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使用自动雅培实时病毒载量测定法,比较血浆和干血斑样本中 HIV-1 RNA 的测量值。

Comparison of HIV-1 RNA measurements obtained by using plasma and dried blood spots in the automated abbott real-time viral load assay.

机构信息

Department of Infectious Diseases, Hospital Carlos III, Madrid, Spain.

出版信息

J Clin Microbiol. 2012 Mar;50(3):569-72. doi: 10.1128/JCM.00418-11. Epub 2011 Dec 14.

Abstract

Dried blood spots (DBS) may be a promising alternative specimen type to plasma for measuring the viral load (VL) in HIV-infected individuals in resource-limited settings. However, characterization of assay performance using DBS is incomplete. In this prospective study, the VL was measured in parallel using plasma and DBS specimens collected at the same time from 157 HIV-1-infected individuals. DBS were prepared by dispensing 50 μl of blood onto filter paper cards and were stored desiccated at -20°C. Nucleic acid extraction from plasma and DBS was performed automatically using the Abbott m2000sp instrument, and the VL was measured using the RealTime HIV-1 VL assay, which has a lower limit of detection of 40 HIV RNA copies/ml. The correlation between plasma and DBS results was good (R = 0.91; P < 0.001). The mean difference in the VL (DBS minus plasma) was 0.35 log copies (standard deviation [SD], 0.47 log copies). A total of 40 (26%) paired specimens had a difference of >0.5 log copy, and in 12 (7.8%) it was >1 log copy. the VL from DBS was measurable in 95.7% of specimens with a plasma VL of >2.74 log copies (550 HIV RNA copies/ml). In summary, the VL can reliably be measured using DBS with the Abbott RealTime HIV-1 assay. The estimated lower limit of detection of this automated methodology on DBS is 550 copies/ml, a threshold that may be acceptable for periodic VL monitoring in patients on antiretroviral therapy in resource-limited settings, where early detection of virologic treatment failure is often problematic.

摘要

干血斑(DBS)可能是一种有前途的替代血浆样本类型,可用于在资源有限的环境中测量感染 HIV 的个体的病毒载量(VL)。然而,使用 DBS 对检测性能进行特征描述并不完整。在这项前瞻性研究中,同时使用从 157 名 HIV-1 感染个体采集的血浆和 DBS 标本平行测量 VL。DBS 通过将 50μl 的血液分配到滤纸上的卡片上制备,并在 -20°C 下干燥储存。使用 Abbott m2000sp 仪器自动从血浆和 DBS 中提取核酸,并使用 RealTime HIV-1 VL 测定法测量 VL,该方法的检测下限为 40 HIV RNA 拷贝/ml。血浆和 DBS 结果之间的相关性良好(R = 0.91;P <0.001)。VL(DBS 减去血浆)的平均差值为 0.35 log 拷贝(标准偏差[SD]为 0.47 log 拷贝)。共有 40 对(26%)配对标本的差值> 0.5 log 拷贝,其中 12 对(7.8%)差值> 1 log 拷贝。在血浆 VL > 2.74 log 拷贝(550 HIV RNA 拷贝/ml)的 95.7%的标本中,DBS 中的 VL 可测量。总之,使用 Abbott RealTime HIV-1 测定法可以可靠地测量 DBS 中的 VL。该自动方法在 DBS 上的估计检测下限为 550 拷贝/ml,对于资源有限环境中接受抗逆转录病毒治疗的患者进行定期 VL 监测,这可能是可接受的,因为早期检测病毒学治疗失败通常是个问题。

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