Rush Medical College, Department of Microbial Pathogens and Immunity, Chicago, IL, USA.
New England Research Institute, Boston, MA, USA.
J Virol Methods. 2018 Nov;261:91-97. doi: 10.1016/j.jviromet.2018.08.009. Epub 2018 Aug 17.
Commercially-available kits for HIV-1 detection include instructions for detecting HIV-1 in plasma and DBS, but don't support other specimen types.
Show quantitative stability of HIV-1 total nucleic acid (TNA) in blood and improved HIV-1 detection in alternative specimen types.
Whole blood and DBS specimens, tested as part of an external quality assurance program for qualitative HIV-1 detection, were used to evaluated error rates (false negative [FN], false positive [FP] and indeterminant [IND] results) across assays (internally developed [IH], Roche Amplicor [RA], and Roche TaqMan Qual [TQ]) and specimen types (frozen whole blood [BLD], DBS and cell pellets [PEL]). A modified Roche TaqMan HIV-1 assay was used to quantify HIV-1 TNA.
Significantly higher error rates were noted in DBS across all of the assays (4% vs. 0% for DBS and PEL, IH, p = 0.005; 4% vs. 0.1% for DBS and PEL, RA, p < 0.001; 10% vs. 1% for DBS and PEL or BLD, TQ, p < 0.001). HIV TNA concentration is stable in BLD (day 1 vs. day 10, p = 0.39) and higher than DBS (p < 0.001).
Transporting refrigerated whole blood for centralized processing into alternative specimen types will improve the sensitivitiy of HIV-1 detection in samples with low virus loads.
市售的 HIV-1 检测试剂盒包括用于检测血浆和 DBS 中 HIV-1 的说明,但不支持其他样本类型。
展示血液中 HIV-1 总核酸(TNA)的定量稳定性,并提高替代样本类型中 HIV-1 的检测灵敏度。
全血和 DBS 标本作为定性 HIV-1 检测外部质量保证计划的一部分进行检测,用于评估不同检测方法(内部开发[IH]、罗氏 Amplicor [RA]和罗氏 TaqMan Qual [TQ])和标本类型(冷冻全血[BLD]、DBS 和细胞沉淀[PEL])之间的错误率(假阴性[FN]、假阳性[FP]和不确定[IND]结果)。使用改良的罗氏 TaqMan HIV-1 检测法来定量 HIV-1 TNA。
在所有检测方法中,DBS 的错误率显著更高(DBS 和 PEL 与 IH 相比,4%比 0%,p=0.005;DBS 和 PEL 与 RA 相比,4%比 0.1%,p<0.001;DBS 和 PEL 或 BLD 与 TQ 相比,10%比 1%,p<0.001)。BLD 中的 HIV TNA 浓度稳定(第 1 天与第 10 天相比,p=0.39),且高于 DBS(p<0.001)。
将冷藏的全血运输到集中处理中心,再转化为替代样本类型,将提高低病毒载量样本中 HIV-1 检测的灵敏度。