冷冻干燥标本溶酶体酶稳定性的研究  

THE STABILITY OF FREEZE DRYING LYSOSOMAL ENZYMES

作　　者：郭玉凤[1] Boer,M 

机构地区：[1]中国医学科学院基础医学研究所

出　　处：《中华医学遗传学杂志》1998年第3期167-169,共3页Chinese Journal of Medical Genetics

摘　　要：目的为了解决溶酶体贮积症酶学诊断的标本在常温邮寄又不因外界条件的影响而导致酶的失活这一难题，探讨冷干技术处理标本后溶酶体酶的稳定性。方法建立正常人白细胞、血浆（各４０人份）和３株成纤维细胞标本库。对上述３个标本库分别进行了２０种、１４种和７种酶酶活性测定。比较了冷干后３种不同条件模拟邮寄与常规处理标本共４组酶活性的情况。结果经Ｆ检验：白细胞和成纤维细胞库４组之间酶活性无显著性差异（Ｐ＞０．０５）。血浆库４组之间酶活性有非常显著性差异（Ｐ＜０．０１）。经ｔ检验：白细胞和成纤维细胞库未冷干组与冷干后置室温３周组无显著性差异（Ｐ＞０．０５）。血浆库未冷干组与３周组有非常显著性差异（Ｐ＜０．０１）。Objective Enzyme assays for the diagnosis of lysosomal storage diseases have been underway in China only in a few laboratories,and the specimens for enzyme assay must not be inactivated by environmental factors.To solve the problem,the stability of freeze drying lysosomal enzymes was tested. Methods Three pools for samples were set up: control leucocytes pool and plasma pool(40 individuals in each pool),control fibroblasts pool(3 cell lines).4 groups of different dispositions of samples were designed for the pools:(1)pellets or plasma directly stored at -80℃ as routine (non freeze drying group or N); (2)after freeze drying, pellets or plasma immediately stored at -80℃(0 week or 0W);(3)after freeze drying,pellets or plasma stayed at room temperature for 1 week (simulated mail)then stored at -80℃(1 weeks or 1W);(4)the same with 1W but 3 weeks at room temperature(3 weeks or 3W). Microassay for enzyme activity was employed:20 enzymes in leucocytes pool,14 enzymes in fibroblasts pool and 7 in plasma pool. Results Enzyme activities among 4 groups in each sample pool were compared.Single factor analysis of variance( F test)showed no significance of differences( P >0 05) among 4 groups in leucocytes and fibroblasts pool,but highly significant differences ( P <0.01) among 4 groups in plasma pool.“ t test”showed no significance of differences( P >0.05) betweem N and 3W in leucocytes and also fibroblasts pool,but very significant differences ( P <0.01) between N and 3W in plasma pool. Conclusion The results suggest that freeze drying samples from leucocytes and fibroblasts can provide a stable resource for enzyme assay and be simple for transportation of the diagnostic samples.Though freeze drying is not appropriate for the plasma samples,it is still applicable for wide use since only a few kinds of lysosomal enzyme being indicated to use plasma for enzyme assay.

关 键 词：溶酶体贮积症 冷冻干燥 酶活性 测定 稳定性 

分 类 号：R589.9[医药卫生—内分泌]