溶血、脂肪血及保存条件对HBV DNA、HIV RNA核酸检测的影响

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.%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.