建立一种适合本实验室快速检测产KPC酶肺炎克雷伯菌的方法  

Establishment of a suitable method for rapid detection of KPC producing Klebsiella pneumoniae in our laboratory

作　　者：滕元姬[1] 易雪丽[1] 陈晓颖 罗斌[1] 王春芳[1] TENG Yuanji;YI Xueli;CHEN Xiaoying;LUO Bin;WANG Chunfang(Department of Clinical Laboratory,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院附属医院检验科,广西百色533000

出　　处：《右江医学》2024年第8期740-745,共6页Chinese Youjiang Medical Journal

基　　金：2024年度广西高校中青年教师科研基础能力提升项目(2024KY0545);右江民族医学院附属医院2022年中青年骨干人才科研项目(Y202210302);2021年度百色市科学研究与技术开发计划课题(百科字〔2021〕18号);广西壮族自治区卫生和计划生育委员会自筹经费科研课题(Z20170232)。

摘　　要：目的运用基质辅助激光解吸电离飞行时间质谱技术(MALDI-TOF MS)检测产肺炎克雷伯菌碳青霉烯酶(KPC)耐碳青霉烯类肺炎克雷伯菌(CRKP),筛选并验证其特征峰,建立一种适用于本实验室快速检测产KPC酶型CRKP的方法。方法收集临床耐碳青霉烯类肺炎克雷伯菌39株,运用PCR方法检测出其中23株产KPC酶,再采用MALDI-TOF MS技术检测KPC酶阳性CRKP菌株,得到质谱图后用Flex Analyst软件分析筛选出其中特征峰,并进行重复性实验和临床验证评估。结果综合筛选出产KPC酶CRKP最佳特征峰为m/z 6432,特征峰验证实验和重复性实验的特异性和敏感性分别为96.67%和100%。临床验证评估特异性和敏感性分别为92.31%和100%。结论利用MALDI-TOF MS技术可检出产KPC酶CRKP的最佳特征峰为m/z 6432,经实验证实该特征峰特异性和敏感性高,可为临床快速诊断和治疗产KPC酶CRKP提供理论基础。Objective To detect Klebsiella pneumoniae carbapenemase(KPC)producing carbapenem-resistant Klebsiella pneumoniae(CRKP)by matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),and to screen and verify its characteristic peak,so as to establish a suitable method for rapid detection of KPC-producing CRKP in our laboratory.Methods 39 clinical strains of CRKP were collected,and 23 of them were detected to produce KPC by PCR.MALDI-TOF MS technology was used to detect KPC-positive CRKP strains,Flex Analyst software was used to analyze and screen out their characteristic peaks after the mass spectrum was obtained,and then repeatable experiments and clinical verification evaluations were carried out.Results The best characteristic peak of KPC-producing CRKP was m/z 6432.The specificity and sensitivity of the validation experiment and the repeatability experiment of characteristic peak were 96.67%and 100%,respectively.The specificity and sensitivity of clinical validation evaluation were 92.31%and 100%,respectively.Conclusion The best characteristic peak of KPC-producing CRKP that can be detected by using MALDI-TOF MS technology is m/z 6432.It is proved by experiments that this characteristic peak has high specificity and sensitivity,which can provide a theoretical basis for rapid clinical diagnosis and treatment of KPC-producing CRKP.

关 键 词：基质辅助激光解吸电离飞行时间质谱技术 特征峰 肺炎克雷伯菌 碳青霉烯酶 

分 类 号：R446.1[医药卫生—诊断学]