Comparison of Rodent Infectious Agent Detection by Exhaust Dust Testing and Traditional Sentinel Testing Using Quantitative Polymerase Chain Reaction.

Taylor Simmons, Yesen Zhou, Lea Ann Chlebek, Cherie Chang, Lexi Frank, Jason Villano, Cheryl Perkins, Ken Henderson, Zachary T Freeman
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Abstract

Improved diagnostic capabilities and a desire to reduce or refine the use of animals as soiled bedding sentinels (SBS) have driven interest in developing the use of PCR-based testing methods, such as exhaust dust testing (EDT), for routine rodent health surveillance. We compared the absolute and quantitative PCR results from EDT filters with SBS mice by routine screening via a panel of 19 infectious agents including agents known to be excluded or present in the colony. In this study, EDT and SBS were compared at days 0, 90, and 180 in 3 facilities (n = 12 rooms) with animals housed on IVC racks (n = 19 double-sided and 23 single-sided racks). All racks were negative for excluded agents (n = 15 agents) during the study. The bacterial agent Helicobacter spp. was consistently detected on EDT filters while less consistently detected via SBS. EDT filters detected Corynebacterium bovis better than SBS in areas where the agent was present. EDT filters and SBS mice tested for murine norovirus (MNV) demonstrated agreement for positive tests by both PCR and serology. For rodent chaphamaparvovirus-1 (RCHPV-1) we compared EDT to urine and feces from SBS. Six cages of SBS were positive for RCHPV-1 by fecal PCR with 5 out of 6 testing positive on urine, while only 3 out of 6 EDT filters tested positive. Since real-time fluorogenic PCR was used for testing, relative PCR copy numbers for each positive finding were evaluated to estimate organism load at the rack level. Copy numbers allowed for further characterization of agent presence within a colony. Furthermore, we compared copy numbers with cage census for MNV and Helicobacter spp., which was positively correlated for EDT testing but not for SBS. Overall, our results demonstrate that EDT's ability to detect many commonly excluded agents is comparable to or better than SBS.

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使用定量聚合酶链式反应进行排气粉尘检测和传统哨点检测的鼠类传染病病原体检测比较。
由于诊断能力的提高以及减少或改进使用动物作为污秽垫料哨兵(SBS)的愿望,人们对开发使用基于 PCR 的检测方法(如排气粉尘检测(EDT))进行常规啮齿动物健康监测产生了兴趣。我们比较了 EDT 过滤器与 SBS 小鼠的绝对和定量 PCR 结果,通过 19 种传染病病原体(包括已知排除或存在于啮齿动物群中的病原体)进行常规筛查。在这项研究中,在 3 个设施(n = 12 个房间)中对第 0、90 和 180 天的 EDT 和 SBS 进行了比较,动物饲养在 IVC 架上(n = 19 个双面架和 23 个单面架)。在研究期间,所有架子上的排除病原体(n = 15 种病原体)均为阴性。在 EDT 过滤器上持续检测到螺旋杆菌属细菌,而通过 SBS 则检测不到。在存在科里纳杆菌的区域,EDT 过滤器比 SBS 更能检测出科里纳杆菌。通过 PCR 和血清学方法检测小鼠诺瓦克病毒 (MNV) ,EDT 过滤器和 SBS 小鼠的阳性检测结果一致。对于啮齿动物瘤胃病毒-1(RCHPV-1),我们将 EDT 与 SBS 的尿液和粪便进行了比较。通过粪便 PCR,6 个 SBS 笼中有 5 个对 RCHPV-1 检测呈阳性,而 6 个 EDT 过滤器中只有 3 个对 RCHPV-1 检测呈阳性。由于检测使用的是实时荧光 PCR,因此评估了每个阳性结果的相对 PCR 拷贝数,以估计机架上的生物量。通过拷贝数可以进一步确定菌落中是否存在病原体。此外,我们还将拷贝数与笼中 MNV 和螺旋杆菌的普查结果进行了比较,发现 EDT 检测结果与笼中 MNV 和螺旋杆菌的普查结果呈正相关,而 SBS 检测结果与笼中 MNV 和螺旋杆菌的普查结果不呈正相关。总之,我们的结果表明,EDT 检测许多通常被排除在外的病原体的能力与 SBS 相当,甚至更好。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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American Society of Laboratory Animal Practitioners Position Statement: Handling and Physical Restraint of Research Animals. American Society of Laboratory Animal Practitioners Position Statement: Definition of Animal Welfare. Effect of Novel High-fat Diet Feeding Methods on Food Wastage, Weight Gain, Hair Coat Grease Accumulation, and Scratching Behavior in C57BL/6NCrl Mice. Identification and Treatment of Fur Mites (Radfordia lemnina) in California Deer Mice (Peromyscus californicus) Using Selamectin. American Society of Laboratory Animal Practitioners Position Statement: Animal Care Principles.
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