Department of Comparative Medicine, University of Washington, Seattle, Washington;, Email:
Department of Comparative Medicine, University of Washington, Seattle, Washington.
J Am Assoc Lab Anim Sci. 2020 Mar 1;59(2):156-162. doi: 10.30802/AALAS-JAALAS-19-000079. Epub 2020 Feb 19.
Reliable detection of unwanted microbial agents is essential for meaningful health monitoring in laboratory animal facilities. Most rodents at our institution are housed in IVC rack systems to minimize aerogenic transmission of infectious agents between cages. The most commonly used rodent health monitoring systems expose live sentinel rodents to soiled bedding collected from other rodent cages on IVC racks and subsequently test these soiled-bedding sentinels for evidence of infection with excluded agents. However, infectious agents might go undetected when using this health surveillance method, due to inefficient organism shedding or transmission failure. In 2016, our institution switched the health monitoring methodology for the majority of our SPF rodent colonies to real-time PCR testing of environmental samples collected from the exhaust plenums of IVC racks. Here we describe our rationale for this conversion, describe some interesting health monitoring cases that arose soon after the conversion, and discuss a potential problem with the conversion-residual nucleic acids. We compared cost and implementation effort associated with 2 sampling methods, sticky swabs and in-line collection media. We also compared the ability of these 2 sampling methods to detect 2 prevalent microbes in our facilities, and murine norovirus. Our institution-wide switch to health monitoring by real-time PCR assay of exhaust air dust samples thus far has provided a sensitive, simple, and reliable approach for maintenance of SPF conditions in laboratory rodents and has dramatically reduced the use of live sentinel animals.
可靠地检测到不需要的微生物对于实验室动物设施中有意义的健康监测至关重要。我们机构的大多数啮齿动物都被安置在 IVC 架系统中,以最大限度地减少笼与笼之间空气传播的传染性病原体。最常用的啮齿动物健康监测系统将活的监测动物暴露于从 IVC 架上的其他啮齿动物笼中收集的污染垫料中,然后对这些污染垫料监测动物进行检测,以发现排除的病原体感染的证据。然而,使用这种健康监测方法时,由于生物体的低效脱落或传播失败,可能会检测不到传染性病原体。2016 年,我们机构将大多数 SPF 啮齿动物群的健康监测方法改为从 IVC 架的排气通风孔中收集的环境样本的实时 PCR 检测。在这里,我们描述了我们转换的基本原理,描述了转换后不久出现的一些有趣的健康监测案例,并讨论了转换后的潜在问题——残留核酸。我们比较了两种采样方法(粘性拭子和在线收集介质)的成本和实施工作量。我们还比较了这两种采样方法检测我们设施中两种常见微生物(鼠诺如病毒)的能力。我们机构范围内通过实时 PCR 检测排气粉尘样本进行健康监测的转换,迄今为止为维持实验动物 SPF 条件提供了一种敏感、简单和可靠的方法,并大大减少了活监测动物的使用。