冻存温度、冻融次数和冻存时间对血清生化指标稳定性的影响  

Effects of temperature,freeze-thaw cycles and storage time on stability of serum biomarkers

作　　者：范泽涵 章月蟾 索晨 徐珂琳 薛江莉 李淑元[5] 袁子宇 王久存[2] 金力[2] 陈兴栋[2,4] FAN Zehan 1, ZHANG Yuechan 1, SUO Chen 2, XU Kelin 3, XUE Jiangli 4,LI Shuyuan 5, Yuan Ziyu 4, WANG Jiucun 2, JIN Li 2, CHEN Xingdong 2,4(1.State Key Laboratory of Genetic Engineering and MOE Key Laboratory of Contemporary Anthropology,School of Life Sciences, Fudan University, Shanghai 200438, China; 2.The State Key Laboratory of GeneticEngineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, Fudan University,Shanghai 200438, China; 3.School of Data Science, Collaborative Innovation Center for Genetics and Development,Fudan University, Shanghai 200438, China; 4.Fudan University Taizhou Institute of Health Sciences, Taizhou Jiangsu225300, China; 5.Institute of Embryo-Fetal Original Adult Disease, International Peace Maternity andChild Health Hospital, University School of Medicine, Shanghai Jiaotong, Shanghai 200030, Chin)

机构地区：[1]复旦大学生命科学学院现代人类学教育部重点实验室,上海200438 [2]复旦大学遗传工程国家重点实验室、遗传与发育协同创新中心,上海200438 [3]复旦大学数据学院、遗传与发育协同创新中心,上海200438 [4]复旦大学泰州健康科学研究院,江苏泰州225300 [5]上海交通大学医学院附属国际和平妇幼保健院生殖遗传科、上海交通大学医学院胚胎源性疾病研究所,上海200030

出　　处：《转化医学杂志》2018年第4期212-217,共6页Translational Medicine Journal

基　　金：国家自然科学基金(8150120276);中国博士后基金(2016M601511、2017T100272);国家重点研发计划(2017YFC0907002)

摘　　要：目的分析在生物样本库中,冻存温度、冻存时间及反复冻融效应对血清常规生化指标的影响,以探讨血清常规生化指标在生物样本库中的稳定性。方法以2014年7月江苏省泰兴市人民医院160例健康体检者为研究对象,取空腹静脉血制备混合血清。24 h内检测新鲜混合血清的12项常规生化指标作为基准值。将混合血清样本随机分组:A组贮存于-20℃,B组、C组、D组、E组均贮存于-80℃,其中C组、D组、E组分别于冻存开始的1、5、10 d内,每天进行一次冻融处理。在第1,3,5,7,9个月时进行重复检测。运用配对t检验比较各时点检测值与基准值的差异,运用线性混合效应模型分析冻存温度、冻融循环次数、冻存时间效应对各指标含量稳定性的影响。结果 9个月的冻存中,12项生化指标的含量均不稳定。导致含量不稳定的因素有冻存温度、反复冻融次数、冻存时间,冻存温度、冻融次数与冻存时间分别产生的交互效应。结论血清样本不宜在生物样本库中冻存,应选择新鲜样本进行血清常规生化指标的检测,否则会降低检测结果的准确性。Objective This study aimed at investigating the effects of temperature,freezethaw cycles and storage time on the stability of common clinical serum biomarkers in the biobank.Methods Serum samples were collected from 160 healthy adults. They were tested for target biomarkers within 24 hours and randomly divided into five groups（ group A to E）. Group A was stored at-20 ℃ and group B to E at-80 ℃. Group C to E had one,five and ten repeated freeze-thaw cycles respectively during the first few days of storage. Then we tested them after one,three,five,seven,nine months and compared the value with baseline. Changes in mean component concentrations were evaluated using paired t-tests. The linear mixed effect model was used to investigate the effect of time,temperature,freeze-thaw cycles and their interactions on the stability of the biomarkers. Results During nine months,all target biomarkers showed unstable fluctuations in each group. Temperature,freeze-thaw cycles,the interactions between storage time and temperature,the interactions between storage time and freeze-thaw cycles would influence the stability of serum biomarkers. Conclusion Biobanking would reduce the validity and stability of serum samples,thus serum biomarkers should be tested as soon as possible.

关 键 词：生物样本库 血清 生化指标 温度 反复冻融 

分 类 号：R446.1[医药卫生—诊断学]