Short Report: Comparison of the Mosquito Inoculation Technique and Quantitative Real Time Polymerase Chain Reaction to Measure Dengue Virus Concentration
Short Report: Comparison of the Mosquito Inoculation Technique and Quantitative Real Time Polymerase Chain Reaction to Measure Dengue Virus Concentration
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DOI：
10.4269/ajtmh.13-0100
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发表时间：
2013-11-01
影响因子：
3.3
通讯作者：
Gubler, Duane J.
中科院分区：
文献类型：
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作者：
Choy, Milly M.;Ellis, Brett R.;Gubler, Duane J.
准确测量人类和蚊子组织中的传染性登革热病毒对于充分了解病毒与宿主的关系、疾病严重程度、病毒适应性和发病机制至关重要。近年来，通过定量实时聚合酶链式反应测量的RNA拷贝数已被用于测量体外和体内登革热病毒浓度。在这项研究中，我们详细介绍了使用 RNA 基因组当量和蚊子感染剂量 50 (MID50) 测量 Vero 和 C6/36 组织培养物、埃及伊蚊和病毒血症人类血清中病毒生长动力学的重要差异。尽管两种方法之间存在相当好的相关性，但 RNA 拷贝数比传染性病毒滴度高 2 至 5 个对数。这些差异根据病毒株、病毒平台、感染性病毒检测和病毒生长阶段的不同而显着变化。这些结果对于正确解释实验和临床研究的生物学和流行病学数据具有重要意义，并表明在此类研究中将基因组等效物用作传染性病毒的替代物时应谨慎解释。
An accurate measure of infectious dengue virus in human and mosquito tissues is critical to fully understand virus-host relationships, disease severity, viral fitness, and pathogenesis. In recent years, RNA copy number measured by quantitative real time-polymerase chain reaction has been used to measure dengue virus concentration in vitro and in vivo. In this study, we detail important differences in the measurement of viral growth kinetics in Vero and C6/36 tissue cultures, in Aedes aegypti mosquitoes, and in viremic human sera using RNA genomic equivalents and mosquito infectious dose 50 (MID50). Although there was reasonably good correlation between the two methods, RNA copy number was 2 to 5 logs greater than infectious virus titers. These differences varied significantly depending on virus strain, viral platform, infectious virus assay, and viral growth phase. The results have important implications for the correct interpretation of biological and epidemiological data from experimental and clinical studies, and show that genomic equivalents should be interpreted with caution when used as a proxy for infectious virus in such studies.