流式细胞术快速检测产超广谱β-内酰胺酶细菌的研究  

Rapid detection of extended-spectrum β-lactamase producing bateriae with flow cytometry

作　　者：付亮[1] 龙军[1] 袁小澎[1] 

机构地区：[1]南方医科大学珠江医院医学检验中心,广州510282

出　　处：《广东医学》2011年第4期416-418,共3页Guangdong Medical Journal

基　　金：广东省自然科学基金资助项目(编号:5300469)

摘　　要：目的建立一种基于流式细胞术快速检测细菌是否具有产超广谱β-内酰胺酶(ESBL)的方法。方法对36株临床分离已经确定为产ESBL的菌株,应用流式细胞药敏试验(FCST),分别检测每株细菌头孢他啶、头孢他啶/棒酸作用管的碘化丙啶荧光阳性百分率(PI%),依据统计学方法建立FCM检测产超广谱β-内酰胺酶细菌阴阳性的判断标准。对49株临床分离的大肠埃希菌和肺炎克雷伯菌同时应用NCCLS推荐的确证试验以及建立的流式细胞术检测标准进行双盲测定,采用统计学评价新的检测方法。结果 36株临床已经鉴定为产ESBL菌经两种药物作用后,PI%(CAZ+C)-PI%(CAZ)值各有不同。经统计学分析确定阳性株鉴定标准为PI%(CAZ+C)-PI%(CAZ)≥3.0。建立检测标准后49株临床菌株经双盲测定后对两种方法学进行统计学比较分析,证实两种方法相比灵敏度为78.3%,特异度为80.8%,粗一致性为79.6%,两种方法的检测结果差异无统计学意义。结论流式细胞术检测产超广谱β-内酰胺酶细菌的方法与传统方法检测结果具有一致性,但是更为快速、客观、便于自动化,在联合药敏试验方面有较大的临床应用前景,为流式细胞仪应用到细菌药敏试验奠定基础。Objective Establish a rapid detection method of extended - spectrum β - lactamase （ESBL） producing bateriae with Flow cytometry. Methods Propidium iodide fluorescent positive percentage （ PI% ） of negative control and reaction tubes for governing ceftazidime, ceftazidime/clavulanic acid was detected, respectively, by flow cytometry susceptibility testing （FCST） for definition of standard point of extended - spectrum β - lactamase producing bacteriae using statistical analysis. According to the standard, 9 strains clinical isolated Escherichia coli and Klebsiella pneumoniae were both detected with confirmatory test of the NCCLS and FCST. Results Positive ESBL was defined as the difference of PI% （CAZ ＋ C） and Pl% （CAZ） more than 3.0, with the sensitivity, specificity and consistency were 78. 3, 80. 8 and 79. 6%, respectively, adjusted with conventional method as gold standard. No significant difference was revealed between the two methods in ESBL detection. Conclusion The new method based on Flow cytometry is a rapid, accurate and convenient method in ESBL detection.

关 键 词：流式细胞术 超广谱Β-内酰胺酶 快速检测 联合药敏试验 

分 类 号：R446.5[医药卫生—诊断学]