High-throughput multiplexed xMAP Luminex array panel for detection of twenty two medically important mosquito-borne arboviruses based on innovations in synthetic biology.
High-throughput multiplexed xMAP Luminex array panel for detection of twenty two medically important mosquito-borne arboviruses based on innovations in synthetic biology.
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DOI：
10.1016/j.jviromet.2015.01.003
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发表时间：
2015-03
影响因子：
3.1
通讯作者：
Benner SA
中科院分区：
文献类型：
--
作者：
Glushakova LG;Bradley A;Bradley KM;Alto BW;Hoshika S;Hutter D;Sharma N;Yang Z;Kim MJ;Benner SA
蚊媒虫媒病毒作为重要的人类和动物病原体正在全球范围内出现。这使得准确、快速识别它们的分析对于公共卫生、流行病学、生态研究至关重要。在过去的十年中，已经报道了许多针对虫媒病毒核酸的单一和多重测定。目前还没有建立在“单管”环境中常规鉴定多种病毒的方法。随着多重检测的增加，病毒 RNA 的检测因噪声、假阳性和阴性而变得复杂。在这项研究中，开发了一种检测方法，通过结合合成生物学领域出现的两种新型核酸来避免这些问题。第一个是“自我回避分子识别系统”（SAMRS），它可以实现高水平的多重分析。第二个是“人工扩展遗传信息系统”（AEGIS），它能够以嵌套 PCR 格式进行干净的 PCR 扩增。使用转换技术将AEGIS组件放入扩增子中，提高了它们在Luminex珠子上的杂交效率。当 Luminex“液体微阵列”用于下游检测时，该组合支持单管 PCR 扩增测定，可识别黄病毒属、甲病毒属、正布尼亚病毒属的 22 种蚊媒 RNA 病毒。该检测可区分密切相关的病毒，如登革热、西尼罗河病毒、日本脑炎和加州血清学病毒。使用登革热病毒和受感染的蚊子评估了该测定的性能和灵敏度；一只蚊子中可检测到 6-10 个登革热病毒粒子。
Mosquito-borne arboviruses are emerging world-wide as important human and animal pathogens. This makes assays for their accurate and rapid identification essential for public health, epidemiological, ecological studies. Over the past decade, many mono- and multiplexed assays targeting arboviruses nucleic acids have been reported. None has become established for the routine identification of multiple viruses in a “single tube” setting. With increasing multiplexing, the detection of viral RNAs is complicated by noise, false positives and negatives. In this study, an assay was developed that avoids these problems by combining two new kinds of nucleic acids emerging from the field of synthetic biology. The first is a “self-avoiding molecular recognition system” (SAMRS), which enables high levels of multiplexing. The second is an “artificially expanded genetic information system” (AEGIS), which enables clean PCR amplification in nested PCR formats. A conversion technology was used to place AEGIS component into amplicon, improving their efficiency of hybridization on Luminex beads. When Luminex “liquid microarrays” are exploited for downstream detection, this combination supports single-tube PCR amplification assays that can identify 22 mosquito-borne RNA viruses from the genera Flavivirus, Alphavirus, Orthobunyavirus. The assay differentiates between closely-related viruses, as dengue, West Nile, Japanese encephalitis, and the California serological group. The performance and the sensitivity of the assay were evaluated with dengue viruses and infected mosquitoes; as few as 6–10 dengue virions can be detected in a single mosquito.
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DOI：
10.1002/anie.201001977
发表时间：
2010-07-26
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Hoshika S;Chen F;Leal NA;Benner SA
通讯作者：
Benner SA
DOI：
10.4269/ajtmh.2010.09-0676
发表时间：
2010-05-01
影响因子：
3.3
作者：
Hills, Susan L.;Griggs, Anne C.;Fischer, Marc
通讯作者：
Fischer, Marc
影响因子：
3.1
作者：
Houng, HSH;Chen, RCM;Kanesa-thasan, N
通讯作者：
Kanesa-thasan, N
DOI：
10.1098/rspb.2007.1497
发表时间：
2008-02-22
影响因子：
4.7
作者：
Alto, Barry W.;Lounibos, L. Philip;Reiskind, Michael H.
通讯作者：
Reiskind, Michael H.
影响因子：
11.8
作者：
Gubler, Duane J.
通讯作者：
Gubler, Duane J.