Hygienic monitoring of laboratory rodents has focused more and more on the analysis of environmental sample material by quantitative polymerase chain reaction (qPCR) assays. This approach requires profound knowledge of specific genetic sequences of the agents to be monitored and the assays need to be permanently adapted to take the latest research into account. [Pasteurella] pneumotropica was recently reclassified into the new genus Rodentibacter, with Rodentibacter (R.) pneumotropicus and R. heylii as the most commonly detected species in laboratory mouse colonies. This study aimed at the development of a specific qPCR assay for the simultaneous detection of both agents. A novel primer probe set, based on detection of the specific virulence factor' 'inclusion body protein A' gene (ibpA), was confirmed by testing the assay on currently described Rodentibacter type species and other Pasteurellaceae. Furthermore, it was validated within four different barrier units and results were compared with the cultural analysis of sentinel mice. The assay was suitable to specifically detect R. pneumotropicus and R. heylii and discriminate them from other murine Rodentibacter spp. In addition, it revealed high sensitivity for the detection of both agents in environmental sampling material including exhaust air dust in individually ventilated cage systems. Altogether, higher pathogen prevalence was detected via qPCR of environmental samples compared with cultural diagnostics of sentinel mice. This study describes a qPCR assay for the simultaneous detection of R. pneumotropicus and R. heylii. This assay was demonstrated to be beneficial during routine health monitoring, especially with regard to environmental sampling strategies.

中文翻译：

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一对二：一种新颖且高度灵敏的基于毒力因子的定量聚合酶链反应测定法，用于同时检测环境样品材料中的嗜气性杆状杆菌和海氏杆状杆菌。

实验室啮齿动物的卫生监测越来越关注通过定量聚合酶链反应（qPCR）分析对环境样品材料的分析。这种方法需要对要监测的试剂的特定遗传序列有深入的了解，并且需要永久调整分析方法以考虑最新的研究。[Pasteurella] pneumotropica最近被重新归类为新的Rodentibacter属，其中Rodentibacter（r。）pneumotropicus和R. heylii是实验室小鼠菌落中最常见的物种。这项研究旨在开发一种同时检测两种药物的特异性qPCR分析方法。一种新颖的引物探针组，基于对特定毒力因子“包涵体蛋白A”基因（ibpA）的检测，通过对当前描述的杆状杆菌属物种和其他巴斯德杆菌科的检测方法进行了测定，结果得到了证实。此外，它在四个不同的屏障单元内得到验证，并将结果与​​前哨小鼠的文化分析进行了比较。该测定法适合于特异性检测嗜肺性罗勒氏菌和海伊氏菌，并将它们与其他鼠类杆状杆菌属分离。此外，它还显示出对环境采样材料中两种试剂（包括在单独通风的笼式系统中的废气灰尘）进行检测的高灵敏度。总的来说，与前哨小鼠的文化诊断相比，通过环境样品的qPCR检测到更高的病原体患病率。这项研究描述了同时检测R. pneumotropicus和R. heylii的qPCR分析。