【摘要】 目的测定机采血小板(AP-PCs)中游离DNA的含量,并分析其影响因素。方法从53例合格献血者各采集AP-PCs1个治疗剂量后,尽快从其中留取标本,并将其分成5份。其中1份用于血液分析仪检测,另外4份于采集后4h内、24、48和72h时,分别在过滤前后、离心前后等不同状态下进行DNA抽提,用实时定量PCR方法(RQ-PCR)对其中的人类白蛋白基因进行检测,并将结果换算成白细胞当量(WBCs/μl)。结果AP-PCs在采集后的4h内、24、48和72h时,其过滤前游离DNA的含量(WBCs/μl)分别为13.3±6.1,24.7±10.7,47.5±20.1和146.3±63.2;过滤后其相应值(WBCs/μl)则为8.1±3.7,22.2±9.4,44.7±18.7和125.8±57.9。过滤前后游离DNA含量差异无统计学意义(F=2.922,P>0.05),不同保存时间组之间游离DNA含量差异具有统计学意义(F=512.734,P<0.01)。ABO血型不同的AP-PCs经过相同保存时间后,其过滤前游离DNA含量差异无统计学意义(F=0.662,P>0.05)。结论AP-PCs中游离DNA的含量随着保存时间的延长而上升,但与过滤与否及供者血型无关。更多还原

【Abstract】 Objective To detect the extent of cell-free DNA in apheresis platelet concentrates(AP-PCs),and to find out influences of storage time,filtration and ABO blood group on the concentration of it.Methods 53 therapeutic dose of AP-PCs were sampled after donation as soon as possible.Then the samples were divided into 5 parts.One part was analyzed by hematology analyzer,the others underwent 4 DNA extractions in different conditions:before or after filtration,and before or after centrifugation at the time of no more than 4 hours,24 hours,48 hours and 72 houres after donation respectively.Then the amount of total DNA of the AP-PCs and the cell-free DNA in plasma was determined using real-time quantitative-polymerase chain reaction(RQ-PCR) for the albumin gene,and the results were calculated into leukocytes equivalent.Results The amount of cell free DNA(WBCs/μl) in unfiltered AP-PCs at <4 hours,24 hours,48 hours and 72 houres after apheresis were 13.3±6.1,24.7±10.7,47.5±20.1 and 146.3±63.2 respectively.After filtration,the corresponding values(WBCs/μl) were 8.1±3.7,22.2±9.4,44.7±18.7 and 125.8±57.9.There was no significant difference between the groups of before and after filtration(F=2.922,P>0.05),but there were significant differences among the groups of different storage time(F=512.734,P<0.01).There was no significant difference among the concentration of cell-free DNA in AP-PCs of different ABO blood groups(F=0.662,P>0.05).Conclusions The amount of cell-free DNA arises sharply during storage,but it isn’t influenced by filtration and ABO blood group.更多还原