- genus-family specific (41 Kda);
Category 3- genus-species specific (12, 22, 25, 3 1, 34, 39, 83 Kda).
www.jstd.org/journal/vol1no1/v1n1_lab.pdf
Recommendation To Include OspA and OspB in the New Immunoblotting Criteria for Serodiagnosis of Lyme Disease: jcm.asm.org/cgi/reprint/34/6/1353.pdf
Reactivity of Various Significant Antibodies against Borrelia burgdorferi
BINGNAN MA, BEYAT CHRISTEN, DANTON LEUNG, AND CARMEN VIGO-PELFREY
Abstract:
The significance of various antibodies against Borrelia burgdorferi was studied by Western blot (immunoblot) by using 578 human serum samples. The proteins regularly detected by using samples from patients with Lyme borreliosis were those with bands with molecular masses of 94, 83, 75, 66, 60, 55, 46, 41, 39, 34, 31, 29, 22, and 17 kDa. The detectable frequencies of most of these proteins appeared to be significantly different between the sera from patients with Lyme borreliosis and those from normal control individuals as well as from the group with syphilis. The 39-kDa protein band recognized by polyvalent antibody was found to be the most significant marker for Lyme borreliosis. Furthermore, an anti-39-kDa immunoglobulin M response was detected in the samples from patients with early-stage Lyme borreliosis. Results from the use of monoclonal antibodies and patient sera revealed that the 39- and 41-kDa proteins may be structurally related but are immunologically distinct antigens. The significance of antibody reactivities to the 41-, 94-, 22-, 31-, and 34-kDa protein bands is also discussed.
Full article: www.pubmedcentral.nih.gov/picrender.fcgi?artid=265062&blobtype=pdf
*special note: Nick Harris, then of Whittaker Bioproducts, was an adviser in the accruing of information used in this paper.
Comparative Evaluation of Three Products for the Detection of
Borrelia burgdorferi Antibody in Human Serum
R. D. FISTER, L. A. WEYMOUTH,2 J. C. McLAUGHLIN, R. W. RYAN, AND R. C. TILTON
Abstract:
Eighty human serum specimens tested concomitantly by immunoblot and an enzyme-linked immunosorbent assay developed jointly at the University of Connecticut School of Medicine and the Connecticut Agricultural Experiment Station were used to evaluate three commercially available diagnostic products for Lyme borreliosis. The sources of the kits were Hillcrest Biologicals, Cypress, Calif.; Whittaker Bioproducts, Walkersville, Md.; and Cambridge Bioscience, Worcester, Mass. When compared with Western blot analysis, the sensitivities and specificities, respectively, for the diagnostic assays were as follows: Hillcrest Biologicals, 93 and 75%; Whittaker Bioproducts, 73 and 100%; Cambridge Bioscience, 89 and 100%; and University of Connecticut School of Medicine, 96 and 92%.
Full article: www.pubmedcentral.nih.gov/picrender.fcgi?artid=267136&blobtype=pdf
Western blot analysis of sera from Lyme borreliosis patients according to the genomic species of the Borrelia strains used as antigens
Abstract:
Sera of 52 Lyme borreliosis patients classified according to their clinical features were analysed by Western blot using as antigens Borrelia strains belonging to three recently described genomic species. The antibody response was demonstrated to be homologous within each genospecies. Serum reactivity was studied for each of the type strains Borrelia burgdorferi sensu stricto (strain B31T), Borrelia garinii (strain 20047T) and group VS461. Seven of 15 sera (46.6 %) of patients with menin-goradiculitis showed preferential reactivity with Borrelia garinii (strain 20047T), all of 8 sera (100 %) of patients with acrodermatitis chronica atrophicans showed preferential reactivity with group VS461 (strain VS461) and 8 of 16 sera (50 %) of patients with arthritis showed preferential reactivity with Borrelia burgdorferi sensu stricto (strain B31T). The presence of a strong response to OspA and OspB proteins of Borrelia burgdorferi sensu stricto was found only in this last group of patients. These results suggest that there are clinical implications of the recently described modifications in the taxonomy of Borrelia burgdorferi. www.springerlink.com/content/v2176801411q4512/
Partial breakdown of bands
(from Laboratory aids for the diagnosis of Borrelia burgdorferi infection, by Richard C. Tilton, Ph.D.):
1. 83 Kda. LeFebvre et al. (26) suggested that this chromosomally expressed protein is genus-species specific. Dorward (27) confirmed
the species specificity of this antigen.
2. 39 Kda. A major protein determinant in B. burgdorferi (21).
3. 34 Kda protein. Outer surface protein B.
4. 31 Kda protein. Outer surface protein A.
5. 25/22 Kda protein. Outer surface protein C (28).
6. 12 Kda protein. Genus-species specific but function unknown (29).
7. 41 Kda. This protein is characteristic of the flagellum of B. burgdorferi, but also of other treponemes such as oral treponemes that are involved in periodontal disease, other borrelias, and occasionally Leptospira. IgM antibody to 41 Kda is usually the first antibody to appear after infection with B. burgdorferi.
It is our experience and that of others that patients with latestage or recurrent Lyme disease may develop a variety of antibodies to other proteins (9, 20, 37, 38, 45, 50, 55, 57, 60, 66, 75, 100 Kda) but similar patterns may occasionally be seen in control subjects.
The significance of these proteins is unknown. The interpretation of immunoblots for Lyme disease should be based on both the number and type of bands present. We do not feel that interpretation based solely on the number of bands without regard to the nature of the proteins is justified. For example, more emphasis is placed on antibodies to proteins such as 22, 25 Kda, OspA (31 Kda), OspB (34 Kda), 39 Kda, and 83 Kda than the cross-reactive proteins such as 41 Kda or those with undefined significance. While cross-reactive, antibody to the 41 Kda protein is still important for the detection of early Lyme disease.
There are three categories of antibodies observed in Western blots:
Category 1- known cross-reactive and other undefined bands (9, 20, 37, 38, 45, 50, 55, 57, 60, 66 Kda);
Category 2- genus-family specific (41 Kda);
Category 3- genus-species specific (12, 22, 25, 3 1, 34, 39, 83 Kda).
www.jstd.org/journal/vol1no1/v1n1_lab.pdf
Recommendation To Include OspA and OspB in the New Immunoblotting Criteria for Serodiagnosis of Lyme Disease: jcm.asm.org/cgi/reprint/34/6/1353.pdf