用于核酸检测的基孔肯雅病毒RNA参考试剂特性的协作研究。

Collaborative study for the characterization of a chikungunya virus RNA reference reagent for use in nucleic acid testing.

作者信息

Añez G, Jiang Z, Heisey D A R, Kerby S, Rios M

机构信息

Office of Blood Research and Review, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Bethesda, MD, USA.

Office of Biostatistics and Epidemiology, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Bethesda, MD, USA.

出版信息

Vox Sang. 2015 Nov;109(4):312-8. doi: 10.1111/vox.12297. Epub 2015 May 26.

DOI:10.1111/vox.12297

PMID:26014282

Abstract

BACKGROUND AND OBJECTIVES

Infections with the mosquito-borne chikungunya virus (CHIKV) can cause febrile illness or be asymptomatic. Laboratory diagnosis of CHIKV is often made with laboratory-developed nucleic acid amplification technology (NAT) assays because there are no U.S. Food and Drug Administration (FDA)-approved diagnostic or blood screening assays. We aimed to produce a well-characterized CHIKV RNA reference reagent (CHIKV-RR) for use in NAT assays.

MATERIALS AND METHODS

A CHIKV RNA-RR consisting of cell culture-grown, heat-inactivated CHIKV diluted in human plasma was assessed by 8 laboratories in a collaborative study. The participants were asked to test the CHIKV-RR using their NAT assay(s) by qualitative testing (determination of RNA end-point by testing log and half-log dilutions followed by calculation of estimated NAT-detectable units/ml, after adjustment for the sample volume used for testing), and by quantitative testing, when available.

RESULTS

Results from the testing showed that the CHIKV-RR had an estimated overall mean of 7.56 log10 detectable units/ml, ranging from 6.2 log10 to 8.6 log10.

CONCLUSIONS

The Center for Biologics for Evaluation and Research/FDA CHIKV RNA-RR for NAT was established with a concentration of 7.56 log10 detectable units/ml.

摘要

背景与目的

蚊媒传播的基孔肯雅病毒（CHIKV）感染可导致发热性疾病或无症状感染。由于美国食品药品监督管理局（FDA）未批准用于诊断或血液筛查的检测方法，CHIKV的实验室诊断通常采用实验室自行开发的核酸扩增技术（NAT）检测方法。我们旨在生产一种特性良好的CHIKV RNA参考试剂（CHIKV-RR）用于NAT检测。

材料与方法

一项合作研究中，8个实验室对一种由在人血浆中稀释的细胞培养生长、热灭活的CHIKV组成的CHIKV RNA-RR进行了评估。要求参与者通过定性检测（通过检测对数和半对数稀释液确定RNA终点，然后在调整用于检测的样品体积后计算估计的NAT可检测单位/毫升）以及在可行时通过定量检测，使用他们的NAT检测方法检测CHIKV-RR。