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塞内加尔通过实时NucliSENS EasyQ HIV-1检测法对干血斑中的HIV-1 RNA进行定量分析。

Quantitation of HIV-1 RNA in dried blood spots by the real-time NucliSENS EasyQ HIV-1 assay in Senegal.

作者信息

Kane Coumba Toure, Ndiaye Halimatou Diop, Diallo Sada, Ndiaye Ibrahima, Wade Abdoulaye Sidibé, Diaw Papa Alassane, Gaye-Diallo Aïssatou, Mboup Souleymane

机构信息

Bacteriology-Virology Laboratoire de CHU A. Le Dantec, Dakar, Senegal.

出版信息

J Virol Methods. 2008 Mar;148(1-2):291-5. doi: 10.1016/j.jviromet.2007.11.011. Epub 2008 Feb 1.

Abstract

Measurement of viral load in plasma remains the best marker for the follow-up of antiretroviral therapy. However, its use is limited in developing countries due to the lack of adequate facilities and equipment, and cryopreservation of plasma during storage and transportation. Practical and reliable methods adapted to field conditions for the collection, transportation and accurate measurement of HIV-1 viral load are needed for the optimum use of antiretroviral therapy in resource-limited countries. This study evaluated the use of dried blood spots (DBS) for the real-time quantitation of HIV-1 RNA levels with the NucliSENS EasyQ((R)) HIV-1 assay (bioMérieux, Lyon, France) under field conditions in Senegal (Africa). Dried blood spots and plasma from 41 patients living in suburban Dakar were used for determination of HIV-1 RNA concentrations and stability at 37 degrees C. Analysis was performed at the Dakar University Hospital laboratory. Extraction was done with the bioMérieux NucliSENS((R)) miniMAGtrade mark, and real-time detection was done with the bioMérieux NucliSENS((R)) EasyQ system. HIV-1 RNA concentrations in plasma were compared with concentrations in dried blood spots after 8 and 15 days at 37 degrees C. The study showed a strong concordance in RNA levels between plasma and dried blood spots, which appear to be very stable over time with no apparent degradation observed after 2 weeks at 37 degrees C (mean difference 0.065logIU/ml). These results suggest that the use of dried blood spots in combination with the NucliSENS EasyQ HIV-1 assay is well adapted for HIV-1 RNA level monitoring in centralized laboratories in developing countries.

摘要

血浆中病毒载量的测定仍然是抗逆转录病毒治疗随访的最佳指标。然而,由于缺乏足够的设施和设备,以及血浆在储存和运输过程中的冷冻保存,其在发展中国家的应用受到限制。在资源有限的国家,为了最佳地使用抗逆转录病毒治疗,需要适合现场条件的实用且可靠的方法来收集、运输和准确测量HIV-1病毒载量。本研究在非洲塞内加尔的现场条件下,评估了使用干血斑(DBS)通过NucliSENS EasyQ((R)) HIV-1检测法(法国里昂生物梅里埃公司)对HIV-1 RNA水平进行实时定量。来自达喀尔郊区41名患者的干血斑和血浆用于测定HIV-1 RNA浓度以及在37摄氏度下的稳定性。分析在达喀尔大学医院实验室进行。提取使用生物梅里埃NucliSENS((R)) miniMAG商标产品,实时检测使用生物梅里埃NucliSENS((R)) EasyQ系统。将血浆中HIV-1 RNA浓度与在37摄氏度下保存8天和15天后干血斑中的浓度进行比较。研究表明,血浆和干血斑中的RNA水平高度一致,随着时间推移似乎非常稳定,在37摄氏度下放置2周后未观察到明显降解(平均差异0.065logIU/ml)。这些结果表明,干血斑与NucliSENS EasyQ HIV-1检测法联合使用非常适合发展中国家集中实验室监测HIV-1 RNA水平。

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