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通过使用经聚合酶链反应(PCR)验证的汽化过氧化氢在整个设施范围内进行根除。

Facility-wide Eradication of by using PCR-validated Vaporized Hydrogen Peroxide.

作者信息

Miedel Emily L, Ragland Natalie H, Engelman Robert W

机构信息

Comparative Medicine, H. Lee Cancer Center & Research Institute, University of South Florida, Tampa, Florida;, Email:

Comparative Medicine, H. Lee Cancer Center & Research Institute, University of South Florida, Tampa, Florida.

出版信息

J Am Assoc Lab Anim Sci. 2018 Sep 1;57(5):465-476. doi: 10.30802/AALAS-JAALAS-17-000135. Epub 2018 Jul 13.

Abstract

Facility-wide Corynebacterium bovis eradication was established using vaporized hydrogen peroxide (VHP) decontamination guided by C. bovis PCR surveillance. Prior attempts limited to culling PCR-positive mice and decontaminating affected rooms were ineffective in preventing recurrence. Because research aims often require trafficking to and use of procedural cores, a 12-mo facility-wide C. bovis PCR surveillance of 2064 specimens was performed and documented that, despite the presence of few clinically hyperkeratotic mice, 35% of the murine housing and use space was contaminated by C. bovis. The airways of IVC racks and air-handling units (AHU) provided a substantive niche for C. bovis survival, comparable to the primary enclosure, with 26% of murine and 22% of airway specimens PCR-positive for C. bovis. Equipment airway VHP sterilization in a 'flex room' required an 'active-closed' setting with the IVC rack connected to the AHU set to the VHP cycle, because 12% of specimens from 'static-open' VHP-exposed airways remained PCR-positive for C. bovis, whereas 0% of specimens from active-closed VHP exposures were positive. VHP decontamination of the 29,931-ft2 facility was completed in 2 mo. C. bovis PCR testing of IVC exhaust plenums for 200 d in previously C. bovis-affected rooms confirmed that none of the 259 specimens tested were PCR-positive for the organism. Monthly surveillance identified a single recurrence during June 2017 (month 9), ensuring rapid culling of C. bovis PCR-positive mice and acute VHP decontamination of equipment and rooms. Molecular persistence of C. bovis was resolved in procedural and personnel areas, and no murine or housing specimens tested C. bovis PCR-positive during study months 11 and 12. Furthermore, since the conclusion of the 12-mo study, none of the 452 additional murine, cell biologic, environmental, and monthly equipment surveillance specimens tested were C. bovis PCR-positive, documenting an 11-mo period of facility-wide C. bovis eradication to date. Study invalidation due to C. bovis can be avoided through PCR surveillance for the organism, immediate culling of PCR-positive mice, and acute VHP decontamination of affected areas.

摘要

在牛棒状杆菌聚合酶链反应(PCR)监测的指导下,使用汽化过氧化氢(VHP)去污法在整个设施范围内根除牛棒状杆菌。之前仅限于扑杀PCR检测呈阳性的小鼠并对受影响的房间进行去污的尝试,在预防复发方面没有效果。由于研究目的通常需要将动物转运至程序核心区域并在其中使用,因此对2064份样本进行了为期12个月的全设施范围牛棒状杆菌PCR监测,结果表明,尽管临床上只有少数小鼠出现角化过度,但35%的小鼠饲养和使用空间受到牛棒状杆菌污染。独立通风笼具(IVC)架和空气处理单元(AHU)的气道为牛棒状杆菌的存活提供了一个重要的生态位,与主要饲养环境相当,26%的小鼠样本和22%的气道样本牛棒状杆菌PCR检测呈阳性。在“灵活空间”对设备气道进行VHP消毒时,需要设置“主动-关闭”模式,即将IVC架连接到AHU并设置为VHP循环模式,因为在“静态-开放”VHP暴露气道的样本中,12%的样本牛棒状杆菌PCR检测仍呈阳性,而在主动-关闭VHP暴露的样本中,这一比例为0%。对面积为29,931平方英尺的设施进行VHP去污在2个月内完成。对之前受牛棒状杆菌影响的房间的IVC排气增压室进行了200天的牛棒状杆菌PCR检测,结果证实,所检测的259份样本中没有一份该生物体PCR检测呈阳性。每月监测发现2017年6月(第9个月)出现一次复发情况,随即迅速扑杀牛棒状杆菌PCR检测呈阳性的小鼠,并对设备和房间进行急性VHP去污。牛棒状杆菌在程序和人员区域的分子残留问题得到解决,在研究的第11个月和第12个月,没有小鼠或饲养样本牛棒状杆菌PCR检测呈阳性。此外,自为期12个月的研究结束以来,另外检测的452份小鼠、细胞生物学、环境和每月设备监测样本中,没有一份牛棒状杆菌PCR检测呈阳性,这表明到目前为止,该设施范围内已实现了为期11个月的牛棒状杆菌根除。通过对该生物体进行PCR监测、立即扑杀PCR检测呈阳性的小鼠以及对受影响区域进行急性VHP去污,可以避免因牛棒状杆菌导致的研究无效。

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