天津医药

天津医药 ›› 2017, Vol. 45 ›› Issue (10): 1025-1028.doi: 10.11958/20170524

• 实验研究 • 上一篇    下一篇

溶血、脂肪血及保存条件对 HBV DNA、HIV RNA 核酸检测的影响

庄养林 1，熊丽红 1，张鹏飞 1，王芳 1，陈丹 2   

  1. 1 江西省血液中心检验科（邮编 330052）；2 江西省职业病防治研究院
  • 收稿日期:2017-04-28 修回日期:2017-06-29 出版日期:2017-10-15 发布日期:2017-10-13
  • 通讯作者: 庄养林 E-mail:290664367@qq.com
  • 作者简介:庄养林（1985），男，硕士，主管技师，主要从事输血检验研究
  • 基金资助:
    江西省卫生计生委科技计划

Study on the effects of hemolysis, fatty blood and various storage conditions on HBV DNA, HIV RNA nucleic acid screening

ZHUANG Yang-lin1, XIONG Li-hong1, ZHANG Peng-fei1, WANG Fang1, CHEN Dan2   

  1. 1 Clinical Laboratory of Jiangxi Blood Center, Nanchang 330052, China; 2 Institute of Occupational Medicine of Jiangxi
  • Received:2017-04-28 Revised:2017-06-29 Published:2017-10-15 Online:2017-10-13

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摘要： 目的 研究溶血、脂肪血、不同保存温度和保存时间对血液 HIV RNA、HBV DNA 核酸检测（NAT）的影 响。方法 应用罗氏 MPX V2.0 试剂，分别于 4 ℃、25 ℃、37 ℃及-30 ℃下进行保存的 12~30 倍试剂检测下限（LOD） 浓度的 HIV RNA、HBV DNA 标本，对所有标本于 4 h、1 d、2 d、3 d、1 周及 4 周后，采用 6 混样模式进行 NAT 检测；对 含终浓度为 2~5 倍 LOD 的 HIV RNA、HBV DNA 标本的 6 组溶血标准样本［血红蛋白（Hb）浓度分别为 97、34、17、8、 5 及 3 g/L］、5 组脂肪血标准［三酰甘油（TG）浓度分别为 7.93、3.80、2.63、1.83 及 1.49 mmol/L］、1 组无溶血正常三酰 甘油浓度（Hb 及 TG 浓度分别为 0 g/L、0.95 mmol/L）的对照样本进行 NAT 检测。结果 在 25 ℃及以下温度保存 4 h 至 4 周，HIV RNA、HBV DNA 检测循环阈值（Ct 值）差异均无统计学意义（P＞0.05），HBV DNA 在 37 ℃条件下保存， 3 d 及 1 周检测 Ct 值分别与 4 h、1 d、2 d 比较差异有统计学意义（P＜0.05）；Hb 浓度为 97 g/L 时 HBV DNA 和 HIV RNA 均未能被检出，当 Hb 浓度＜34 g/L 时，HBV DNA、HIV RNA 检测 Ct 值分别与其对照组相比较差异无统计学意 义（P＞0.05）；TG≤7.93 mmol/L 时，HIV RNA、HBV DNA 各组检测 Ct 值与对照组比较差异无统计学意义（P＞0.05）。 结论 使用罗氏 MPX V2.0 试剂检测的 HIV RNA、HBV DNA 标本，在 25 ℃及以下最长可以放置 4 周；Hb＜34 g/L 以 及 TG≤7.93 mmol/L 时，对罗氏 MPX V2.0 试剂检测 HIV RNA、HBV DNA 没有显著性影响。

关键词: 血液保存, 因素分析, 统计学, 乙肝病毒脱氧核糖核酸, 人类免疫缺陷病毒核糖核酸, 核酸检测技术, Ct 值, 溶血, 脂肪血

Abstract: Objective To study the influences of hemolysis, fatty blood, storage temperature and storage time on blood HIV RNA and HBV DNA. Methods The HBV DNA and HIV RNA samples (the concentration was 12-30 times of limit of detection of reagent), which were stored for 4 h, 1 d, 3 d,1 w and 4 w under the conditions of 4 ℃, 25 ℃, 37 ℃ and -30 ℃, were detected using Roche MPX V2.0 kit. The HBV DNA and HIV RNA samples (the concentration was 2-5 times of limit of detection of reagent) were detected by the 6 groups of hemolytic samples (the concentrations of hemoglobin were 97 g/L, 34 g/L,17 g/L, 8 g/L, 5 g/L and 3 g/L, respectively). NAT test was performed at 5-group lipid samples (the concentrations of triglyceride were 7.93 mmol/L, 3.80 mmol/L, 2.63 mmol/L, 1.83 mmol/L and 1.49 mmol/L, respectively) and the control group samples (the concentrations of hemoglobin and triglyceride were 0 g/L and 0.95 mmol/L respectively). Results There were no significant differences in Ct values of HIV RNA or HBV DNA between 4 h to 4 w at 25 ℃ (P＞0.05). There were significant differences in Ct values of HBV DNA after preservation for 3 d and 1 w under 37 ℃ compared with those of preservation for 4 h,1 d and 2 d (P＜0.005). When Hb concentration was reached to 97 g/L, the results of HBV DNA and HIV RNA were negative. When the concentration of Hb was less than 34 g/L, compared with the control group, there were no significant differences in Ct values of HIV RNA and HBV DNA (P＞0.05). When the TG concentration was ≤ 7.93 mmol/L, there were no significant differences in Ct values between the control group and the TG group (P＞0.05). Conclusion The samples of HIV RNA and HBV DNA detected by Roche MPX V2.0 kit can be stored at room temperature (25 ℃) for four weeks. When the concentrations of TG and Hb are less than 7.93 mmol/L and 34 g/L respectively, there are no effects on HIV RNA or HBV DNA samples detected by Roche MPX V2.0 kit.

Key words: blood preservation, factor analysis, statistical, hepatitis B virus deoxyribonucleic acid, human immunodeficiency virus ribonucleic acid, nucleic acid testing；cycle threshold value；hemolysis, fatty blood