干血斑用于HIV-1新发感染检测(限制性抗原亲和力法)研究

Dry blood spots used in the detection of HIV-1 recent infection(limiting antigen avidity enzyme immunoassay, Lag-Avidity EIA)

目的探讨干血斑样本用于1型艾滋病病毒(HIV-1)新发感染检测(限制性抗原亲和力法)的可行性与最佳检测条件。方法用制备的三套系列稀释干血斑-血浆配对样本和来自临床的71对人群干血斑-血浆配对样本,使用国产HIV-1新发感染酶免检测试剂盒(限制性抗原亲和力法),同时对血浆样本与干血斑样本进行检测并分析等效性。其中滤纸片干血斑(DBS)样本(6mm斑)洗脱分别在500μL、600μL、800μL和1 000μL样本稀释液4℃过夜洗脱。结果三套系列稀释干血斑-血浆配对样本检测结果显示,500μL、600μL、800μL和1 000μL样本稀释液洗脱后的DBS样本与血浆样本的Pearson相关系数分别依次为0.955(R^2=0.912)、0.944(R^2=0.892)、0.948(R^2=0.899)、0.933(R^2=0.870);来自临床的71对人群干血斑-血浆配对样本检测结果显示,500μL、600μL、800μL和1 000μL样本稀释液洗脱后的DBS样本与血浆样本的Pearson相关系数分别依次为0.968(R^2=0.936)、0.965(R^2=0.932)、0.959(R^2=0.919)、0.879(R^2=0.773);其中以500μL样本稀释液4℃过夜洗脱后的DBS检测结果与血浆等效性最佳,71对临床配对样本的检测一致性为98.59%(70/71)。结论 DBS样本可用于HIV-1新发感染检测(限制性抗原亲和力法),且与血浆样本具有良好的等效性。

Objective To study the feasibility and the best test conditions of DBS samples for the detection of recent HIV-1 infection(limiting antigen avidity enzyme immunoassay, Lag-Avidity EIA). Methods Three sets of serial dilution samples of DBS-plasma matched samples were prepared and 71 samples of DBS-plasma matched samples from clinical infected persons were collected. The domestic recent HIV-1 infection detection kit(Lag-Avidity EIA) was used to detect the samples, and analysis was made for the samples equivalence. The DBS samples(6 mm spots) were eluted at 4℃ overnight in 500μL, 600μL, 800μL and 1 000μL diluted solution(in the kit) respectively. Results The results of serial dilution DBS-plasma matched samples showed that the pearson correlation coefficients of the DBS samples, eluted in 500μL, 600μL, 800μL and 1 000μL, and plasma samples were 0.955(R^2=0.912), 0.944(R^2=0.892), 0.948(R^2=0.899), and 0.933(R^2=0.936), and the results of 71 DBS-plasma matched samples from clinical infected persons were 0.968(R^2=0.936), 0.965(R^2=0.932), 0.959(R^2=0.919), and 0.879(R^2=0.773), respectively. The concordance rate between the two types of specimens was 98.59%(70/71).DBS after the elution conditions of 500μL sample diluents at 4℃ overnight had the best equivalence testing results with plasma. Conclusion DBS samples can be used to detect recent HIV-1 infection(Lag-Avidity EIA), and have good equivalence with matched plasma samples.