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与干血斑(DBS)标本相比,全血中 HIV-1 核酸的稳定性提高,并且可以更有效地检测到其他类型的替代标本中的 HIV-1。

The stability of HIV-1 nucleic acid in whole blood and improved detection of HIV-1 in alternative specimen types when compared to Dried Blood Spot (DBS) specimens.

机构信息

Rush Medical College, Department of Microbial Pathogens and Immunity, Chicago, IL, USA.

New England Research Institute, Boston, MA, USA.

出版信息

J Virol Methods. 2018 Nov;261:91-97. doi: 10.1016/j.jviromet.2018.08.009. Epub 2018 Aug 17.

Abstract

BACKGROUND

Commercially-available kits for HIV-1 detection include instructions for detecting HIV-1 in plasma and DBS, but don't support other specimen types.

OBJECTIVES

Show quantitative stability of HIV-1 total nucleic acid (TNA) in blood and improved HIV-1 detection in alternative specimen types.

STUDY DESIGN

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.

RESULTS

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).

CONCLUSIONS

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.

摘要

背景

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。

目的

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。

研究设计

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。

结果

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)。

结论

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。

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