Simple method for α1-antitrypsin deficiency screening by use of dried blood spot specimens
Simple method for α1-antitrypsin deficiency screening by use of dried blood spot specimens
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DOI：
10.1034/j.1399-3003.2000.01521.x
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发表时间：
2000-06-01
影响因子：
24.3
通讯作者：
Vidal, R
中科院分区：
文献类型：
--
作者：
Costa, X;Jardi, R;Vidal, R
干血斑 (DBS) 样本在定量 α(1)-抗胰蛋白酶 (α(1)-AT) 检测或遗传分析中的使用受到限制，因为样本中的蛋白质水平较低，并且含有可能干扰聚合酶链式反应扩增的成分。我们开发了一种方法来克服本文讨论的这些缺点。研究人群包括 200 名健康志愿者和 300 名慢性阻塞性肺病 (COPD) 患者，使用改良的浊度测定法测试 DBS 样本的 α(1)-AT 浓度，并使用等电聚焦方法进行表型分析。通过脱氧核糖核酸测序建立遗传诊断，使用简单的纯化程序去除污染物。比浊法显示检测限为0.284 mg.dL(-1)，对应于血清浓度13 mg.dL(-1)。 DBS 与血清样本中的 α(1)-AT 浓度之间的相关系数为 R-2=0.8674 (p1.9 mg.dL，对应于血清样本中的 114 mg.dL(-1)。125 名 COPD 患者 (42%) 显示 α(1)-AT 值
The use of dried blood spot (DBS) specimens in quantitative alpha(1)-antitrypsin (alpha(1)-AT) detection or genetic analysis is limited because protein levels in the samples are low and they contain components that can interfere with polymerase chain reaction amplification. A methodological adaptation was developed to overcome these drawbacks which is discussed here.The study population consisted of 200 healthy volunteers and 300 patients with chronic obstructive pulmonary disease (COPD), DBS specimens were tested for alpha(1)-AT concentration using a modified nephelometric assay and phenotyped with an isoelectric focusing method. Genetic diagnosis was established by deoxyribonucleic acid sequencing using a simple purification procedure to remove contaminants.The nephelometric method showed a detection limit of 0.284 mg.dL(-1), corresponding to a serum concentration of 13 mg.dL(-1). The correlation coefficient between alpha(1)-AT concentrations in DBS versus serum samples was R-2=0.8674 (p1.9 mg.dL, corresponding to 114 mg.dL(-1) in serum samples. One hundred and twenty-five COPD patients (42%) showed alpha(1)-AT values