Tags

Type your tag names separated by a space and hit enter

Development of NC1 and NC2 domains of type VII collagen ELISA for the diagnosis and analysis of the time course of epidermolysis bullosa acquisita patients.
J Dermatol Sci. 2011 Jun; 62(3):169-75.JD

Abstract

BACKGROUND

Epidermolysis bullosa acquisita (EBA) is an acquired autoimmune mechanobullous disease. EBA patients possess autoantibodies against type VII collagen which is composed of a collagenous domain flanked by non-collagenous NC1 and NC2 domains. It was reported that major epitopes reside within the NC1 domain and minor epitopes reside within NC2 domain.

OBJECTIVE

The aim of this study is to develop a sensitive and specific ELISA to facilitate the diagnosis of EBA.

METHODS

We developed ELISAs using recombinant NC1 domain produced by mammalian expression system and recombinant NC2 domain produced by mammalian or bacterial expression system to characterize autoantibodies in EBA. Next, we developed an ELISA using a combination of the NC1 (mammalian expression) and NC2 (bacterial expression). We tested the ELISAs with 49 EBA sera, 55 normal control sera, 20 pemphigus vulgaris and 20 bullous pemphigoid sera.

RESULTS

When we evaluated the 49 EBA sera using the NC1 and NC2 ELISAs, 38 (77.5%) reacted with NC1 domain only, 7 sera (14.2%) reacted with both NC1 and NC2 domains, and one serum (2%) reacted with NC2 domain only. Therefore, to increase the sensitivity of the assay, we developed an ELISA coated with a mixture of recombinant NC1 and NC2 domains, resulting in 93.8% sensitivity and 98.1% specificity. By analyzing the time course of two EBA patients, ELISA scores fluctuated in parallel with their disease activity.

CONCLUSION

We conclude that the NC1+NC2 ELISA can be a practical assay for the diagnosis and follow up of the antibody titers of EBA patients.

Authors+Show Affiliations

Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21482078

Citation

Saleh, Marwah Adly, et al. "Development of NC1 and NC2 Domains of Type VII Collagen ELISA for the Diagnosis and Analysis of the Time Course of Epidermolysis Bullosa Acquisita Patients." Journal of Dermatological Science, vol. 62, no. 3, 2011, pp. 169-75.
Saleh MA, Ishii K, Kim YJ, et al. Development of NC1 and NC2 domains of type VII collagen ELISA for the diagnosis and analysis of the time course of epidermolysis bullosa acquisita patients. J Dermatol Sci. 2011;62(3):169-75.
Saleh, M. A., Ishii, K., Kim, Y. J., Murakami, A., Ishii, N., Hashimoto, T., Schmidt, E., Zillikens, D., Shirakata, Y., Hashimoto, K., Kitajima, Y., & Amagai, M. (2011). Development of NC1 and NC2 domains of type VII collagen ELISA for the diagnosis and analysis of the time course of epidermolysis bullosa acquisita patients. Journal of Dermatological Science, 62(3), 169-75. https://doi.org/10.1016/j.jdermsci.2011.03.003
Saleh MA, et al. Development of NC1 and NC2 Domains of Type VII Collagen ELISA for the Diagnosis and Analysis of the Time Course of Epidermolysis Bullosa Acquisita Patients. J Dermatol Sci. 2011;62(3):169-75. PubMed PMID: 21482078.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of NC1 and NC2 domains of type VII collagen ELISA for the diagnosis and analysis of the time course of epidermolysis bullosa acquisita patients. AU - Saleh,Marwah Adly, AU - Ishii,Ken, AU - Kim,Yool-Ja, AU - Murakami,Akihiro, AU - Ishii,Norito, AU - Hashimoto,Takashi, AU - Schmidt,Enno, AU - Zillikens,Detlef, AU - Shirakata,Yuji, AU - Hashimoto,Koji, AU - Kitajima,Yasuo, AU - Amagai,Masayuki, Y1 - 2011/03/16/ PY - 2010/12/21/received PY - 2011/02/28/revised PY - 2011/03/04/accepted PY - 2011/4/13/entrez PY - 2011/4/13/pubmed PY - 2011/9/15/medline SP - 169 EP - 75 JF - Journal of dermatological science JO - J Dermatol Sci VL - 62 IS - 3 N2 - BACKGROUND: Epidermolysis bullosa acquisita (EBA) is an acquired autoimmune mechanobullous disease. EBA patients possess autoantibodies against type VII collagen which is composed of a collagenous domain flanked by non-collagenous NC1 and NC2 domains. It was reported that major epitopes reside within the NC1 domain and minor epitopes reside within NC2 domain. OBJECTIVE: The aim of this study is to develop a sensitive and specific ELISA to facilitate the diagnosis of EBA. METHODS: We developed ELISAs using recombinant NC1 domain produced by mammalian expression system and recombinant NC2 domain produced by mammalian or bacterial expression system to characterize autoantibodies in EBA. Next, we developed an ELISA using a combination of the NC1 (mammalian expression) and NC2 (bacterial expression). We tested the ELISAs with 49 EBA sera, 55 normal control sera, 20 pemphigus vulgaris and 20 bullous pemphigoid sera. RESULTS: When we evaluated the 49 EBA sera using the NC1 and NC2 ELISAs, 38 (77.5%) reacted with NC1 domain only, 7 sera (14.2%) reacted with both NC1 and NC2 domains, and one serum (2%) reacted with NC2 domain only. Therefore, to increase the sensitivity of the assay, we developed an ELISA coated with a mixture of recombinant NC1 and NC2 domains, resulting in 93.8% sensitivity and 98.1% specificity. By analyzing the time course of two EBA patients, ELISA scores fluctuated in parallel with their disease activity. CONCLUSION: We conclude that the NC1+NC2 ELISA can be a practical assay for the diagnosis and follow up of the antibody titers of EBA patients. SN - 1873-569X UR - https://www.unboundmedicine.com/medline/citation/21482078/Development_of_NC1_and_NC2_domains_of_type_VII_collagen_ELISA_for_the_diagnosis_and_analysis_of_the_time_course_of_epidermolysis_bullosa_acquisita_patients_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0923-1811(11)00085-5 DB - PRIME DP - Unbound Medicine ER -