Crist R A, Heikal N M, Rodgers G M, Grenache D G, Smock K J
ARUP Laboratories Institute for Clinical and Experimental Pathology, University of Utah, Salt Lake City, UT, USA.
Department of Pathology, University of Utah, Salt Lake City, UT, USA.
Int J Lab Hematol. 2018 Oct;40(5):586-591. doi: 10.1111/ijlh.12869. Epub 2018 Jun 19.
Evaluation of von Willebrand factor (VWF) multimeric distribution is useful for subclassification of von Willebrand disease (VWD). Multimer analysis has historically been a manual, labor-intensive laboratory-developed test. The first commercial method for multimeric analysis was recently developed that utilizes a single instrument for gel electrophoresis, staining, and densitometry. The current study was undertaken to evaluate the performance characteristics of the new commercial method.
Studies performed with the commercial method included evaluation of accuracy (method comparison), reference intervals (establishment of normal migration patterns in normal donor specimens), precision (multimer pattern reproducibility), and analytical sensitivity.
In the method comparison studies, concordant interpretations were obtained in 19 of 24 comparisons, including normal and abnormal specimens. The 5 specimens with discordant interpretations all involved slight differences and none were considered clinically significant. Thirty-eight normal donor specimens demonstrated normal multimer patterns. Multimer pattern reproducibility was demonstrated in normal and abnormal controls tested on each gel. In the sensitivity studies, adequate visualization of multimers was determined to require VWF protein concentrations of approximately 5%-10% of normal.
The commercial multimer method is a streamlined test that demonstrates comparable performance characteristics to our current laboratory-developed method and that provides the advantage of both electrophoresis gels and densitometry scans to aid interpretation.
评估血管性血友病因子(VWF)多聚体分布对于血管性血友病(VWD)的亚分类很有用。多聚体分析历来是一种手工操作、劳动强度大的实验室自行开发的检测方法。最近开发了第一种用于多聚体分析的商业方法,该方法使用单一仪器进行凝胶电泳、染色和光密度测定。本研究旨在评估这种新商业方法的性能特征。
使用商业方法进行的研究包括准确性评估(方法比较)、参考区间(确定正常供体标本中的正常迁移模式)、精密度(多聚体模式的可重复性)和分析灵敏度。
在方法比较研究中,24次比较中的19次获得了一致的解释,包括正常和异常标本。5份解释不一致的标本均存在细微差异,且均不被认为具有临床意义。38份正常供体标本显示出正常的多聚体模式。在每个凝胶上测试的正常和异常对照中均证明了多聚体模式的可重复性。在灵敏度研究中,确定多聚体的充分可视化需要VWF蛋白浓度约为正常浓度的5%-10%。
商业多聚体方法是一种简化的检测方法,其性能特征与我们目前实验室自行开发的方法相当,并且具有电泳凝胶和光密度测定扫描两者的优势,有助于解释结果。