Rapid immunochromatographic test for detection of anti-factor XIII A subunit antibodies can diagnose 90 % of cases with autoimmune haemorrhaphilia XIII/13.
Autoimmune haemorrhaphilia XIII/13 (AH13) is an acquired life-threatening bleeding disorder due to anti-factor XIII (FXIII) autoantibodies (auto-Abs). AH13 patients may die of haemorrhage without correct diagnosis and proper treatment because of lack of awareness and the absence of rapid easy-to-use tests specific for this disease. Currently, the definitive diagnosis is established by cumbersome and time-consuming laboratory tests such as dot-blot assays and enzyme-linked immunosorbent assays (ELISA), and therefore these tests are generally not carried out. To save AH13 patients' lives, there is an urgent necessity for developing a rapid test for FXIII auto-Abs. We first generated and characterised mouse monoclonal antibodies (mAb) against human FXIII A subunit (FXIII-A), and then developed a rapid immunochromatographic test (ICT) for detection of anti-FXIII-A auto-Abs using one mAb with a dissociation constant of 9.3 × 10⁻¹¹ M. The auto-Ab-FXIII-A complex was captured by the mAb on a nitrocellulose membrane and visualised by Au-conjugated anti-human IgG Ab. Mixing with healthy control plasma improved the detection of auto-Abs in patients having extremely low levels of FXIII-A. The specificity and sensitivity of the ICT were 87 % and 94 %, respectively. We also detected auto-Abs against activated FXIII (FXIIIa) in three patients by pre-converting FXIII to FXIIIa by thrombin treatment. ICT values were significantly inversely correlated with FXIII activity levels, indicating an association between the quantity of anti-FXIII autoantibodies and AH13. This reliable rapid ICT assay can be applied to a point-of-care test to detect anti-FXIII-A auto-Abs, and will contribute to early diagnosis and treatment of AH13.
Tsukaka
T
Osaki
Daisuke
D
Sugiyama
Yasuo
Y
Magari
Masayoshi
M
Souri
Akitada Ichinose, MD, PhD, Department of Molecular Patho-Biochemistry and Patho-Biology, Yamagata University School of Medicine, 2-2-2 Iida-Nishi, Yamagata, 990-9585 Japan, Tel.: +81 23 628 5276, Fax: +81 23 628 5280, E-mail: aichinos@med.id.yamagata-u.ac.jp.
Akitada
A
Ichinose
D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology; D000918:Antibody Specificity; D019540:Area Under Curve; D001323:Autoantibodies / Q000097:blood*; D001327:Autoimmune Diseases / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D002138:Calibration; D016022:Case-Control Studies; D002846:Chromatography, Affinity / Q000379:methods* / Q000592:standards; D042241:Early Diagnosis; D018604:Epitope Mapping; D005176:Factor XIII / Q000276:immunology*; D005177:Factor XIII Deficiency / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D000067716:Point-of-Care Testing / Q000592:standards; D011237:Predictive Value of Tests; D012372:ROC Curve; D012015:Reference Standards; D015203:Reproducibility of Results
D058846:Antibodies, Monoclonal, Murine-Derived; D001323:Autoantibodies; C064015:factor XIII subunit A; D005176:Factor XIII
Acquired coagulation disorders; autoantibodies; autoimmune diseases; diagnosis management; factor XIII / transglutaminases