ABC Assay: Method Development and Application to Quantify the Role of Three DWV Master Variants in Overwinter Colony Losses of European Honey Bees.
ABC Assay: Method Development and Application to Quantify the Role of Three DWV Master Variants in Overwinter Colony Losses of European Honey Bees.
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DOI：
10.3390/v9110314
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发表时间：
2017-10-27
期刊：
影响因子：
--
通讯作者：
Schroeder DC
中科院分区：
文献类型：
--
作者：
Kevill JL;Highfield A;Mordecai GJ;Martin SJ;Schroeder DC
蜜蜂畸形翅病毒（Deformed wing virus，DWV）是世界上最流行的蜜蜂病毒病原之一。DWV是许多RNA病毒的典型，是一种准种，由大量不同的变体组成，目前由三种主要变体组成：A型、B型和C型。关于每个变体或变体组合对个体宿主生物学的影响知之甚少。因此，我们开发了一组新的主变体特异性DWV引物和一组标准品，用于定量每种主变体。竞争性逆转录酶聚合酶链反应（RT-PCR）实验设计证实，每个新的DWV引物组对回顾性主变体具有特异性。ABC检测试剂盒的灵敏度取决于是否使用DNA或RNA作为模板，以及样本中是否存在其他主变体。通过下一代测序（NGS）数据确认，比较已知多样性样本中各主变体的总体比例，验证了ABC检测试剂盒的有效性。ABC测定法用于2006-2007年德文郡越冬菌落损失（OCL）研究的存档材料;进一步暗示DWV A型，并首次可能导致蜜蜂菌落过早崩溃。此外，在这项研究中，B型DWV与OCL无关。ABC检测的使用将使研究人员能够快速且经济有效地预筛选蜜蜂中DWV主变体的存在。
Deformed wing virus (DWV) is one of the most prevalent honey bee viral pathogens in the world. Typical of many RNA viruses, DWV is a quasi-species, which is comprised of a large number of different variants, currently consisting of three master variants: Type A, B, and C. Little is known about the impact of each variant or combinations of variants upon the biology of individual hosts. Therefore, we have developed a new set of master variant-specific DWV primers and a set of standards that allow for the quantification of each of the master variants. Competitive reverse transcriptase polymerase chain reaction (RT-PCR) experimental design confirms that each new DWV primer set is specific to the retrospective master variant. The sensitivity of the ABC assay is dependent on whether DNA or RNA is used as the template and whether other master variants are present in the sample. Comparison of the overall proportions of each master variant within a sample of known diversity, as confirmed by next-generation sequence (NGS) data, validates the efficiency of the ABC assay. The ABC assay was used on archived material from a Devon overwintering colony loss (OCL) 2006–2007 study; further implicating DWV type A and, for the first time, possibly C in the untimely collapse of honey bee colonies. Moreover, in this study DWV type B was not associated with OCL. The use of the ABC assay will allow researchers to quickly and cost effectively pre-screen for the presence of DWV master variants in honey bees.
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影响因子：
3
作者：
Brettell LE;Mordecai GJ;Schroeder DC;Jones IM;da Silva JR;Vicente-Rubiano M;Martin SJ
通讯作者：
Martin SJ
影响因子：
2.2
作者：
Genersch, E
通讯作者：
Genersch, E
影响因子：
1.9
作者：
Carreck, Norman L.;Bell, Brenda V.;Martin, Stephen J.
通讯作者：
Martin, Stephen J.
影响因子：
4.4
作者：
Dainat, Benjamin;Evans, Jay D.;Neumann, Peter
通讯作者：
Neumann, Peter
影响因子：
6.7
作者：
Bull JC;Ryabov EV;Prince G;Mead A;Zhang C;Baxter LA;Pell JK;Osborne JL;Chandler D
通讯作者：
Chandler D