广西某高校细胞培养实验室污染状况的调查研究  

Investigation on the pollution of cell culture laboratory in a university in Guangxi

作　　者：钟伟月 黄海燕 冯燚 彭远军 廖艳研 王海龙 梁敏莉 林成琳 杨扬 叶力 潘沛江 梁浩 Zhong Weiyue;Huang Haiyan;Feng Yan;Peng Yuanjun;Liao Yanyan;Wang Hailong;Liang Minli;Lin Chenglin;Yang Yang;Ye Li;Pan Peijiang;Liang Hao(Guangxi Key Laboratory of AIDS Prevention and Control,School of Public Health,Guangxi Medical University,Nanning 530021,China;Biosafety Level 3 Laboratory,Life Sciences Institute of Guangxi Medical University,Nanning 530021,China;China(Guangxi)-ASEAN Joint Laboratory of Emerging Infectious Disease,Nanning 530021,China)

机构地区：[1]广西医科大学公共卫生学院广西艾滋病防治研究重点实验室,南宁530021 [2]广西医科大学生命科学研究院生物安全三级实验室,南宁530021 [3]中国(广西)-东盟新发传染病联合实验室,南宁530021

出　　处：《广西医科大学学报》2022年第9期1469-1473,共5页Journal of Guangxi Medical University

基　　金：广西医科大学2021年大学生创新创业训练计划项目(No.202110598293)。

摘　　要：目的:通过了解广西某高校细胞培养实验室的运行管理情况,以及检测细菌、支原体污染状况,为促进实验室的规范管理提供参考依据。方法:在广西某高校随机抽取8个细胞培养实验室并进行采样,琼脂细菌培养法检测细菌总数,PCR扩增及琼脂糖凝胶电泳检测支原体污染情况。结果:本研究所调查的实验室各采样点均存在不同程度的细菌污染和支原体污染,其中房间角落、地面中央、安全柜落脚处、培养箱外侧扶手以及内壁均可检出细菌(样本阳性率100.00%),显微镜旁台面和培养箱水样的细菌阳性检出率分别为75.00%、12.50%。支原体阳性检出率较高的采样点依次为细胞培养箱水样(100.00%)、房间角落(75.00%)、培养箱外侧扶手(62.50%)和安全柜下方落脚处(62.50%)。人员流动频繁的实验室房间角落、房间地面中央,以及安全柜落脚处的细菌数目均多于人员相对稳定的实验室(P<0.05)。3个时间点(紫外照射30 min、无人员操作运行30 min与人员操作30 min后)生物安全柜内表面的样本未培养出细菌,但部分能检出支原体阳性。结论:人员流动频繁的实验室发生污染的可能性更高,生物安全柜隔绝支原体污染的效果欠佳。加强人员管理、建立清洁制度以及加强生物安全柜维护方可有效避免微生物污染。Objective:To provide a reference basis for promoting the standardized management of the laboratory by the understanding of the operation and management of the cell culture laboratory of a university in Guangxi,and the detection of bacteria and mycoplasma pollution.Methods:Eight cell culture laboratories were randomly selected from a university in Guangxi.The total number of bacteria was detected by AGAR bacterial culture method,and mycoplasma contamination was detected by PCR amplification and agarose gel electrophoresis.Results:All the sampling sites investigated in this study had varying degrees of bacterial and mycoplasma contamination,in which bacteria(the sample positive rate was 100.00%)could be detected in the corner of the room,the center of the floor,the foot of the safety cabinet,the outer handrail of the incubator and the inner wall of the incubator.The positive rates of bacteria on the table next to the microscope and in the water samples of incubator were 75.00%and 12.50%,respectively.The sampling sites with higher positive rate of mycoplasma were water sample of cell incubator(100.00%),floor of room(75.00%),outer handrail of incubator(62.50%)and foot of safety cabinet(62.50%).The number of bacteria in the corner of the laboratory room,the center of the floor of the room and the place where the safety cabinet was located in the laboratory with frequent staff mobility was more than that in the laboratory with relatively stable staff mobility(P<0.05).At three time points(after ultraviolet irradiation for 30 minutes,unmanned operation for 30 minutes and human operation for 30 minutes),no bacteria were cultured on the inner surface of the biosafety cabinet,but some of the samples could be positive for mycoplasma.Conclusion:Laboratories with frequent staff mobility are more likely to be contaminated,and biosafety cabinets are not effective in isolating mycoplasma contamination.Strengthening personnel management,establishing cleaning system and strengthening biosafety cabinet maintenance can effectively av

关 键 词：细胞培养 细菌 支原体 生物安全柜 

分 类 号：G647[文化科学—高等教育学] R-4[文化科学—教育学]