The specific identification of animals through the analysis of faecal DNA is important in many areas of scientific endeavour, particularly in the field of parasitology. Here, we designed and assessed two multiplex PCR assays using genetic markers in a mitochondrial cytochrome b (cytb) gene region for the unequivocal identification and discrimination of animal species based on the specific amplification of DNA from faecal samples collected from water catchment areas in Victoria, Australia. One of these assays differentiates three marsupial species (eastern grey kangaroo, swamp wallaby and common wombat) and the other distinguishes three deer species (fallow, red and sambar deer). We tested these two assays using a total of 669 faecal samples, collected as part of an ongoing programme to monitor parasites and microorganisms in these animals. These two PCR assays are entirely specific for these animal species and achieve analytical sensitivities of 0.1–1.0 picogram (pg). We tested 669 faecal samples and found that some previous inferences of species based on faecal morphology were erroneous. We were able to molecularly authenticate all of the 669 samples. We have established PCR assays that accurately distinguish the faecal samples of some of the prominent large mammalian herbivores found within a water catchment system in the state of Victoria, Australia. The multiplex assays for marsupials and deer produce amplicons that are easily differentiable based on their size on an agarose gel, and can be readily sequenced for definitive species authentication. Although established for marsupials and deer, the methodology used here can be applied to other host-parasite study systems to ensure data integrity.

中文翻译：

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多重PCR用于从粪便样品中特异性鉴定有袋动物和鹿物种，作为寄生虫无创流行病学研究的基础

通过粪便DNA的分析对动物进行特异性鉴定在许多科学工作领域，尤其是在寄生虫学领域，都很重要。在这里，我们设计并评估了两种多重PCR检测方法，它们是利用线粒体细胞色素b（cytb）基因区域中的遗传标记对动物物种进行明确鉴定和区分的，基于从维多利亚州集水区收集的粪便样品中DNA的特异性扩增，澳大利亚。这些测定法之一可以区分三种有袋动物（东部灰袋鼠，沼泽鼠和普通袋熊），另一种可以区分三种鹿（小鹿，红鹿和水鹿）。我们使用总共669个粪便样品测试了这两种测定方法，这些样品是作为一项正在进行的计划的一部分而进行的，以监测这些动物的寄生虫和微生物。这两种PCR测定法完全针对这些动物物种，并且分析灵敏度为0.1–1.0皮克（pg）。我们测试了669个粪便样本，发现以前基于粪便形态学的物种推论是错误的。我们能够对669个样品进行分子鉴定。我们已经建立了可精确区分澳大利亚维多利亚州集水系统内发现的一些大型哺乳动物食草动物粪便样本的PCR分析方法。有袋动物和鹿的多重分析产生的扩增子基于琼脂糖凝胶上的大小容易区分，并且可以容易地测序以进行确定的物种鉴定。尽管为有袋动物和鹿而建立，