啮齿类实验动物健康监测用脏垫料哨兵动物法和排风粉尘PCR法比较

doi:10.12300/j.issn.1674-5817.2023.168

实验动物与比较医学 ›› 2024, Vol. 44 ›› Issue (3): 321-327.DOI: 10.12300/j.issn.1674-5817.2023.168

啮齿类实验动物健康监测用脏垫料哨兵动物法和排风粉尘PCR法比较

于灵芝¹, 魏晓锋¹()(), 黎明², 孔志豪¹

^1.上海实验动物研究中心, 上海 201203
^2.英崴沃(上海)生物科技有限公司, 上海 201203

收稿日期:2023-11-22 修回日期:2024-01-30 出版日期:2024-07-06 发布日期:2024-06-25
通讯作者: 魏晓锋（1980—），男，硕士，副研究员，主要从事实验动物质量控制研究。E-mail： wei.xf@outlook.com。ORCID： 0009-0009-5089-8342
作者简介:于灵芝（1980—），女，博士，助理研究员，主要从事病原体核酸检测方法研究。E-mail： yulingzhi@slarc.org.cn
基金资助:
上海实验动物研究中心科技创新计划新星项目“木糖葡萄球菌快速分子检测方法的建立及应用”(2024NS05);上海市科技计划项目资助“上海市水生实验动物专业技术服务平台”(22DZ2291200)

Comparison of Methods between Soiled Bedding Sentinels and Exhaust Air Dust PCR for Health Monitoring of Rodent Laboratory Animals

Lingzhi YU¹, Xiaofeng WEI¹()(), Ming LI², Zhihao KONG¹

^1.Shanghai Laboratory Animal Research Center, Shanghai 201203, China
^2.INVIVO (Shanghai) Biotechnology Co. , Ltd. , Shanghai 201203, China

Received:2023-11-22 Revised:2024-01-30 Published:2024-06-25 Online:2024-07-06
Contact: WEI Xiaofeng (ORCID: 0009-0009-5089-8342), E-mail: wei.xf@outlook.com

摘要/Abstract

摘要：

实验动物微生物质量对科学研究数据的有效性和重复性以及人类健康和动物福利至关重要。目前，独立通风笼具（individual ventilation cage，IVC）已成为主流的啮齿类实验动物饲养系统。针对这种饲养方式的病原体监测方法最常用的是脏垫料哨兵动物法（soiled bedding sentinels，SBS），该方法是以间接接触和延迟反馈的方式监测鼠群的微生物携带状况，能有效监测通过粪-口途径传播的病原体如小鼠肝炎病毒、呼肠孤病毒等。但这种方法难以监测到主要通过气溶胶、直接接触等途径传播的病原体，例如仙台病毒、嗜肺巴斯德杆菌等。排风粉尘（exhaust air dust，EAD）-PCR监测方法分为在IVC笼架排风管道中拭子采样，用以监测管道相对应的笼架；主机初效过滤前拭子采样，用以监测整个IVC笼架；EAD收集装置采样，用以监测同一个主机连接的所有笼架。不同IVC厂商针对各自的IVC系统开发了相应的EAD收集装置，使操作便捷，容易实现标准化。目前，与SBS方法相比，EAD-PCR方法的检出率和时效性都有显著提升，最快暴露一周就可检出，可作为SBS方法的补充或替代，有利于维护实验动物福利。本文对上述两种病原体监测方法的应用进展进行综述，同时结合本实验室和送检单位EAD-PCR监测的实施情况，对该方法存在的局限性进行分析并提出解决方案。EAD-PCR方法有助于减少活体哨兵动物的使用量，可以更好地维护实验动物福利的“3Rs”原则。

关键词: 啮齿类实验动物, 健康监测, 脏垫料哨兵, 排风粉尘PCR监测

Abstract:

The microbiological quality of laboratory animals is crucial for the validity and reproducibility of scientific research data, as well as human health and animal welfare. Currently, individual ventilation cages (IVC) have become the mainstream feeding system for rodent laboratory animals. The most commonly used pathogen monitoring method for this feeding system is soiled bedding sentinels (SBS). This method monitors the microbial carrying status of mouse colony through indirect contact and delayed feedback. It can effectively monitor pathogens transmitted via the fecal-oral route, such as mouse hepatitis virus and reovirus. However, this method has difficulty detecting pathogens mainly transmitted through aerosols or direct contact, such as Sendai virus and Pasteurella pneumotropica. The exhaust air dust (EAD)-PCR monitoring method involves swab sampling in the IVC exhaust ducts to monitor the corresponding racks of the ducts; swab sampling before the prefiltration of the host to monitor the entire IVC rack; and EAD collection device sampling to monitor all racks connected to the same host. Different IVC manufacturers have developed corresponding EAD collection devices for their respective IVC systems, making operations convenient and standardization easy. Compared with the SBS method, the EAD-PCR method significantly improves detection rate and timeliness, with the fastest detection possible after one week of exposure. It can serve as a supplement or replacement for the SBS method. Currently, increasing evidence supports that EAD-PCR testing is a more reliable, sensitive, and cost-effective monitoring method, and is more beneficial to animal welfare. This article reviews the application progress of these two methods for monitoring pathogens, analyzes the existing limitations of the EAD-PCR method, and proposes solutions based on its implementation in our laboratory and examination units. The EAD-PCR method helps reduce the number of live sentinel animals used in pathogen monitoring, in order to better maintain the "3Rs" principle of laboratory animal welfare.

Key words: Rodent laboratory animals, Health monitoring, Soiled bedding sentinels, Exhaust air dust- PCR monitoring

中图分类号:

Q95-33

于灵芝,魏晓锋,黎明,等. 啮齿类实验动物健康监测用脏垫料哨兵动物法和排风粉尘PCR法比较[J]. 实验动物与比较医学, 2024, 44(3): 321-327. DOI: 10.12300/j.issn.1674-5817.2023.168.

Lingzhi YU,Xiaofeng WEI,Ming LI,et al. Comparison of Methods between Soiled Bedding Sentinels and Exhaust Air Dust PCR for Health Monitoring of Rodent Laboratory Animals[J]. Laboratory Animal and Comparative Medicine, 2024, 44(3): 321-327. DOI: 10.12300/j.issn.1674-5817.2023.168.

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啮齿类实验动物健康监测用脏垫料哨兵动物法和排风粉尘PCR法比较

Comparison of Methods between Soiled Bedding Sentinels and Exhaust Air Dust PCR for Health Monitoring of Rodent Laboratory Animals

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