MALDI-TOF-MS检测哈维弧菌前处理条件的探索及聚类分析  

Exploration of pretreatment conditions and cluster analysis for detection of vibrio harveyi by MALDI-TOF-MS

作　　者：武静[1] 陈英剑[1] 李继霞 胡成进[1] WU Jing;CHEN Yingjian;LI Jixia;HU Chengjin(Departments of Laboratory Medicine,960 Hospital of PLA,Jinan,Shandong 250031,China)

机构地区：[1]中国人民解放军联勤保障部队第九六○医院实验诊断科

出　　处：《国际检验医学杂志》2020年第1期14-17,共4页International Journal of Laboratory Medicine

基　　金：全军医学科技“十二五”重点项目（BWS12J014）

摘　　要：目的 对基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)检测哈维弧菌的前处理条件进行优化,并通过聚类分析初步探讨质谱对哈维弧菌溯源性的价值。方法 本研究首先利用rpoB测序对33株哈维弧菌进行确证,然后利用MALDI-TOF-MS采集哈维弧菌蛋白指纹图谱。并通过培养基的选择、培养时间及样本处理方法对检验前操作程序进行优化。最后,利用MALDI-TOF-MS对33株哈维弧菌进行聚类分析,探讨质谱对细菌溯源性的价值。结果 本研究所用3种培养基的选择对哈维弧菌的鉴定结果差异无统计学意义(P>0.05)。培养时间18、24h哈维弧菌的鉴定结果优于48h的鉴定结果,差异有统计学意义(P<0.05)。但培养18、24h之间鉴定结果差异无统计学意义(P>0.05)。甲酸提取法的效果明显优于直接涂抹法,差异有统计学意义(P<0.05)。基于MALDI-TOF-MS法的聚类分析,能哈维弧菌病感染病原体的溯源提供理论依据。结论 MALDI-TOF-MS法操作简单、快速,可实现对哈维弧菌的准确鉴定和溯源性分析,在海洋渔业和海水养殖业将会发挥重要作用。Objective The pretreatment conditions for detection of Vibrio harveyi by Matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS)were optimized,and the value of mass spectrometry in traceability of Vibrio harveyi was preliminarily discussed by cluster analysis.Methods In this study,33 strains of Vibrio harveyi were first confirmed by rpoB sequencing,and then the protein fingerprints were collected by MALDI-TOF-MS.The operation procedure was optimized through the selection of culture medium,culture time and sample processing method.Finally,33 strains of Vibrio harveyi were clustered by MALDI-TOF-MS to explore the value of mass spectrometry in traceability of bacteria.Results There was no significant difference in the identification result of Vibrio harveyi among the three media(P>0.05).The identification results of Vibrio harveyi at 18 and 24 h were better than that at 48 h(P<0.05).However,there was no significant difference between 18 and 24 h(P>0.05).Formic acid extraction method was better than direct smear method(P<0.05).Cluster analysis based on MALDI-TOF-MS provides a theoretical basis for tracing the pathogens of Vibrio harveyi infection.Conclusion MALDI-TOF-MS method is simple and fast,and can accurately identify Vibrio harveyi and analytical traceability.It will play an important role in marine fisheries and mariculture industry.

关 键 词：基质辅助激光解吸电离飞行时间质谱 哈维弧菌 快速检测 rpoB分析 聚类分析 

分 类 号：R446.5[医药卫生—诊断学]