不同材质的微生物捕捉滤膜在屏障环境设施中的应用效果初探  

Exploration on Application Effectiveness of Microbial Capture Filter Membranes from Different Materials in Barrier Environment Facilities

作　　者：孔垂琴 田苗苗 蔡利东 吴东 倪露 张楚漫 杨慧欣 KONG Chuiqin;TIAN Miaomiao;CAI Lidong;WU Dong;NI Lu;ZHANG Chuman;YANG Huixin(GemPharmatech Co.,Ltd.,Nanjing 210032,China)

机构地区：[1]江苏集萃药康生物科技股份有限公司,南京210032

出　　处：《实验动物与比较医学》2024年第4期436-444,共9页Laboratory Animal and Comparative Medicine

基　　金：江苏省实验动物协会2023年研究课题“笼架排风粉尘捕捉器滤材材料探索及多重EAD-QPCR检测方法建立”(DWXH202305)。

摘　　要：目的通过探究不同材质滤膜的微生物捕捉性能,评估其对屏障环境实验设施中啮齿类动物健康状况的监测效果。方法以嗜肺巴斯德杆菌和金黄色葡萄球菌为代表菌株,在实验室条件下模拟滤膜捕捉微生物的过程,以捕捉粉尘质量、最低检测限度及荧光定量PCR检测的Ct值差异综合评估5种具有吸附、透气性能的自选材质滤膜(M1~M5)和商品化捕捉滤膜(T1)的微生物捕捉效果。将性能最优的自选材质滤膜置于屏障设施内笼架的通风管道中,用相应笼架的哨兵鼠作为对照组,以表皮葡萄球菌和大肠埃希菌为指示菌,计算阳性检出率和符合率,探讨其作为微生物捕捉滤膜在屏障设施内监测实验动物健康状况的可行性。结果从捕捉粉尘质量看,自选材质滤膜M3(滤径为0.1µm的无纺布可作为材质)的捕捉效果仅次于T1,捕捉量达0.126 g;对于金黄色葡萄球菌,除M4外,其余材质的滤膜最低检测限度均为102 CFU/g;对于嗜肺巴斯德杆菌,所有滤膜的最低检测限度皆为102 CFU/g;但M3的荧光定量PCR扩增结果Ct值显著小于其他材质,故5种自选材质滤膜中M3的捕捉效果最好。滤膜检测验证实验中,表皮葡萄球菌在哨兵鼠粪便和M3中的阳性检出率分别为50.00%(6/12)和58.33%(7/12),符合率为92%;大肠埃希菌在哨兵鼠粪便和M3中阳性检出率均为50.00%(6/12),符合率为100%。结论5种自选材质滤膜中,M3捕捉效果最佳;在屏障环境设施内,M3对于表皮葡萄球菌的监测效果优于哨兵鼠。因此,滤径为0.1µm的无纺布可作为微生物捕捉滤膜材质,为笼架排风粉尘PCR监测用微生物捕捉滤膜的材质选择和应用提供借鉴经验。Objective To evaluate the effectiveness of filter membranes made from different materials in monitoring the health status of rodents in barrier environment facilities by investigating their microbial capture performance.Methods Pasteurella pneumotropica(Pp)and Staphylococcus aureus(Sa)were used as representative strains to simulate the process of microbial capture by filter membranes under laboratory conditions.The microbial capture effectiveness of five self-selected filter membranes(M1,M2,M3,M4,and M5)with adsorption and breathability properties and a commercial filter membrane(T1)were comprehensively evaluated based on captured dust mass,minimum detection limit,and differences in Ct values obtained through fluorescence quantitative PCR detection.The best-performing self-selected filter membrane was placed in the ventilation ducts of cage racks within the barrier facility,with sentinel mice in corresponding cage racks as the control group.Staphylococcus epidermidis and Escherichia coli were used as indicator bacteria to calculate the positive detection rate and coincidence rate,thereby exploring the feasibility of using microbial capture filter membranes to monitor the health status of experimental animals in barrier facilities.Results In terms of the captured dust mass,the self-selected filter membrane M3(non-woven filter membrane with a diameter of 0.1µm);showed a capture effectiveness second only to T1,with a capture mass of 0.126 g.For Sa,all filter membranes except M4 had a minimum detection limit of 10²CFU/g.For Pp,the minimum detection limit for all filter membranes was 102 CFU/g.However,the Ct value of the quantitative fluorescence PCR amplification results for M3 was significantly lower than that of other materials,indicating that M3 had the best capture performance among the five self-selected materials.In the filter detection verification experiment,the positive detection rate of Staphylococcus epidermidis in sentinel mouse feces and M3 was 50.00%(6/12)and 58.33%(7/12),respectively,with a coinciden

关 键 词：微生物捕捉滤膜 屏障环境设施 笼架排风粉尘 荧光定量PCR法 哨兵鼠 

分 类 号：Q95-33[生物学—动物学] R-332[医药卫生]