Department of Infectious Diseases, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, 1# Shuai Fu Yuan, Beijing, 100730, China.
Center for AIDS Research, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
BMC Infect Dis. 2020 Dec 10;20(1):941. doi: 10.1186/s12879-020-05675-3.
A more time saving, convenient, reproducible, and scalable method is needed to assess total HIV-1 DNA levels.
Frozen whole blood and peripheral blood mononuclear cell (PBMC) samples both 200 μl at the same point were used to detect total HIV-1 DNA. Automatic extraction of total HIV-1 DNA was used to ensure the consistency of sample extraction efficiency. The detection reagent was HIV-1 DNA quantitative detection kit and real-time quantitative PCR was utilized.
Of the 44 included patients, 42 were male and 2 were female, with a median age of 33 years. Thirty-three cases were collected after receiving antiviral treatment, with a median duration of treatment of 3 months, and the other 11 cases were collected before antiviral treatment. The median viral load was 1.83 log10 copies/mL, the median CD4 and CD8 count were 94 and 680 cells/μL, and the median CD4/CD8 ratio was 0.18. The results of the two samples were 3.02 ± 0.39 log10 copies/10 PBMCs in PBMC samples and 3.05 ± 0.40 log10 copies/10 PBMCs in whole blood samples. The detection results of the two methods were highly correlated and consistent by using paired t test (P = 0.370), pearson correlation (r = 0.887, P < 0.0001) and intra-group correlation coefficient (ICC = 0.887, P < 0.0001) and bland-altman [4.55% points were outside the 95% limits of agreement (- 0.340 ~ 0.390)].
The results of the whole blood sample test for total HIV-1 DNA are consistent with those of PBMC samples. In a clinical setting it is recommended to use whole blood samples directly for the evaluation of the HIV reservoir.
需要一种更省时、方便、可重现和可扩展的方法来评估总 HIV-1 DNA 水平。
使用相同量的 200μl 冷冻全血和外周血单核细胞（PBMC）样本来检测总 HIV-1 DNA。使用自动提取总 HIV-1 DNA 来确保样本提取效率的一致性。检测试剂为 HIV-1 DNA 定量检测试剂盒，采用实时定量 PCR 法。
纳入的 44 例患者中，男 42 例，女 2 例，中位年龄 33 岁。33 例为接受抗病毒治疗后采集的标本，中位治疗时间 3 个月，11 例为抗病毒治疗前采集的标本。中位病毒载量为 1.83log10 拷贝/ml，中位 CD4 和 CD8 计数分别为 94 和 680 个/μl，中位 CD4/CD8 比值为 0.18。PBMC 样本中 PBMC 样本的检测结果为 3.02±0.39log10 拷贝/10 PBMC，全血样本中为 3.05±0.40log10 拷贝/10 PBMC。配对 t 检验（P=0.370）、pearson 相关系数（r=0.887，P<0.0001）和组内相关系数（ICC=0.887，P<0.0001）均显示两种方法的检测结果高度相关且一致， Bland-Altman 分析[4.55%的点超出 95%一致性界限（-0.340~0.390）]。
全血样本总 HIV-1 DNA 的检测结果与 PBMC 样本一致。在临床环境中，建议直接使用全血样本评估 HIV 储存库。
