Maeda K, Ito K, Yamaguchi N
Clin Chim Acta. 1980 Jan 15;100(2):175-81. doi: 10.1016/0009-8981(80)90080-7.
A simple method for quantifying lysozyme in serum and urine has been established by modifying the procedures of sample application in the lysoplate method reported by Osserman and Lawlor [1]. A strip of filter paper is partially immersed in a liquid sample, dried at room temperature and cut into discs which are later placed on the surface of an agarose gel containing Micrococcus lysodeikticus. The follow-ng procedures for determination are carried out as described in the lysoplate method. There was no statistically significant loss of enzyme activity during the sample preparation. Lysozyme dried on filter paper is so stable that it can be stored at room temperature for at least 13 weeks and can be mailed. The sensitivity of the method is increased by pretreatment of the filter paper with Triton X-100 and consequently corresponds to that of the lysoplate method. The reproducibility of our method is practically good since the variation coefficient of the diameters of lytic zones within assays is around 1%. There is a very close correlation between lysozyme levels determined by this method and by the lysoplate method with serum samples obtained from patients with various hematological diseases (r = +0.994). The method can be utilized for routine clinical determinations of lysozyme.
通过改进奥瑟曼和劳勒[1]报道的溶菌平板法中的样品施加步骤,建立了一种简单的血清和尿液中溶菌酶定量方法。将一条滤纸部分浸入液体样品中,在室温下干燥,然后切成圆片,随后将这些圆片放置在含有溶壁微球菌的琼脂糖凝胶表面。测定的后续步骤按照溶菌平板法中的描述进行。在样品制备过程中,酶活性没有统计学上的显著损失。滤纸上干燥的溶菌酶非常稳定,可在室温下保存至少13周且可邮寄。通过用曲拉通X-100预处理滤纸,该方法的灵敏度得以提高,因此与溶菌平板法相当。由于测定内溶菌圈直径的变异系数约为1%,我们方法的重现性实际上很好。用该方法测定的溶菌酶水平与用溶菌平板法测定的来自各种血液系统疾病患者的血清样品中的溶菌酶水平之间存在非常密切的相关性(r = +0.994)。该方法可用于溶菌酶的常规临床测定。