基于AIE技术构建快速细菌药物敏感性试验新方法  

Rapid detection of the bacterial drug susceptibility testing based on AIE technology

作　　者：赖丽莎 邓任堂[1] 张露 揭育帮 谢岭平[1] 黄志宏[1] 尹丽明 王杜娟[1] 李丽娟[2] 徐军发 彭兰芬[1] 付文金[1] Lai Lisha;Deng Rentang;Zhang Lu;Jie Yubang;Xie Lingping;Huang Zhihong;Yin Liming;Wang Dujuan;Li Lijuan;Xu Junfa;Peng Lanfen;Fu Wenjin(Department of Laboratory,Dongguan Houjie hospital,Dongguan 523945,China;Department of Laboratory,Dongguan People′s Hospital,Dongguan 523059,China;Institute of Inspection Medicine,Guangdong Medical University,Dongguan 523808,China)

机构地区：[1]广东省东莞市厚街医院检验科,东莞523945 [2]广东省东莞市人民医院检验科,东莞523059 [3]广东医科大学检验医学研究所,东莞523808

出　　处：《中华检验医学杂志》2023年第11期1186-1192,共7页Chinese Journal of Laboratory Medicine

基　　金：广东省基础与应用基础研究基金(2019B1515120004);东莞市社会科技发展局重点项目(202050715023181)。

摘　　要：目的基于AIE荧光探针6PD-DPAN可结合细菌并聚集发光,荧光强度可体现细菌数量的原理,建立一种快速、简便、准确的细菌药物敏感性试验新方法,为临床快速精准用药提供依据。方法评价类研究。收集东莞市厚街医院2022年1—12月临床样本分离菌共107株,其中46株用于新方法的建立,61株用于方法学验证。微量肉汤稀释法测定的最低抑菌浓度(MIC)为金标准,以庆大霉素,左氧氟沙星、头孢噻肟3种抗菌药物作为实验药物;计算第4 h荧光强度比值(R值),采用ROC曲线分析R值在判断细菌生长与否的效能并确定其截断值。新方法检测61株临床菌株的MIC,以微量肉汤稀释法为金标准,分析新方法的基本一致性、分类一致性、极重大错误和重大错误,采用Kappa检验确定两者的一致性。结果培养4 h R值的ROC曲线分析:截断值为3.0,其对细菌生长的判定敏感度和特异度均为100%;细菌生长受抑制R值为11.1(8.6,14.4);细菌生长R值为1.1(1.0,1.2)。与金标准相比,新方法检测61株临床菌株MIC的基本一致性为100%(61/61),分类一致性为96.7%(59/61),未出现极重大错误和重大错误,Kappa值为0.94,新方法与微量肉汤稀释法结果一致性较好。结论本研究成功构建了一种基于AIE技术的细菌药物敏感性试验新方法,可在5 h内取得满意结果,为临床早期精准药物治疗提供依据。Objective Based on the principle that the aggregation-induced emission(AIE)fluorescent probe 6PD-DPAN could bind and aggregate with bacteria,and the fluorescence intensity could reflect the quantity of bacteria,a new method for rapid,convenient,and accurate bacterial drug sensitivity testing was established,which provided a basis for rapid and accurate clinical drug use.Methods This was a methodological evaluation study.A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022,among which 46 isolates were used for the establishment of the new method,and 61 isolates were used for methodological validation.The minimum inhibitory concentration(MIC)determined by broth microdilution method was used as the gold standard,and three antibacterial drugs,gentamicin,levofloxacin,and cefotaxime,were used as experimental drugs.The AIE plate was incubated for 4 hours,and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve.The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive,intermediate,or resistant.To simplify the detection process,the ratio of fluorescence intensity at 4 hours(R)was calculated,and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value.The new method was used to determine the MIC of 61 clinical isolates,with broth microdilution method as the gold standard.The basic consistency,categorical consistency,very major errors,and major errors of the new method were analyzed,and the consistency between the two methods was determined by the Kappa test.Results ROC curve analysis of the R after 4 hours of culture:The cut-off value was 3.0,with both sensitivity and specificity for determining bacterial growth being 100%.The median(interquartile)R for bacterial growth inhibition was 11.1(8.6,14.4);the median R-value for bacterial growth was 1.1(1.0,1.2).Compared to the gold standard,the newly established method showed 100%(6

关 键 词：微生物敏感性试验 聚集诱导发光 细菌感染 

分 类 号：R446.5[医药卫生—诊断学]