摘要
目的探索基于环介导等温扩增技术(loop-mediated isothermal amplification, LAMP)的离心式微流控芯片在食源性致病菌快速检测中的应用价值。方法对2022年1月至2024年1月延安市疾病预防控制中心收集的128份检验样本进行回顾性分析, 均进行细菌培养、LAMP的离心式微流控芯片检测, 比较两种检测方式的检测时间、加标浓度以及准确率, 且经受试者工作特征曲线(receiver operating characteristic curve, ROC)分析两种检测方式诊断效能。结果细菌培养大多集中在16~48 h检测出, 检出限均集中于10 000 CFU/25 g, LAMP的离心式微流控芯片大多集中在5~16 h检测出, 检出限集中于1 000~10 000 CFU/25 g, 差异均有统计学意义(t=14.86、573.04, 均P < 0.001)。同时, 与后期确诊结果比较, 细菌培养检出阳性率75.00%(24/32), 诊断准确率80.47%(103/128), 一致性较低(Kappa=0.51, P < 0.001);LAMP的离心式微流控芯片检出阳性率93.75%(30/32), 诊断准确率96.87%(124/128), 一致性较低(Kappa=0.92, P < 0.001)。两两比较中, LAMP的离心式微流控芯片与细菌培养比较, 差异有统计学意义(χ^(2)=53.14, P < 0.001)。而经ROC曲线分析, 细菌培养、LAMP诊断食源性致病菌的AUC分别为0.77、0.95。结论在检测食源性致病菌时, 运用LAMP的离心式微流控芯片技术, 更能够在短时间内检测出致病菌, 具有高效、快速、准确率高等优势。
Objective:To investigate the application value of centrifugal microfluidic chips based on loop-mediated isothermal amplification(LAMP)technology for rapid detection of foodborne pathogens.Methods:A retrospective analysis was conducted on 128 test samples collected by the Yan'an Center for Disease Control and Prevention from January 2022 to January 2024.All samples underwent bacterial culture and testing using centrifugal microfluidic chips based on LAMP technology.The detection time,spiking concentration,and accuracy of two detection methods were compared.Additionally,the diagnostic efficacy of both methods was analyzed using receiver operating characteristic(ROC)curves.Results:Bacterial cultures were mostly detected within 16-48 hours,with a detection limit concentrated at 10000 CFU/25 g.In contrast,the LAMP-based centrifugal microfluidic chip technology typically yielded results within 5-16 hours,with a detection limit ranging from 1000 to 10000 CFU/25 g.The differences were statistically significant(t=14.86,573.04,both P<0.001).When compared with subsequent confirmation results,the positive rate for bacterial culture was 75.00%(24/32),with a diagnostic accuracy of 80.47%(103/128),showing a low consistency(Kappa=0.51,P<0.001).Meanwhile,the LAMP-based centrifugal microfluidic chip technology had a positive detection rate of 93.75%(30/32)and a diagnostic accuracy of 96.87%(124/128),also exhibiting low consistency(Kappa=0.92,P<0.001).In paired comparisons,the difference between the LAMP-based centrifugal microfluidic chips and bacterial culture was statistically significant(χ^(2)=53.14,P<0.001).ROC curve analysis indicated that the area under the curve for bacterial culture in diagnosing foodborne pathogens was 0.77,while the AUC for LAMP-based centrifugal microfluidic chip technology was 0.95.Conclusions:When detecting foodborne pathogens,the use of LAMP-based centrifugal microfluidic chip technology allows for the rapid identification of pathogens in a short time period,offering advantages such as high efficiency,speed,and accuracy.
作者
杨蒙
刘玮
白跳
王玲玲
Yang Meng;Liu Wei;Bai Tiao;Wang Lingling(Department of Laboratory Medicine,Yan'an Center for Disease Control and Prevention,Yan'an 716000,ShaanxiProvince,China)
出处
《中国基层医药》
2025年第2期250-255,共6页
Chinese Journal of Primary Medicine and Pharmacy
关键词
食物传播疾病
细菌
培养技术
ROC曲线
芯片分析技术
Foodborne diseases
Bacteria
Culture techniques
Roc curve
Microchip analytical procedures
