MALDI-TOF MS技术和AFLP技术在肺炎链球菌同源性分析中的应用  

Application of MALDI-TOF MS and AFLP in homology analysis of Streptococcus pneumoniae

作　　者：黄阿环 陈文标 陈清清[4] 雷莹 李湘琪 陈慧玲 HUANG A-huan;CHEN Wen-biao;CHEN Qing-qing;LEI Ying;LI Xiang-qi;CHEN Hui-ling(Department of Inspection and Prevention,Quanzhou Medical College,Fujian362011,China)

机构地区：[1]泉州医学高等专科学校检验预防学院,福建362011 [2]泉州医学高等专科学校高品医学检验研发孵化基地 [3]泉州医学高等专科学校基础医学部 [4]泉州市第一医院 [5]泉州市儿童妇幼保健院

出　　处：《医学动物防制》2022年第3期265-270,共6页Journal of Medical Pest Control

基　　金：福建省中青年教师教育科研项目(JAT160851);泉州市科技计划项目(2016Z057)。

摘　　要：目的探讨基质辅助激光解吸飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF MS)技术和扩增片段长度多态性(amplified fragment length polymorphism, AFLP)技术在肺炎链球菌的鉴定和同源性分析中的应用。方法收集2016年11月—2017年12月泉州市儿童妇幼保健院临床分离的52株肺炎链球菌标本,用全自动细菌鉴定仪和Optochin试验进行鉴定验证,分别使用MALDI-TOF MS技术和AFLP技术进行同源性分析,形成聚类树,研究两种方法在肺炎链球菌同源性分析中的应用。结果 MALDI-TOF MS技术检测52株细菌均为肺炎链球菌。经过IVD MALDI Biotyper软件获得聚类树图,分为两大簇包括Ⅰ型13株、Ⅱ型39株,其中Ⅱ型又分为两小簇(Ⅱa型24株、Ⅱb型15株)。AFLP技术检测52株肺炎链球菌基因组DNA样本,以电泳条带位置计算出样本间的遗传距离,建构样本的系统发育树,分为两大簇包括Ⅰ型1株、Ⅱ型51株,其中Ⅱ型又分为两小簇(Ⅱa型48株、Ⅱb型3株)。结论 MALDI-TOF MS技术能准确、快速鉴定肺炎链球菌;Biotyper软件进行同源性分析十分快速、便捷,按照0.5差异水平,与AFLP技术的结果有一致性,但也存在差异。Objective To investigate the application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS) and amplified fragment length polymorphism(AFLP) in the identification and homology analysis of Streptococcus pneumoniae.Methods A total of 52 strains of Streptococcus pneumoniae isolated from Children’s Maternal and Child Health Hospital in Quanzhou City from November 2016 to December 2017 were collected and identified using automatic bacterial identification device and Optochin test, and homology analysis was performed using MALDI-TOF MS technology and AFLP technology, respectively, to form a clustering tree and study the application of these two methods in the homology analysis of Streptococcus pneumoniae.Results The 52 strains of bacteria detected by MALDI-TOF MS were all Streptococcus pneumoniae.The clustering tree was obtained by IVD MALDI Biotyper software.The clustering tree was divided into two large clusters(13 of type I and 39 of type II),and two small clusters(24 of type II and 15 of type II).AFLP technique was used to examine 52 genomic DNA samples from Streptococcus pneumoniae,and the genetic distance between samples was calculated based on position of electrophoresis bands.The phylogenetic tree of samples was constructed and divided into two clusters(1 strain of type Ⅰ and 51 strains of type Ⅱ),type II was further divided into two small clusters(48 of type Ⅱa and 3 of type Ⅱb).Conclusion MALDI-TOF MS technology can identify Streptococcus pneumoniae accurately and rapidly;the homology analysis performed by Biotyper software was very fast and convenient.According to the difference level of 0.5,the results were consistent with those of AFLP technology, but there were also differences.

关 键 词：基质辅助激光解吸飞行时间质谱 扩增片段长度多态性 肺炎链球菌 同源性 应用 分析 

分 类 号：R378.12[医药卫生—病原生物学] R446.5[医药卫生—基础医学]