Amdoparvoviruses infect carnivore species, including mink, raccoon dog, fox, skunk, and red panda. Amdoparvovirus infection is a major cause of morbidity and mortality in farmed minks. Here, we developed a direct TaqMan qPCR assay for detection and quantification of carnivore amdoparvoviruses by using three primers and one probe based on the conserved VP2 gene. The detection limit for Aleutian mink disease virus (AMDV) and Raccoon dog and arctic fox amdoparvovirus (RFAV) were 4.06 × 101 copies/μl and 2.93 × 101 copies/μl, respectively. Both intra- and inter-assay variability were less than 2%. Among 74 carnivore samples, the positive rates for amdoparvoviruses were 62.2% (46/74) by direct TaqMan qPCR, while only 40.5% (30/74) by SYBR Green I qPCR. This result suggests that the direct TaqMan qPCR was more sensitive than the SYBR Green I qPCR. Additionally, the direct TaqMan qPCR is a rapid and sensitive method for liquid samples at microliter level as the assay employed the direct alkaline lysis method to obtain viral DNA and, therefore, eliminated the cumbersome steps in extracting DNA. Overall, the direct TaqMan qPCR assay possessed high specificity, sensitivity, and reproducibility, indicating that it can be used as a powerful tool for detection and quantification of various carnivore amdoparvoviruses in epidemiological and pathogenesis studies.

中文翻译：

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开发和评估直接TaqMan qPCR测定法以快速检测各种食肉动物amdoparvoviruses。

Amdoparvoviruses病毒感染肉食动物，包括貂、,、狐狸，臭鼬和小熊猫。Amdoparvovirus感染是养殖水貂发病和死亡的主要原因。在这里，我们开发了一种直接的TaqMan qPCR检测方法，通过使用三个基于保守VP2基因的引物和一个探针来检测和定量肉食性双歧细小病毒。阿留申水貂病病毒（AMDV），R狗和北极狐细小病毒（RFAV）的检出限分别为4.06×101拷贝/μl和2.93×101拷贝/μl。批内和批间变异均小于2％。在74个食肉动物样本中，通过直接TaqMan qPCR检测到的amdoparvovirus病毒的阳性率为62.2％（46/74），而通过SYBR Green I qPCR检测的仅40.5％（30/74）。该结果表明直接TaqMan qPCR比SYBR Green I qPCR更为灵敏。此外，直接TaqMan qPCR是一种用于微升水平液体样品的快速灵敏方法，因为该测定法采用直接碱裂解法获得病毒DNA，因此省去了提取DNA的繁琐步骤。总体而言，直接TaqMan qPCR检测具有很高的特异性，敏感性和可重复性，表明它可以用作流行病学和发病机制研究中检测和定量检测各种食肉动物amdoparvoviruses的有力工具。