Department of Therapeutic Research and Medicines Evaluation, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
J Clin Virol. 2010 Jan;47(1):4-7. doi: 10.1016/j.jcv.2009.11.006. Epub 2009 Dec 4.
The use of simplified methods for viral load determination could greatly increase access to treatment monitoring of HIV patients in resource-limited countries.
The aim of the present study was to optimize and evaluate the performance of the Roche COBAS Taqman assay in HIV-RNA quantification from dried blood spots (DBS) and dried plasma spots (DPS).
EDTA blood samples from 108 HIV-infected women were used to prepare 129 DBS and 76 DPS on Whatman 903 card. DBS and DPS were stored at -20 degrees C. HIV-1 RNA was extracted from DBS/DPS using the MiniMAG system (bioMerieux). Amplification and detection were performed using the Roche COBAS TaqMan assay. Plasma viral load results were used as standard.
There was a high correlation between measures of viral load in plasma and in DBS/DPS (r=0.96 and 0.85 respectively, P<0.001). Overall, viral load values in DBS and DPS tended to be lower than in plasma with mean (SD) differences of 0.32 log(0.22) for DBS and of 0.35 (0.33) for DPS. Detection rates were 96.4% for DBS and 96.1% for DPS in samples with corresponding plasma values >3.0 log copies/ml. Samples with HIV-RNA below 50 copies/ml were correctly identified in 18/19 DBS and in 7/7 DPS.
Both DBS and DPS provided results highly correlated to the plasma values. High detection rate was obtained with both DBS and DPS when HIV-RNA was >3.0 log copies/ml. Our results support the use of DBS/DPS to detect virologic failure in resource-limited settings.
在资源有限的国家中,使用简化方法进行病毒载量测定可以极大地增加对 HIV 患者治疗监测的可及性。
本研究旨在优化和评估罗氏 COBAS Taqman 检测法在从干血斑(DBS)和干血浆斑(DPS)中定量 HIV-RNA 方面的性能。
使用 EDTA 抗凝血液样本,从 108 名 HIV 感染妇女中制备了 129 个 DBS 和 76 个 DPS,均在 Whatman 903 卡上。DBS 和 DPS 储存在-20°C。使用 MiniMAG 系统(bioMerieux)从 DBS/DPS 中提取 HIV-1 RNA。使用罗氏 COBAS TaqMan 检测法进行扩增和检测。将血浆病毒载量结果作为标准。
血浆和 DBS/DPS 中病毒载量的测量值高度相关(r=0.96 和 0.85,均 P<0.001)。总体而言,DBS 和 DPS 中的病毒载量值倾向于低于血浆值,DBS 的平均(SD)差值为 0.32 log(0.22),DPS 的差值为 0.35(0.33)。对于对应血浆值>3.0 log 拷贝/ml 的样本,DBS 的检测率为 96.4%,DPS 的检测率为 96.1%。在 HIV-RNA<50 拷贝/ml 的样本中,19 个 DBS 和 7 个 DPS 中的样本均得到正确识别。
DBS 和 DPS 均提供与血浆值高度相关的结果。当 HIV-RNA>3.0 log 拷贝/ml 时,DBS 和 DPS 均获得了高检测率。我们的研究结果支持在资源有限的环境中使用 DBS/DPS 来检测病毒学失败。