全血标本采集管和保存温度与时间对HBV及HCV核酸检测的影响  

NAT results of HBV and HCV under different vacuum collection tubes,storage temperature and storage time

作　　者：张丽[1] 王艺芳[1] 吕永磊[1] 葛文超[1] 司天宁 朱晓红[1] 赵磊[1] ZHANG Li;WANG Yifang;LYU Yonglei;GE Wenchao;SI Tianning;ZHU Xiaohong;ZHAO Lei(Henan Red Cross Blood Center,Zhengzhou 450012,China)

机构地区：[1]河南省红十字血液中心,河南郑州450012

出　　处：《中国输血杂志》2021年第12期1314-1317,共4页Chinese Journal of Blood Transfusion

基　　金：河南省医学科技攻关项目(2018020586)。

摘　　要：目的验证不同厂家生产的核酸真空采血管和不同保存温度与时间对HBV及HCV核酸检测(NAT)结果的影响。方法选择2种(厂家)品牌的核酸真空采血管(分别标记为A和B组),做试验1:A和B组同时采集52名献血者的血液标本,比较不同采血管的血浆分离情况并分析不同保存时间对HBV DNA、HCV RNA的影响;试验2:模拟采血环境,验证不同保存温度、时间和2种采血管对A和B组低病毒载量HBV DNA、HCV RNA NAT结果的影响。结果试验1:1)离心后,A组标本血浆层和血细胞层均100%(52/52)分离,B组标本没有分离好血浆层和血细胞层的比例为1.92%(1/52);2)离心前保存4~10 h的2组(不同采血管)全血标本,离心后<24 h的NAT均未检出HBV DNA、HCV RNA,内标(IC-DNA和IC-RNA)均一性良好。试验2:1)离心前于4或25℃保存4 h离心或保存6~10 h离心,对2组中的低病毒载量HBV DNA、HCV RNA检出无影响(P>0.05);2)全血标本离心后分别在4℃保存24 h或72~104 h检测,A和B组标本HBV DNA、HCV RNA检测Ct值相近(P>0.05),保存时间24 h和72~104 h标本的HBV DNA、HCV RNA检测Ct值:A组HBV DNA Ct值为33.45±0.29和33.82±0.08,A组HCV RNA检测Ct值为35.21±0.20和36.12±0.43;B组为HBV DNA Ct值为33.46±0.25和34.30±0.60,B组HCV RNA检测Ct值为35.47±0.24和36.49±0.51(P<0.05)。结论在一定的实验室条件下,全血标本保存时间、温度及不同采血管对低病毒载量的HBV DNA、HCV RNA检出均无影响;但全血标本离心后4℃保存时完成检测时间以<72 h为宜。Objective To verify the results of HBV DNA and HCV RNA screening under different brands of vacuum collection tubes for blood samples,storage temperature and storage time.Methods Experiment 1 was conducted as follows:blood samples were collected simultaneously from 52 voluntary blood donors using two brands(divided into group A and group B)of vacuum collection tubes for blood samples.The plasma separation of group A and group B were compared,and the effects of storage time on the NAT yield of HBV DNA and HCV RNA were statistically analyzed.Experiment 2 was conducted as follows:the effects of different storage temperature,time and tubes on the NAT yield of HBV DNA and HCV RNA samples with low viral load in group A and B were verified and compared in the simulated phlebotomy condition.Results In Experiment 1:After centrifugation,blood plasma layer and cells layer were separated completely in group A(100%,52/52),but one sample was not well separated in group B(1/52,1.92%).After 4 to 10 h after collection,blood samples of two groups were centrifuged and screened for HBV DNA,HCV RNA within 24 h.No positive samples were yielded and the Ct values of internal control(IC-DNA and IC-RNA)were uniform.In Experiment 2:Whole blood samples,stored for either 4 h or 6~10 h at 4℃or 25℃before centrifugation,showed no difference on the NAT-yield of HBV DNA nor HCV RNA samples with low viral load(P>0.05).Ct values of HBV DNA and HCV RNA of group A was similar to those of group B as centrifuged samples were stored for 24 h or 72~104 h at 4℃(P>0.05),but all increased as the storage time prolonged.Ct values of HBV DNA in group A increased from 33.45±0.29(24 h)to 33.82±0.08(72~104 h)and HCV RNA from 35.21±0.20 to 36.12±0.43;HBV DNA from 33.46±0.25 to 34.30±0.60 and HCV RNA from 35.47±0.24 to 36.49±0.51 in group B.Conclusion Under certain laboratory condition,different storage time,storage temperature and tubes shed few effect on the NAT-yield of HBV DNA and HCV RNA samples with low virus loads.However,it is suggested that th

关 键 词：核酸检测 血液采集管 保存温度 保存时间 HBV DNA HCV RNA CT值 

分 类 号：R446.112[医药卫生—诊断学]