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检测独立通风笼具中的鼠星状病毒和禽螨:探索适合日常卫生监测的检测方法。

Detection of Murine Astrovirus and Myocoptes musculinus in individually ventilated caging systems: Investigations to expose suitable detection methods for routine hygienic monitoring.

机构信息

Helmholtz Zentrum München-German Research Center for Environmental Health, Research Unit Comparative Medicine, Neuherberg, Germany.

出版信息

PLoS One. 2019 Aug 13;14(8):e0221118. doi: 10.1371/journal.pone.0221118. eCollection 2019.

Abstract

Murine Astrovirus is one of the most prevalent viral agents in laboratory rodent facilities worldwide, but its influence on biomedical research results is poorly examined. Due to possible influence on research results and high seroprevalence rates in mice, it appears useful to include this virus into routine health monitoring programs. In order to establish exhaust air particle PCR as a reliable detection method for Murine Astrovirus infections in mice kept in individually ventilated cages (IVC) and compare the method to sentinel mice monitoring regarding reproducibility and detection limit, we conducted a study with defined Murine Astrovirus cage prevalence. In parallel, the efficacy of both detection strategies (soiled-bedding sentinel (SBS) and exhaust air dust (EAD) analysis) was tested for Myocoptes musculinus. The fur mite was used as a reference organism during the whole study period to ensure the validity of this method. Because some publications already demonstrated successful detection of several pathogens, including murine fur mite species, via EAP-PCR. Detection of Murine Astrovirus infections at low prevalence is possible with both methods tested. Detection by exhaust air particles (EAP) is faster, more sensitive and more reliable compared to soiled bedding sentinels (SBS). Exhaust air particle PCR also detected the reference organism Myocoptes musculinus, which was not detected at all by sentinel mice, not even by high sensitivity fur swab qPCR. In conclusion, Murine Astrovirus can be detected by both exhaust air particle PCR and soiled bedding sentinels. We recommend exhaust air particle PCR as the better detection technique for Murine Astrovirus, because it is more reliable. Environmental samples are the method of choice for detection of Myocoptes musculinus because relying on soiled bedding sentinels harbors a big risk of missing existing infestations.

摘要

鼠星状病毒是世界范围内实验室啮齿动物设施中最常见的病毒之一,但它对生物医学研究结果的影响尚未得到充分研究。由于该病毒可能会影响研究结果,并且在小鼠中的血清阳性率较高,因此似乎有必要将其纳入常规健康监测计划中。为了建立废气颗粒 PCR 作为一种可靠的检测方法,用于检测在独立通风笼(IVC)中饲养的小鼠中的鼠星状病毒感染,并比较该方法与哨兵鼠监测的重现性和检测限,我们进行了一项具有明确鼠星状病毒笼流行率的研究。同时,还测试了两种检测策略(污染垫哨兵(SBS)和废气尘埃(EAD)分析)对肌鼠螨的效果。在整个研究期间,毛螨都被用作参考生物,以确保该方法的有效性。因为一些出版物已经证明,通过 EAP-PCR 可以成功检测到包括鼠螨物种在内的多种病原体。通过两种方法均可以检测到低流行率的鼠星状病毒感染。与污染垫哨兵(SBS)相比,废气颗粒(EAP)的检测速度更快、更敏感且更可靠。废气颗粒 PCR 还检测到了参考生物肌鼠螨,而哨兵鼠根本无法检测到该生物,甚至使用高灵敏度的毛拭子 qPCR 也无法检测到。总之,鼠星状病毒可以通过废气颗粒 PCR 和污染垫哨兵检测到。我们建议使用废气颗粒 PCR 作为检测鼠星状病毒的更好的检测技术,因为它更可靠。环境样本是检测肌鼠螨的首选方法,因为依赖污染垫哨兵存在错过现有感染的巨大风险。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6836/6692027/ad6a28284aea/pone.0221118.g001.jpg

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