Detection of chikungunya viral RNA in mosquito bodies on cationic (Q) paper based on innovations in synthetic biology.
Detection of chikungunya viral RNA in mosquito bodies on cationic (Q) paper based on innovations in synthetic biology.
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DOI：
10.1016/j.jviromet.2017.04.013
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发表时间：
2017-08
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;Alto BW;Kim MS;Bradley A;Yaren O;Benner SA
基孔肯雅病毒（CHIKV）代表了对公共卫生的日益增长的全球关注，其需要廉价且方便的方法来收集作为潜在载体的蚊子，使得它们可以被保存、储存和运输以供以后和/或远程分析。本文报道的是一种纤维素基纸，用季铵基团衍生（“Q纸”），满足这些需求。在一系列测试中，受感染的蚊子尸体被直接压在Q纸上。然后将氨水添加到蚊子身体上以释放病毒RNA，其通过静电相互作用吸附在阳离子表面上。然后将样品储存（冷冻）或运输。为了进行分析，将CHIKV核酸从Q-纸上洗脱，并在先前开发的工作流程中进行PCR扩增，该工作流程还利用了两种核酸创新（“人工扩增遗传信息系统”，AEGIS和“自回避分子识别系统”，SAMRS）。然后通过Luminex杂交测定分析扩增子。该程序在每个感染的蚊子样品中检测到CHIKV RNA（如果存在），但在未感染的对应物或ddH 2 O样品洗涤物中未检测到，在样品收集后一周或十个月进行测试。
Chikungunya virus (CHIKV) represents a growing and global concern for public health that needs inexpensive and convenient methods to collect mosquitoes as potential carriers so that they can be preserved, stored and transported for later and/or remote analysis. Reported here is a cellulose-based paper, derivatized with quaternary ammonium groups (“Q-paper”) that meets these needs. In a series of tests, infected mosquito bodies were squashed directly on to Q-paper. Aqueous ammonia was then added on the mosquito bodies to release viral RNA that adsorbed on the cationic surface via electrostatic interactions. The samples were then stored (frozen) or transported. For analysis, the CHIKV nucleic acids were eluted from the Q-paper and PCR amplified in a workflow, previously developed, that also exploited two nucleic acid innovations, (“artificially expanded genetic information systems”, AEGIS, and “self-avoiding molecular recognition systems”, SAMRS). The amplicons were then analyzed by a Luminex hybridization assay. This procedure detected CHIKV RNA, if present, in each infected mosquito sample, but not in non-infected counterparts or ddH2O samples washes, with testing one week or ten months after sample collection.
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影响因子：
3.1
作者：
Coffey LL;Forrester N;Tsetsarkin K;Vasilakis N;Weaver SC
通讯作者：
Weaver SC
影响因子：
2.1
作者：
Richards, Stephanie L.;Anderson, Sheri L.;Tabachnick, Walter J.
通讯作者：
Tabachnick, Walter J.
影响因子：
2.1
作者：
Benedict, Mark Q.;Levine, Rebecca S.;Lounibos, L. Philip
通讯作者：
Lounibos, L. Philip
DOI：
10.5501/wjv.v1.i1.11
发表时间：
2012-02-12
期刊：
World journal of virology
影响因子：
--
作者：
Devaux, Christian A
通讯作者：
Devaux, Christian A
DOI：
10.1016/j.cccn.2005.06.023
发表时间：
2006-01
期刊：
Clinica chimica acta; international journal of clinical chemistry
影响因子：
--
作者：
Dunbar SA
通讯作者：
Dunbar SA