核酸检测自动化工作站的污染风险及检测性能评估  

Performance and pollution risk assessment of nucleic acid testing automation workstation

作　　者：罗明[1] 王雪 罗琴 李茂中[1] 黄芳[1] 张合润[1] 李爱华[1] 黄琪 龚成[1] LUO Ming;WANG Xue;LUO Qin;LI Mao-zhong;HUANG Fang;ZHANG He-run;LI Ai-hua;HUANG Qi;GONG Cheng(Beijing Cente for Disease Control and Prevention,Beijing Cente for Disease Preventive Medical Research,Beijing 100013,China)

机构地区：[1]北京市疾病预防控制中心/北京市预防医学研究中心,北京100013 [2]首都医科大学公共卫生学院

出　　处：《现代预防医学》2021年第15期2802-2807,共6页Modern Preventive Medicine

基　　金：国家科技部重大专项(2017ZX10103004-002);首都卫生发展科研专项(2020-4-3014)。

摘　　要：目的评价核酸检测自动化工作站的PCR污染风险和检测性能。方法为自动化工作站加配带HEPA滤器的百级层流罩,在仪器舱内形成正压。在层流罩开启和关闭的情况下,通过交叉原则布板评价标本间交叉污染风险,并采集开放水样、环境涂抹样本进行核酸检测,以评估环境污染风险;将脊髓灰质炎病毒Ⅰ型Sabin株系列稀释作为样品,重复测量2次评价其重复性;并采用自动化工作站和现有方法两种方法平行检测上述样品,比较两种方法的检测限;选取183份急性呼吸道患者标本,利用自动化工作站与现有方法同时进行检测,计算检测一致性。结果核酸检测自动化工作站开启层流罩时,216份阴性对照孔与24份环境样本均为阴性,未发现标本间交叉污染及环境污染,关闭层流罩时,216份阴性对照孔共检出11份肠道病毒弱阳性结果,污染率达5.09%(11/216);自动化工作站进行病毒检测的Ct值组间、组内变异系数均小于5%,重复性良好;自动化工作站与现有方法的灵敏度基本相当;利用自动化工作站与现有方法平行检测183份呼吸道标本的总一致性为95.08%,阳性一致性为88.89%,阴性一致性为97.67%。结论自动化核酸检测工作站配置HEPA滤器层流罩,可有效降低PCR污染风险;工作站法的重复性较好、检测限不劣于现有方法,具有较好的推广应用价值。Objective To explore the risk of PCR contamination of the nucleic acid testing automation workstation and its performance.Methods The automation workstation was equipped with a laminar flow hood with HEPA to maintain positive air pressure in the cabin.To assess the risk of cross nucleic-acid contamination between samples by cross-setting the dilutions and water samples at the 96-well lysis buffer plate in the case of laminar flow hood opening and closing.To evaluate the risk of environmental nucleic-acid pollution,four water samples were set at the four corners in the cabin of the workstation and smear samples on the platform outside of the cabin were also collected after the test.To explore the reproducibility of the automation workstation,the concentrated culture of poliovirus SabinⅠstrain was diluted serially for enterovirus testing which was done twice.These dilutions were assayed to compare the lower detection limits by the automation workstation and existing method in parallel.A total of 183 specimens from patients with acute respiratory tract infection were processed and assayed for 14 respiratory viruses by the two mentioned methods to calculate their percentages of agreement.Results 216 negative controls and 24 environmental samples were negative,and no cross-contamination and environmental contamination were found when the laminar flow hood was opened.11 samples were weakly positive for enterovirus in 216 negative control Wells,with a contamination rate of 5.09%(11/216)when the laminar flow hood was closed.The automation workstation showed the excellent reproducibility with<5%of inter-group and intra-group coefficient of variation on Ct value.The sensitivity of automation workstation was almost the same as that of existing method.When 183 respiratory samples being tested using the two mentioned methods,their overall percentage of agreement was 95.08%,their positive percentage of agreement was 88.89%,and the negative percentage of agreement was 97.67%.Conclusion The automation workstation equipped with

关 键 词：PCR实验室自动化工作站 核酸检测 PCR污染 

分 类 号：R115[医药卫生—公共卫生与预防医学]