Paper-based ELISA for the detection of autoimmune antibodies in body fluid-the case of bullous pemphigoid.
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.
Department of Dermatology, National Cheng Kung University College of Medicine and Hospital , 138 Sheng-Li Road, Tainan 704, Taiwan.
Chao-Kai
CK
Hsu
Hsin-Yu
HY
Huang
Wan-Rung
WR
Chen
Wataru
W
Nishie
Hideyuki
H
Ujiie
Ken
K
Natsuga
Shu-Ting
ST
Fan
Hsi-Kai
HK
Wang
Julia Yu-Yun
JY
Lee
Wei-Lun
WL
Tsai
Hiroshi
H
Shimizu
Chao-Min
CM
Cheng
D001323:Autoantibodies / Q000032:analysis*; D001826:Body Fluids / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D010209:Paper; D010391:Pemphigoid, Bullous / Q000276:immunology*
D001323:Autoantibodies