Rapid quantitative assay of ADAMTS13 activity on an automated coagulation analyzer: clinical applications and comparison with immunoblot method.
Facile assays for ADAMTS13, the metalloprotease that is absent or impaired in thrombotic thrombocytopenic purpura, could allow for timely diagnosis and management of this potentially fatal hematologic disorder; unfortunately, available assays employ methodologies that restrict use to only a few reference laboratories, resulting in reporting delays. Measurement of ADAMTS13 on an automated analyzer could allow widespread accessibility by routine clinical laboratories. We adapted a previously published technique to demonstrate that ADAMTS13 activity could be measured on an Instrumentation Laboratory automated coagulation analyzer after plasma digestion of a commercial source of von Willebrand factor (VWF). Results were obtained using a commercially available immunoturbidimetric assay for residual VWF activity. Samples from 114 patients with suspected thrombotic microangiopathy were analyzed by both standard immunoblot and our new semiautomated method with excellent agreement, particularly at the clinically relevant low levels of ADAMTS13 activity. This new method is semiautomated and offers quantitative results within 2 hr; our modifications allow for widespread applicability on instrumentation already in use by routine clinical laboratories.
Department of Internal Medicine, Wake Forest University Health Sciences, Winston-Salem, North Carolina 27157, USA. mknovich@wfubmc.edu
Mary Ann
MA
Knovich
Heather L
HL
Lawson
Martha H
MH
Burke
Thomas P
TP
McCoy
John
J
Owen
D051722:ADAM Proteins / Q000097:blood*; D000071120:ADAMTS13 Protein; D001331:Automation; D001780:Blood Coagulation Tests / Q000295:instrumentation* / Q000592:standards; D006801:Humans; D015151:Immunoblotting / Q000592:standards; D008722:Methods; D009391:Nephelometry and Turbidimetry; D011697:Purpura, Thrombotic Thrombocytopenic / Q000175:diagnosis; D015203:Reproducibility of Results; D014841:von Willebrand Factor / Q000378:metabolism
D014841:von Willebrand Factor; D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human