Abstract | BACKGROUND: OBJECTIVES: To develop a high-performance immunoblotting method using human amniotic membrane as the antigen source, and to compare it with current laboratory methods. METHODS: RESULTS: In BP, the three methods had similar sensitivity (84-89%) for both anti-BP180-NC16a and anti-BP230 antibody detection. In MMP, autoantibodies (mainly directed against BP180 or laminin 332 subunits) were detected in 77% of patients by IB, compared with only 9% by IIF on rat and monkey oesophagus and 36% on salt-split skin, and 14% by anti-BP180-NC16a and anti-BP230 ELISA. In patients with pemphigus, ELISA had 92% sensitivity for anti-Dsg1 and 3, but IB and rat bladder IIF were necessary to confirm PNP by revealing specific and rare patterns (antidesmoplakin I/II, antienvoplakin and antiperiplakin antibodies). IB also revealed anticollagen VII antibodies in 60% of patients with EBA and BSLE, and antibodies to BP180, BP230 and Dsg3 in a few patients who were negative using the other two techniques. CONCLUSION: Amniotic membrane immunoblotting is an interesting diagnostic tool for bullous diseases, as the entire panel of autoantibodies can be detected with a single extract. This method improves the identification of complex and heterogeneous autoimmune processes in conjunction with IIF and ELISA, and is particularly useful for MMP characterization.
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Authors | S Grootenboer-Mignot, V Descamps, C Picard-Dahan, P Nicaise-Roland, C Prost-Squarcioni, C Leroux-Villet, C Champagnat, A Delaval, F Aucouturier, B Crickx, S Chollet-Martin |
Journal | The British journal of dermatology
(Br J Dermatol)
Vol. 162
Issue 4
Pg. 743-50
(Apr 2010)
ISSN: 1365-2133 [Electronic] England |
PMID | 19886889
(Publication Type: Journal Article)
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Chemical References |
- Autoantibodies
- Biomarkers
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Topics |
- Amnion
(immunology)
- Animals
- Autoantibodies
(immunology)
- Biomarkers
- Case-Control Studies
- Enzyme-Linked Immunosorbent Assay
(methods)
- Fluorescent Antibody Technique
(methods)
- Haplorhini
- Humans
- Immunoblotting
(methods)
- Rats
- Skin Diseases, Vesiculobullous
(diagnosis, immunology)
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