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评估核酸稳定产品用于在室温下运输和储存干燥全血形式的病毒RNA和抗体的效果。

Evaluation of nucleic acid stabilization products for ambient temperature shipping and storage of viral RNA and antibody in a dried whole blood format.

作者信息

Dauner Allison L, Gilliland Theron C, Mitra Indrani, Pal Subhamoy, Morrison Amy C, Hontz Robert D, Wu Shuenn-Jue L

出版信息

Am J Trop Med Hyg. 2015 Jul;93(1):46-53. doi: 10.4269/ajtmh.15-0110. Epub 2015 May 4.

Abstract

Loss of sample integrity during specimen transport can lead to false-negative diagnostic results. In an effort to improve upon the status quo, we used dengue as a model RNA virus to evaluate the stabilization of RNA and antibodies in three commercially available sample stabilization products: Whatman FTA Micro Cards (GE Healthcare Life Sciences, Pittsburgh, PA), DNAstāble Blood tubes (Biomātrica, San Diego, CA), and ViveST tubes (ViveBio, Alpharetta, GA). Both contrived and clinical dengue-positive specimens were stored on these products at ambient temperature or 37°C for up to 1 month. Antibody and viral RNA levels were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays, respectively, and compared with frozen unloaded controls. We observed reduced RNA and antibody levels between stabilized contrived samples and frozen controls at our earliest time point, and this was particularly pronounced for the FTA cards. However, despite some time and temperature dependent loss, a 94.6-97.3% agreement was observed between stabilized clinical specimens and their frozen controls for all products. Additional considerations such as cost, sample volume, matrix, and ease of use should inform any decision to incorporate sample stabilization products into a diagnostic testing workflow. We conclude that DNAstāble Blood and ViveST tubes are useful alternatives to traditional filter paper for ambient temperature shipment of clinical specimens for downstream molecular and serological testing.

摘要

样本运输过程中样本完整性的丧失可能导致假阴性诊断结果。为了改善现状,我们以登革热作为模型RNA病毒,评估了三种市售样本稳定产品中RNA和抗体的稳定性:Whatman FTA Micro Cards(通用电气医疗生命科学公司,宾夕法尼亚州匹兹堡)、DNAstāble Blood tubes(Biomātrica公司,加利福尼亚州圣地亚哥)和ViveST tubes(ViveBio公司,佐治亚州阿尔法利塔)。将人为制备的和临床登革热阳性样本在这些产品上于室温或37°C下保存长达1个月。分别通过酶联免疫吸附测定(ELISA)和定量逆转录聚合酶链反应(qRT-PCR)测定抗体和病毒RNA水平,并与未加载的冷冻对照进行比较。在最早的时间点,我们观察到稳定的人为制备样本与冷冻对照之间的RNA和抗体水平降低,这在FTA卡上尤为明显。然而,尽管存在一些时间和温度依赖性损失,但所有产品的稳定临床样本与其冷冻对照之间的一致性为94.6 - 97.3%。在决定将样本稳定产品纳入诊断测试工作流程时,应考虑成本、样本体积、基质和易用性等其他因素。我们得出结论,对于临床样本在室温下运输以进行下游分子和血清学检测而言,DNAstāble Blood tubes和ViveST tubes是传统滤纸的有用替代品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6cc1/4497903/53c2a56a8c42/tropmed-93-46-g001.jpg

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