自然沉降法分离抗凝全血建立生物样本库的特点  

Characteristic of sample banks isolated from EDTA-blood by sedimentation method

作　　者：陈智滨[1] 林琴[2] 马长华[2] 刘凯宁[1] 孟焕新[1] 

机构地区：[1]北京大学口腔医学院.口腔医院牙周科,北京100081 [2]北京大学口腔医学院.口腔医院检验科,北京100081

出　　处：《北京大学学报（医学版）》2014年第1期111-114,共4页Journal of Peking University:Health Sciences

基　　金：国家临床重点专科建设项目;"十一五"国家科技支撑计划(2007BAI18B02);卫生部部属(管)医院临床学科重点项目;北京大学口腔医学院新技术;新疗法基金资助~~

摘　　要：目的：评估用自然沉降法分离EDTA抗凝外周静脉全血（抗凝全血）建立血浆、白细胞、基因组DNA生物样本库的特点.方法：采集符合纳入标准的29名志愿者抗凝全血,进行血常规检测作为对照组;然后将抗凝全血分装到4支EP管中,每支1 mL,分为0.5、1、2、3 h 4个试验组,于室温静置自然沉降0.5、1、2、3 h后吸取血浆层,测量其体积,检测血浆层中白细胞总数及中性粒细胞、淋巴细胞的百分比;将含有白细胞的血浆层离心后,分装为无细胞血浆管和白细胞沉淀管,用部分白细胞提取DNA,建立血浆、白细胞、DNA 3种生物样本库.结果：0.5 h组血浆体积（241.72±101.52）μL显著低于1 h组（317.24±97.50）μL、2 h组（371.03±91.66）μL和3 h组（408.97±97.43）μL,P〈0.05.1 h组血浆体积显著低于2 h组和3 h组,P〈0.05;2 h组显著低于3 h组,P〈0.05.0.5 h组白细胞总数[（2.50×106）±（1.48×106）]显著高于2 h组[（1.47×106）±（7.19×105） ]和3 h组[（1.21×106）±（7.41×105）]的白细胞总数,P〈0.05,与1 h组[（2.29×106）±（1.17×106）]比较差异无统计学意义,P〉0.05;1 h组显著高于2 h和3 h组,P〈0.05;2 h组与3 h组差异无统计学意义,P〉0.05.4个试验组白细胞总数均显著低于对照组.中性粒细胞百分比：0.5 h组（54.14%±11.65%）显著高于1、2、3 h组（46.66%±12.70%,39.17%±12.33%,43.25%±14.54%）,P〈0.05,显著低于对照组（60.53%±8.46%）,P〈0.05;1 h组显著高于2 h组,P〉0.05; 1 h组与3 h组比较差异无统计学意义,P〉0.05;3 h组与1 h、2 h组比较差异无统计学意义,P〉0.05.淋巴细胞百分比：0.5 h组（35.09%±10.84%）显著低于1、2、3 h组（41.48%±12.20%、47.96%±12.27%、45.50%±13.71%）,显著高于对照组（30.98%±7.33%）,P〈0.05;1 h组显著高于对照组、0.5 h组,显著低于2 h、3 h组,P〈0.05;2 h组显著高于对照组、0.5 h、1 h组,P〈0.05,与3 h组之间差异无统计学意义,P〉0.05.结论：抗凝全血Objective：To assess the characteristics of establishing the different sample banks of plas- ma, leukocytes and DNA by sedimentation method of isolating from ethylene diamine tetraacetic acid （EDTA） -blood and to clarify the sedimentation method of leukocyte isolation and plasma volume by com- parative data and recommended procedures for applicability. Methods：In the study, 29 EDTA-bloods were obtained, the total amounts of leukocytes and the percentage of neutrophile granulocytes, and lym- phocytes in the EDTA-blood detected as a control group and then assigned equally into 4 EP tubes with 1 mL EDTA-hlood per tube as 4 test groups, then the 4 tubes were placed with the EDTA-blood at room temperature and the plasma layers were isolated at 0.5, 1, 2 and 3 h, receptively. The total amount of leukocytes and the percentage of neutrophile granulocytes, and lymphocytes were detected by automated hematology analyzer at the clinical laboratory. The volume of the plasma was also measured at the same time. Results：The plasma volume at 0.5 h [ （241.72 ± 101.52） μL] was substantially lower than those at 1 hi （317.24 ±97.50）μL], at 2 hi （371.03 ±91.66）μL] , and at 3 h [ （408.97±97.43） μL] , P 〈 0.05. The plasma volume at 1 h was substantially lower than those at 2 h and 3 h （P 〈 0.05 ）. The total amount of leukocytes in the plasma layer at 0.5 h （2.50 × 106 ± 1.48× 106） group was substantially higher than the amount of 2 or 3 h groups respectively （ 1.47 × 106 ± 7.19 ×106 , 1.21×106- 7. 41× 105 ）, P 〈0.05. Significant difference was not found between 0.5 h group and 1 h group （2.29×106 ± 1. 17× 106） , P 〉0.05. The total amount of leukocytes in the plasma layer inlh group was substantially higher than that in 2 h and 3 h groups （ P 〈 0.05 ）. There was no significant difference between 3 h group and 2 h group （P 〉 0.05 ）. The total amount of leukocytes in the plasma layer of the 4 test groups was substantially lower than that in the control group �

关 键 词：血沉 生物标本库 血浆 白细胞计数 DNA 

分 类 号：R331.14[医药卫生—人体生理学]