一种用于定量干血斑中生物素酶活性的简单方法及其在生物素酶缺乏症筛查中的应用。

A simple method for quantification of biotinidase activity in dried blood spot and its application to screening of biotinidase deficiency.

作者信息

Yamaguchi A, Fukushi M, Arai O, Mizushima Y, Sato Y, Shimizu Y, Tomidokoro K, Takasugi N

机构信息

Sapporo City Institute of Public Health, Japan.

出版信息

Tohoku J Exp Med. 1987 Aug;152(4):339-46. doi: 10.1620/tjem.152.339.

DOI:10.1620/tjem.152.339

PMID:3660404

Abstract

A simple and reliable method for quantification of biotinidase (EC.3.5.1.12) activity in dried blood spot was devised by a modification of the colorimetric screening test developed by Heard et al. (1984). The enzyme reaction and hemoglobin denaturation were carried out in a U-bottomed microplate. An aliquot of the reaction solution was transferred to a flat-bottomed microplate. After the coupling reaction was started, the adsorbance was measured in situ by a microplate-reader. Both intra- and inter-assay coefficient of variation (CV) values were less than 10%. Biotinidase activity in dried blood spot showed a good correlation to that in serum (r = 0.912, n = 8). This method was applied in a pilot screening of 18,945 newborns in Sapporo City. No positive results have been obtained as yet.

摘要

通过对Heard等人（1984年）开发的比色筛选试验进行改进，设计出了一种简单可靠的方法，用于定量干血斑中的生物素酶（EC.3.5.1.12）活性。酶反应和血红蛋白变性在U型底微孔板中进行。将一份反应溶液转移至平底微孔板中。偶联反应开始后，用微孔板读数仪原位测量吸光度。批内和批间变异系数（CV）值均小于10%。干血斑中的生物素酶活性与血清中的生物素酶活性具有良好的相关性（r = 0.912，n = 8）。该方法应用于对札幌市18945名新生儿的初步筛查。目前尚未获得阳性结果。