Marconi A, Balestrieri M, Comastri G, Pulvirenti F R, Gennari W, Tagliazucchi S, Pecorari M, Borghi V, Marri D, Zazzi M
Department of Molecular Biology, University of Siena, Siena, Italy.
Clin Microbiol Infect. 2009 Jan;15(1):93-7. doi: 10.1111/j.1469-0691.2008.02116.x.
The Abbott Real-Time HIV-1 assay was evaluated for its performance in quantification of human immunodeficiency virus type 1 (HIV-1) RNA in dried blood spot (DBS) samples. In total, 169 blood samples with detectable plasma HIV-1 RNA were used to extract RNA from paired DBS and liquid plasma samples, using the automated Abbott m Sample Preparation System (m2000sp). HIV-1 RNA was then quantitated by the m2000rt RealTime analyser. RNA samples suitable for real-time PCR were obtained from all but one (99.4%) of the DBS samples and HIV-1 RNA was detected in 163/168 (97.0%) samples. The correlation between HIV-1 RNA values measured in paired DBS and plasma samples was very high (r = 0.882), with 78.5% and 99.4% of cases differing by <0.5 and 1.0 log, respectively. Retesting of DBS replicates following 6 months of storage at 2-8 degrees C showed no loss of HIV-1 RNA in a subset of 89 samples. The feasibility of DBS testing coupled with automated sample processing, and the use of a latest-generation FDA-approved real-time PCR-based system, represents an encouraging first step for viral load measurement in reference centres in developing countries where access to antiretroviral therapy is expanding.
对雅培实时HIV-1检测法在定量干血斑(DBS)样本中的1型人类免疫缺陷病毒(HIV-1)RNA方面的性能进行了评估。总共使用169份血浆中可检测到HIV-1 RNA的血样,通过雅培自动化m样本制备系统(m2000sp)从配对的DBS和液体血浆样本中提取RNA。然后用m2000rt实时分析仪对HIV-1 RNA进行定量。除一份样本外,从所有(99.4%)DBS样本中均获得了适合实时PCR的RNA样本,并且在168份样本中的163份(97.0%)中检测到了HIV-1 RNA。配对的DBS和血浆样本中测得的HIV-1 RNA值之间的相关性非常高(r = 0.882),分别有78.5%和99.4%的病例差异<0.5和1.0 log。在2-8摄氏度下储存6个月后的DBS重复样本重新检测显示,在89份样本的子集中HIV-1 RNA没有损失。DBS检测结合自动化样本处理以及使用最新一代FDA批准的基于实时PCR的系统的可行性,对于抗逆转录病毒疗法可及性正在扩大的发展中国家的参考中心进行病毒载量测量来说,是鼓舞人心的第一步。