一种用于³H-DNA和血清脱氧核糖核酸酶的改良滤膜圆盘测定法。

An improved filter-disc assay for 3H-DNA and serum deoxyribonucleases.

Cox R A, Gokcen M

Clin Biochem. 1977 Oct;10(5):188-90. doi: 10.1016/s0009-9120(77)92842-9.

Abstract

The filter-paper disc assay for tritium-labeled nucleic acids was modified to reduce washing and manipulation. Evaluation of the procedure indicated greatly enhanced counting efficiency for tritium, complete removal of nuclease digestion products and retention of even nanogram levels of radioactive DNA on the discs in the presence of protein. The modified assay was quite sensitive and had a coefficient of variation of 5.2%. Quite low concentrations of DNase I and endogenous human serum nuclease activity were readily measured by the improved technique, 22 sera yielding a mean value equivalent to about 0.5 ng DNase I/ml serum. We are now using the disc assay to follow serum DNA and DNase levels in patients with autoimmune disorders and various types of cancers.

摘要

对用于氚标记核酸的滤纸圆盘测定法进行了改进，以减少洗涤和操作。对该方法的评估表明，氚的计数效率大大提高，核酸酶消化产物被完全去除，并且在有蛋白质存在的情况下，圆盘上仍能保留纳克级的放射性DNA。改进后的测定法相当灵敏，变异系数为5.2%。通过改进技术能够轻松测定相当低浓度的DNase I和内源性人血清核酸酶活性，22份血清得出的平均值相当于约0.5 ng DNase I/ml血清。我们现在正在使用圆盘测定法来跟踪自身免疫性疾病和各种癌症患者的血清DNA和DNase水平。