EcoGene®, Landcare Research, Auckland, New Zealand.
Electrophoresis. 2013 Dec;34(24):3370-6. doi: 10.1002/elps.201300324. Epub 2013 Nov 13.
An identification assay has been developed that allows accurate detection of 19 of the most common terrestrial mammals present in New Zealand (cow, red deer, goat, dog, horse, hedgehog, cat, tammar wallaby, mouse, weasel, ferret, stoat, sheep, rabbit, Pacific rat, Norway rat, ship rat, pig, and brushtail possum). This technique utilizes species-specific primers that, combined in a multiplex PCR, target small fragments of the mitochondrial cytochrome b gene. Each species, except hedgehog, produces two distinctive species-specific fragments, making the assay self-confirmatory and enabling the identification of multiple species simultaneously in DNA mixtures. The multiplex assay detects as little as 100 copies of mitochondrial DNA, which makes it a very reliable tool for degraded and trace samples. Reliability, accuracy, reproducibility, and sensitivity tests to validate the technique were performed. The technique featured here enabled a prompt response in a predation specific event, but can also be useful for wildlife management and conservation, pest incursions detection, forensic, and industrial purposes in a very simple and cost-effective manner.
已经开发出一种鉴定检测方法，可准确检测到新西兰现存的 19 种最常见的陆地哺乳动物（奶牛、马鹿、山羊、狗、马、刺猬、猫、塔马尔沙袋鼠、老鼠、黄鼠狼、雪貂、白鼬、绵羊、兔子、太平洋鼠、挪威鼠、褐家鼠、猪和帚尾袋貂）。该技术利用物种特异性引物，在多重 PCR 中靶向线粒体细胞色素 b 基因的小片段。除刺猬外，每个物种都会产生两个独特的物种特异性片段，使该检测具有自我确认性，并能够同时在 DNA 混合物中鉴定多种物种。该多重检测方法可检测低至 100 个线粒体 DNA 拷贝，这使其成为降解和痕量样本的非常可靠工具。为验证该技术进行了可靠性、准确性、可重复性和灵敏度测试。这里介绍的技术能够在特定捕食事件中做出快速响应，但也可以非常简单且具有成本效益的方式用于野生动物管理和保护、害虫入侵检测、法医和工业用途。
