摘要
目的探讨不同温度和时间条件下保存对血液中丙型肝炎病毒核酸(HCV RNA)稳定性的影响。方法采集12名丙型肝炎患者4 ml的外周血,分装、保存在不同温度及相同温度下的不同时间,利用实时荧光定量PCR检测其不同时间点血浆HCV RNA的水平(log IU/ml),并与该血液标本的起始HCV RNA水平做比较分析。结果当血液标本中的HCV RNA起始水平≥105IU/ml时,在25℃放置6 h后转至4℃保存18 h(a)、25℃放置12 h后转至4℃保存12 h(b)及4℃放置24 h(c)等3种保存条件,24 h时内检测到血液标本的HCV RNA水平105IU/ml组分别为(5.15±0.16)、(5.15±0.15)及(5.16±0.16)IU/ml(P>0.05),107 IU/ml组分别为(7.45±0.21)、(7.44±0.23)及(7.45±0.24)IU/ml(P>0.05);当血液标本中HCV RNA起始水平在临界范围[(3.38±0.14)IU/ml]时,在条件a、b保存24 h血液标本的HCV RNA含量分别为(2.28±1.52)IU/ml、0(均为P<0.01),而条件c保存24 h血液标本的HCV RNA含量为(3.28±0.20)IU/ml(P>0.05);当血液标本中HCV RNA起始水平为103 IU/ml时,在4℃保存48 h时HCV RNA含量为(2.31±1.54)IU/ml(P<0.01)。结论当血液中HCVRNA起始浓度较低时,HCV RNA的稳定性较差,短时常温保存都会造成病毒核酸的降解。
Objective To investigate the influence of preservation temperature and time of blood sample on the stabilityof hepatitis c virus RNA （HCV RNA）. Methods 12 patients with hepatitis C were included in this study, and 4 ml peripheral blood was sampled from each one. Blood sample were distributed and preserved in different temperature for different time. Then,RNA was extracted and real-time fluorescent quantitative PCR was performed to test the plasma HCV RNA level at different time point, and was compared with the inceptive HCV RNA concentration. Results As the inceptive HCV RNA level is/〉 105 IU/ml in the blood sample,the HCV RNA level was no significant variation after preserving in the 3 conservation condi- tions （a. 25℃ for 6 h then transfer to 4℃ for 18 h;b. 25℃ for 12 h then transfer to 4℃ for 12 h;c. 4℃ for 24 h） for 24 hours [（5.15 ±0. 16） ,（5.15 ±0. 15） and（5. 16 ±0. 16） IU/ml,respectively in 10^5 IU/ml group,and （7.45 ±0.21）, （7.44 ± 0. 23 ）, and （ 7. 45 ± 0. 24 ） IU/ml, respectively, in 107 IU/ml group ] compared with the inceptive concentration （ P 〉 0. 05 ） . However,when the inceptive HCV RNA level is 103 IU/ml in the blood sample,HCV RNA level significantly decreased after preserving in the condition a and b for 24 h [ （2. 28 ± 1.52） ,0 IU/ml,respectively] （P 〈0. 01 ） ,and was not affected in the conservation condition c[ （3.28 ±0. 20） IU/nd] （P 〉0. 05） ;HCV RNA was found to be degraded after preservation for 6 h in 25℃ ,and for 48 h in 4℃ [ （2. 31 ± 1.54） IU/ml], when the inceptive HCV level is 103 IU/ml. Conclusion Among the low concentration of blood simple and room temperature ,HCV RNA would be degenerated in short time.
出处
《中国输血杂志》
CAS
CSCD
北大核心
2012年第6期549-552,共4页
Chinese Journal of Blood Transfusion
基金
广东省优秀博士论文作者资助项目(sybzzxm201127)
关键词
丙型肝炎病毒
RNA
保存条件
温度
时间
荧光定量PCR
Hepatitis e virus
RNA
Preservation condition
Temperature
Time
Fluorescent quantitative PCR
