A simple method for quantification of biotinidase activity in dried blood spot and its application to screening of biotinidase deficiency.
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.
Yamaguchi
A
A
Sapporo City Institute of Public Health, Japan.
Fukushi
M
M
Arai
O
O
Mizushima
Y
Y
Sato
Y
Y
Shimizu
Y
Y
Tomidokoro
K
K
Takasugi
N
N
D000328:Adult; D000375:Aging / Q000097:blood; D000581:Amidohydrolases / Q000097:blood* / Q000172:deficiency; D043523:Biotinidase; D001800:Blood Specimen Collection / Q000379:methods; D003124:Colorimetry / Q000379:methods; D006454:Hemoglobins; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007234:Infant, Premature; D008403:Mass Screening / Q000379:methods*; D011489:Protein Denaturation
D006454:Hemoglobins; D000581:Amidohydrolases; D043523:Biotinidase