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42°C短期储存下血浆样本中乙肝病毒DNA的稳定性

Hepatitis B virus DNA stability in plasma samples under short-term storage at 42°C.

作者信息

Almeida R W de, Espírito-Santo M P, Sousa P S F, Almeida A J de, Lampe E, Lewis-Ximenez L L

机构信息

Laboratório de Hepatites Virais, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, RJ, Brasil.

出版信息

Braz J Med Biol Res. 2015 Jun;48(6):553-6. doi: 10.1590/1414-431X20144040. Epub 2015 Mar 13.

Abstract

We evaluated the stability of hepatitis B virus (HBV) DNA in plasma samples stored at 42°C for external quality assessment (EQA) panels of viral load. To assess the stability of plasma samples containing different concentrations of HBV DNA, serial dilutions of HBV-infected samples with a viral load of 6.40 log(10) IU/mL were made to yield viral loads of 5, 4, and 3 log(10) IU/mL. These were incubated at 42°C for up to 7 days and then frozen at -70°C. Viral load testing for HBV DNA was performed for all samples using COBAS® AmpliPrep/COBAS® TaqMan® HBV Test (v.2.0, Roche, Switzerland). Results were compared with fresh frozen plasma samples as a benchmark to establish acceptable measurements on the days following sample collection. Although the results of this study demonstrated a decrease in HBV DNA viral load ranging from 0.005 to 0.30 log(10) IU/mL after storage at 42°C for up to 7 days, these values did not exceed 0.5 log(10), which is the estimated intra-assay variation for molecular tests. Thus, the insignificant decrease in viral load suggests that shipment of HBV in plasma samples at temperatures of up to 42°C is permissible if they are frozen within 7 days.

摘要

我们评估了在42°C储存的血浆样本中乙型肝炎病毒(HBV)DNA的稳定性,用于病毒载量的外部质量评估(EQA)小组。为了评估含有不同浓度HBV DNA的血浆样本的稳定性,对病毒载量为6.40 log(10) IU/mL的HBV感染样本进行系列稀释,以产生病毒载量为5、4和3 log(10) IU/mL的样本。将这些样本在42°C孵育长达7天,然后在-70°C冷冻。使用COBAS® AmpliPrep/COBAS® TaqMan® HBV检测(v.2.0,罗氏公司,瑞士)对所有样本进行HBV DNA病毒载量检测。将结果与新鲜冷冻血浆样本作为基准进行比较,以确定样本采集后几天内可接受的测量值。尽管本研究结果表明,在42°C储存长达7天后,HBV DNA病毒载量下降了0.005至0.30 log(10) IU/mL,但这些值未超过0.5 log(10),这是分子检测的估计批内变异。因此,病毒载量的微小下降表明,如果在7天内冷冻,血浆样本中的HBV在高达42°C的温度下运输是允许的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/99da/4470315/dfe4b004cfd4/1414-431X-bjmbr-48-06-00553-gf01.jpg

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