VA Palo Alto Health Care System, Palo Alto, CA, USA.
Influenza Other Respir Viruses. 2011 Nov;5(6):413-7. doi: 10.1111/j.1750-2659.2011.00253.x. Epub 2011 Apr 18.
Most clinical samples collected for diagnostic influenza testing and monitoring require refrigerated or frozen storage or shipment, which imparts logistic and cost burdens. The ability to store and ship dried clinical specimens under ambient conditions for influenza testing would significantly reduce costs and protect samples from improper storage or equipment failure, especially in remote or resource-limited areas.
To evaluate the collection and storage of dried clinical samples on a transport matrix (ViveST™, ST) for influenza RNA testing by real-time reverse-transcription PCR (RT-PCR).
Viral transport medium from swab or sputum samples was applied to ST, dried, and stored under ambient conditions from 2 days to 6 months. Additional aliquots of samples were frozen. Testing of frozen and ST-stored samples was performed using the WHO/CDC real-time influenza A (H1N1) RT-PCR protocol and compared to the Luminex xTAG RVP assay.
ST-stored samples yielded slightly higher threshold cycle values (median 2·54 cycles) compared to frozen samples tested in parallel. This difference was consistent regardless of viral input. There was no significant difference in signal recovery between samples stored for 1 week versus samples stored for 3 weeks, or from three samples stored for 6 months. Qualitatively, clinical specimens stored on ST were 100% concordant (36/36) with frozen samples for detecting the presence of influenza A RNA.
ST-processed dried specimens produced similar rates of seasonal or novel 2009 HIN1 influenza RNA detection compared to conventional sample processing and thus presents a viable alternative to refrigerated or frozen samples.
大多数用于诊断流感检测和监测的临床样本需要冷藏或冷冻保存或运输,这带来了物流和成本负担。能够在环境条件下储存和运输干燥的临床标本进行流感检测,将显著降低成本,并防止样本因储存不当或设备故障而受损,特别是在偏远或资源有限的地区。
评估在运输基质(ViveST™,ST)上采集和储存干燥临床样本用于实时逆转录聚合酶链反应(RT-PCR)检测流感 RNA 的效果。
将拭子或痰样本中的病毒运输介质涂抹在 ST 上,干燥后在环境条件下储存 2 天至 6 个月。对样本进行等分冷冻。使用世界卫生组织/疾病控制与预防中心的实时流感 A(H1N1)RT-PCR 方案对冷冻和 ST 储存的样本进行检测,并与 Luminex xTAG RVP 检测进行比较。
与平行检测的冷冻样本相比,ST 储存的样本产生的阈值循环值(中位数 2.54 个循环)略高。无论病毒输入量如何,这种差异都是一致的。在 1 周和 3 周储存的样本之间,或在 6 个月内储存的 3 个样本之间,信号恢复没有显著差异。定性而言,储存于 ST 的临床标本与冷冻样本在检测流感 A RNA 存在方面完全一致(36/36)。
与传统样本处理相比,ST 处理的干燥标本在检测季节性或新型 2009 年 H1N1 流感 RNA 方面的成功率相似,因此是冷藏或冷冻样本的可行替代方法。