Quantitative PCR multiplexes for simultaneous multispecies detection of Asian carp eDNA
Quantitative PCR multiplexes for simultaneous multispecies detection of Asian carp eDNA
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DOI：
10.1016/j.jglr.2017.05.001
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发表时间：
2017-08
影响因子：
2.2
通讯作者：
K. Wozney;C. Wilson
中科院分区：
文献类型：
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作者：
K. Wozney;C. Wilson
环境DNA（eDNA）越来越多地被用作检测水生入侵物种的监测工具。特别是，五大湖周围的许多管辖区正在采用这种方法来监测亚洲鲤鱼的潜在存在。我们开发了两种多重定量PCR（qPCR）反应，用于同时检测四种亚洲鲤鱼的eDNA，这些鲤鱼已被确定为五大湖的潜在入侵者[大头鱼（Hypophthalmichthys nobilis）、银鲤（Hypophthalmichthys molitrix）、青鱼（青鱼）和草鱼（草鱼）]，沿着内部对照，以测试由于样品污染引起的假阳性结果。这些检测方法具有高度灵敏度，能够在实验室对照中检测出微量的鲤鱼DNA。多重设计的使用允许同时测试多个物种，减少加工和处理时间，降低成本并减少测试所需的DNA提取物的量。在这项研究中描述的多重检测可以很容易地应用到亚洲鲤鱼监测计划，目前正在使用的支持监测工作在加拿大沃茨的伊利湖，休伦湖和安大略湖的所有四种亚洲鲤鱼。
Environmental DNA (eDNA) is increasingly being used as a surveillance tool for detection of aquatic invasive species. In particular, this method is being applied by many jurisdictions surrounding the Great Lakes to monitor for the potential presence of Asian carp. We developed two multiplex quantitative PCR (qPCR) reactions for the simultaneous testing for eDNA from the four species of Asian carp that have been identified as potential invaders into the Great Lakes [bighead carp (Hypophthalmichthys nobilis), silver carp (Hypophthalmichthys molitrix), black carp (Mylopharyngodon piceus), and grass carp (Ctenopharyngodon idella)], along with internal controls to test for false positive results due to sample contamination. The assays are highly sensitive and have the ability to detect trace amounts of carp DNA in laboratory controls. The use of a multiplex design allows for simultaneous testing for numerous species, reducing processing and handling times, lowering costs and reducing the amount of DNA extract required for testing. The multiplex assays described in this study may be readily applied to Asian carp monitoring programs, and are currently being used to support surveillance efforts in the Canadian waters of Lake Erie, Lake Huron and Lake Ontario for all four species of Asian carp.