Lloyd R M, Burns D A, Huong J T, Mathis R L, Winters M A, Tanner M, De La Rosa A, Yen-Lieberman B, Armstrong W, Taege A, McClernon D R, Wetshtein J L, Friedrich Brian M, Ferguson Monique R, O'Brien William, Feorino P M, Holodniy M
Research Think Tank, Inc., Buford, Georgia, USA.
J Clin Microbiol. 2009 May;47(5):1491-6. doi: 10.1128/JCM.02354-08. Epub 2009 Mar 25.
A novel method for the collection and transportation of dried-blood-plasma samples, SampleTanker (ST), was developed and compared to standard shipping protocols for frozen-plasma specimens containing human immunodeficiency virus type 1 (HIV-1) and/or hepatitis C virus (HCV). Matched frozen and dried 1-ml EDTA-containing plasma samples were collected and analyzed by several molecular-based virologic assays. After addition of 1.175 ml of reconstitution buffer, 1.035 ml of dried plasma was recovered. Mean intra-assay variances were 0.05, 0.05, and 0.06 log(10) copies/ml for the Versant, Amplicor, and NucliSens QT HIV-1 load assays, respectively (P, not significant). However, mean HIV-1 viral load was consistently reduced in dried samples by 0.32 to 0.51 log(10) copies/ml, depending on assay type (P < 0.05). Infectious HIV-1 was not recovered from dried ST plasma. There was no significant difference in HIV-1 viral load results obtained using ST after 8 weeks of storage at ambient temperature. Compared to frozen plasma, HIV-1 genotypic results were >99% concordant at the nucleotide and amino acid levels, as well as for resistance-associated mutations. We further demonstrated successful detection of multiple analytes, including HIV-1 viral load, HIV-1 antiretroviral resistance genotype, and HCV genotype, from a single ST unit. Dried plasma collected with ST yielded comparable results to frozen samples for multiple-analyte clinical testing. As such, ST could be a useful alternative for virologic tests and clinical trials worldwide by significantly diminishing transportation cost and the sample volume restrictions associated with dried-blood-spot technology.
开发了一种用于收集和运输干血血浆样本的新方法——样本运输器(SampleTanker,ST),并将其与用于含有1型人类免疫缺陷病毒(HIV-1)和/或丙型肝炎病毒(HCV)的冷冻血浆标本的标准运输方案进行比较。收集了匹配的1ml含乙二胺四乙酸(EDTA)的冷冻和干血浆样本,并通过几种基于分子的病毒学检测方法进行分析。加入1.175ml重构缓冲液后,回收了1.035ml干血浆。对于Versant、Amplicor和NucliSens QT HIV-1载量检测,平均批内方差分别为0.05、0.05和0.06 log(10)拷贝/ml(P,无显著性差异)。然而,根据检测类型,干样本中的HIV-1病毒载量平均持续降低0.32至0.51 log(10)拷贝/ml(P<0.05)。未从干的ST血浆中回收有传染性的HIV-1。在室温下储存8周后,使用ST获得的HIV-1病毒载量结果没有显著差异。与冷冻血浆相比,HIV-1基因分型结果在核苷酸和氨基酸水平以及耐药相关突变方面的一致性>99%。我们进一步证明了从单个ST单元成功检测多种分析物,包括HIV-1病毒载量、HIV-1抗逆转录病毒耐药基因型和HCV基因型。用ST收集的干血浆在多分析物临床检测中产生的结果与冷冻样本相当。因此,通过显著降低运输成本和与干血斑技术相关的样本量限制,ST可能成为全球病毒学检测和临床试验的有用替代方法。