Evaluation of the value of Western blot analysis based on human epidermal extracts prepared by thermal separation in the diagnosis of bullous pemphigoid
10.3760/cma.j.issn.0412-4030.2019.01.002
- VernacularTitle:基于热分离法制备的人表皮侧抗原的Western印迹实验对大疱性类天疱疮的诊断价值评估
- Author:
Suo LI
1
;
Xianfeng CHENG
;
Xinyu LI
;
Zhiliang LI
;
Ke JING
;
Ruiyu XIANG
;
Hanmei ZHANG
;
Suying FENG
Author Information
1. 中国医学科学院北京协和医学院皮肤病医院皮肤科江苏省皮肤病与性病学重点实验室
- Keywords:
Pemphigoid,bullous;
Blotting,Western;
Epidermis;
Autoantigens;
Enzyme-linked immunosorbent assay;
Thermal separation;
Human epidermal extract
- From:Chinese Journal of Dermatology
2019;52(1):6-10
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare human epidermal extracts by thermal separation,and to evaluate the value of epidermal extract-based Western blot analysis in the diagnosis of bullous pemphigoid (BP).Methods Human epidermal extracts were prepared by thermal separation from circumcised foreskins of healthy males.Serum samples were obtained from 22 inpatients with BP and 25 inpatients without BP in Hospital for Skin Diseases,Chinese Academy of Medical Sciences and Peking Union Medical College between January 2015 and August 2017.These serum samples were subjected to Western blot analysis with epidermal extracts as substrates,as well as to BP180-NC16A enzyme-linked immunosorbent assay (ELISA).Statistical analysis was carried out using chi-square test and Fisher's exact test with the SPSS22.0 software.Results The sensitivities of epidermal extract-based Western blot analysis and BP 180-NC16A ELISA in the diagnosis of BP were 86.36％ (95 ％ CI:64.03％-96.41％) and 95.45％ (95％ CI:75.11％-99.76％) respectively (~ =1.10,P =0.294),and the specificities were 100％ (95％ CI:83.42％-100％) and 92％ (95％ CI:75.11％-99.76％) respectively (x2 =20.8,P =0.149).Epidermal extract-based Western blot analysis in the 22 patients with BP showed a protein band with relative molecular mass (RMM) of 230 000 in 4 patients,a protein band with RMM of 180 000 in 18,a protein band with RMM of 120 000 in 1,and a protein band with RMM of 97 000 in 1.The BP180-NC16A ELISA showed that the antibody titers were more than 50 U/ml in the BP patients with protein bands of RMM of 180 000.Conclusions The epidermal extract-based Western blot analysis mainly showed the protein band with RMM of 180 000 in the patients with BP.The sensitivity of the epidermal extract-based Western blot analysis was lower than that of the BP180-NC16A ELISA,and the epidermal extract-based Western blot analysis tends to be negative when the titer of the autoantibody is low.
