Paper-based ELISA for the detection of autoimmune antibodies in body fluid-the case of bullous pemphigoid

Chao Kai Hsu, Hsin Yu Huang, Wan Rung Chen, Wataru Nishie, Hideyuki Ujiie, Ken Natsuga, Shu Ting Fan, Hsi Kai Wang, Julia Yu Yun Lee, Wei Lun Tsai, Hiroshi Shimizu, Chao Min Cheng

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66 Citations (Scopus)


Bullous pemphigoid (BP), a common autoimmune blistering disease, is increasing in incidence and conveys a high mortality. Detection of autoantibodies targeting the noncollagenous 16A (NC16A) domain of type XVII collagen using enzyme-linked immunosorbent assay (ELISA) has demonstrated high sensitivity and specificity for diagnosing BP. We have developed a rapid, low-cost, and widely applicable ELISA-based system to detect the NC16A autoimmune antibody and then diagnose and monitor BP disease activity using a piece of filter paper, a wax-printer, and NC16A antigens. Both sera and/or blister fluids from 14 untreated BP patients were analyzed. The control group included healthy volunteers and patients with other blistering disorders such as pemphigus vulgaris. In our established paper-based ELISA (P-ELISA) system, only 2 μL of serum or blister fluid and 70 min were required to detect anti-NC16A autoimmune antibodies. The relative color intensity was significantly higher in the BP group than in the control groups when using either serum (P < 0.05) or blister fluid (P < 0.001) specimens from BP patients. The results of P- ELISA were moderately correlated with the titer of the commercial ELISA kit (MBL, Japan) (rho = 0.5680, P = 0.0011). This newly developed system allows for rapid and convenient diagnosis and/or monitoring of BP disease activity.

Original languageEnglish
Pages (from-to)4605-4610
Number of pages6
JournalAnalytical chemistry
Issue number9
Publication statusPublished - 2014 May 6

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

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