International Laboratory Branch, Division of Global HIV/AIDS, Center for Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, USA.
J Virol Methods. 2013 Mar;188(1-2):1-5. doi: 10.1016/j.jviromet.2012.11.036. Epub 2012 Dec 7.
Participation in external quality assessment programs is critical to ensure quality clinical laboratory testing. Commercially available proficiency test panels for HIV-1 virus load testing that are used commonly in external quality assessment programs remain a financial obstacle to resource-limited countries. Maintaining cold-chain transportation largely contributes to the cost of traditional liquid proficiency test panels. Therefore, we developed and evaluated a proficiency test panel using dried tube specimens that can be shipped and stored at ambient temperature. This dried tube specimens panel consisted of 20 μl aliquots of a HIV-1 stock that were added to 2 ml tubes and left uncapped for drying, as a preservation method. The stability of dried tube specimens at concentrations ranging from 10² to 10⁶·⁵ RNA copies/ml was tested at different temperatures over time, showing no viral load reduction at 37 °C and a decrease in viral load smaller than 0.5 Log₁₀ at 45 °C for up to eight weeks when compared to initial results. Eight cycles of freezing-thawing had no effect on the stability of the dried tube specimens. Comparable viral load results were observed when dried tube specimen panels were tested on Roche CAPTAQ, Abbott m2000, and Biomerieux easyMAG viral load systems. Preliminary test results of dried proficiency test panels shipped to four African countries at ambient temperature demonstrated a low inter assay variation (SD range: 0.29-0.41 Log₁₀ RNA copies/ml). These results indicated that HIV-1 proficiency test panels generated by this methodology might be an acceptable alternative for laboratories in resource-limited countries to participate in external quality assessment programs.
参与外部质量评估计划对于确保临床实验室检测的质量至关重要。目前，在外部质量评估计划中常用的商业化 HIV-1 病毒载量检测能力验证试剂盒仍然是资源有限国家的一个经济障碍。保持冷链运输在很大程度上增加了传统液体能力验证试剂盒的成本。因此，我们开发并评估了一种使用干燥管标本的能力验证试剂盒，该试剂盒可以在室温下运输和储存。该干燥管标本试剂盒由 20 μl HIV-1 储存液组成，添加到 2 ml 管中，不加盖干燥，作为一种保存方法。在不同温度下，我们测试了浓度范围为 10²至 10⁶·5 RNA 拷贝/ml 的干燥管标本在不同时间的稳定性，结果显示在 37°C 下没有病毒载量减少，在 45°C 下病毒载量下降小于 0.5 Log₁₀，与初始结果相比，最长可达 8 周。八次冻融循环对干燥管标本的稳定性没有影响。当在罗氏 CAPTAQ、雅培 m2000 和生物梅里埃 easyMAG 病毒载量系统上测试干燥管标本试剂盒时，观察到可比的病毒载量结果。当以室温运输到四个非洲国家的干燥能力验证试剂盒的初步测试结果表明，室内检测变异较小（SD 范围：0.29-0.41 Log₁₀ RNA 拷贝/ml）。这些结果表明，通过这种方法生成的 HIV-1 能力验证试剂盒可能是资源有限国家的实验室参与外部质量评估计划的一种可接受的替代方案。
