Titchmarsh Logan, Zeh Clement, Verpoort Thierry, Allain Jean-Pierre, Lee Helen
Diagnostic Development Unit, Department of Haematology, University of Cambridge, Cambridge, United Kingdom.
U.S. Centers for Disease Control and Prevention, HIV-Research Branch, Kisumu, Kenya, and Division of HIV/AIDS Prevention, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
J Clin Microbiol. 2015 Apr;53(4):1080-6. doi: 10.1128/JCM.02853-14. Epub 2014 Nov 26.
In order to limit the interference of HIV-1 cellular nucleic acids in estimating viral load (VL), the feasibility of leukodepletion of a small whole-blood (WB) volume to eliminate only leukocyte cell content was investigated, using a selection of filters. The efficacy of leukocyte filtration was evaluated by counting, CD45 quantitative PCR, and HIV-1 DNA quantification. Plasma HIV-1 was tested by real-time reverse transcription (RT)-PCR. A specific, miniaturized filter was developed and tested for leukocyte and plasma virus retention, WB sample dilution, and filtration parameters in HIV-1-spiked WB samples. This device proved effective to retain >99.9% of white blood cells in 100 μl of WB without affecting plasma VL. The Samba sample preparation chemistry was adapted to use a leukodepleted WB sample for VL monitoring using the point-of-care Samba-1 semiautomated system. The clinical performance of the assay was evaluated by testing 207 consecutive venous EDTA WB samples from HIV-1-infected patients attending a CD4 testing clinic. Most patients were on antiretroviral treatment (ART), but their VL status was unknown. Compared to the Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test, the new Samba assay had a concordance of 96.5%. The use of the Samba system with a VL test for WB might contribute to HIV-1 ART management and reduce loss-to-follow-up rates in resource-limited settings.
为了限制HIV-1细胞核酸对病毒载量（VL）估计的干扰，研究了使用一系列滤器对少量全血（WB）进行白细胞去除以仅消除白细胞细胞成分的可行性。通过计数、CD45定量PCR和HIV-1 DNA定量来评估白细胞过滤的效果。血浆HIV-1通过实时逆转录（RT）-PCR进行检测。开发并测试了一种特定的小型化滤器，用于检测HIV-1加标的WB样本中的白细胞和血浆病毒保留情况、WB样本稀释情况以及过滤参数。该装置被证明可有效保留100 μl WB中>99.9%的白细胞，且不影响血浆VL。对Samba样本制备化学方法进行了调整，以便使用白细胞去除后的WB样本通过即时护理Samba-1半自动系统进行VL监测。通过检测来自一家CD4检测诊所的207例连续HIV-1感染患者的静脉EDTA WB样本，评估了该检测方法的临床性能。大多数患者正在接受抗逆转录病毒治疗（ART），但他们的VL状态未知。与罗氏Cobas AmpliPrep/Cobas TaqMan HIV-1检测相比，新的Samba检测方法的一致性为96.5%。将Samba系统与WB的VL检测方法结合使用可能有助于HIV-1的ART管理，并降低资源有限环境中的失访率。
