摘要
目的用基质辅助激光解吸/电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry,MALDI-TOF MS)仪构建海南病原类鼻疽伯克霍尔德菌鉴定数据库,评价自建库鉴定准确性并与国外同类产品进行比较。方法海南病原类鼻疽伯克霍尔德菌质谱数据库的构建:收集海南患者标本分离10株类鼻疽伯克霍尔德菌用于建库,以VITEK MS RUO模式进行图谱采集及数据库构建,每株菌平行涂布共12个靶点,得到120张质谱图;分别将10株类鼻疽伯克霍尔德菌经35℃和25℃培养24 h,每株菌每个血平板和巧克力平板分别涂3个靶点,共涂了120个靶点,每个靶点采集1次,得到120张质谱图。培养48 h,再次进行同样的靶点采集120张图谱。将采集获得240张图谱按标准精选得到优质图谱109张。通过仪器建库参数确认,控制原始波峰数量,筛选特征峰并赋权予权重,调整质量数、谱峰强度在规定范围,经过SARAMIS软件分析生成海南病原类鼻疽伯克霍尔德菌数据库。数据库验证:收集海南不同地区来源患者类鼻疽伯克霍尔德菌30株及其他伯克霍尔德菌20株,以16SrRAN基因测序为金标准,比较自建数据库与VITEK MS v 4.16数据库鉴定类鼻疽伯克霍尔德菌的准确性及图谱特征。结果构建的海南病原类鼻疽伯克霍尔德菌质谱数据库经30株类鼻疽伯克霍尔德菌验证菌株确认,基于自建数据库,种水平鉴定准确率为100.0%(30/30),高于VITEK MS v 4.16数据库准确率56.67%(17/30),差异有统计学意义(χ^(2)=11.08,P<0.005)。结论构建的海南病原类鼻疽伯克霍尔德菌质谱数据库质量达到临床常规鉴定应用要求,就海南病原流行株的鉴定准确率优于国外同类产品。
Objective To construct the identification database of pathogenic Burkholderia pseudomallei in Hainan with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS),and to evaluate the identification accuracy of the self-built database and compare it with similar products abroad.Methods Hainan pathogenic Burkholderia pseudomallei mass spectrometry database was constructed.Ten strains of Burkholderia pseudomallei isolated from Hainan patient specimens were collected,and the atlas was collected and the database was constructed by VITEK MS RUO mode.Each strain was coated with 12 targets in parallel,and 120 mass spectra were obtained.Ten Burkholderia pseudomallei strains were cultured at 35℃and 25℃for 24 h,respectively;and each blood plate and chocolate plate of each strain were coated with three targets,with a total of 120 targets.Each target was collected once,and 120 mass spectra were obtained.After 48 hours culture,120 maps of the same target were collected again.A total of 240 atlas were collected and selected according to the standard,and so as 109 high-quality atlas were obtained.Through the confirmation of instrument database building parameters,the number of original peaks was controlled,the characteristic peaks were screened and weighted,the mass number and spectral peak intensity were adjusted within the specified range,and the super database of pathogenic Burkholderia pseudomallei in Hainan was generated after analysis by SARAMIS software.Thirty strains of Burkholderia pseudomallei and 20 strains of other Burkholderia from patients in different areas of Hainan were collected,and the accuracy and map characteristics of identification of Burkholderia pseudomallei were compared be-tween self-built database and VITEK MS v4.16 database with 16SrRAN gene sequencing as the gold standard.Results The constructed Hainan pathogenic Burkholderia pseudomallei mass spectrometry database was confirmed by 30 verified strains of Burkholderia pseudomallei.Based on the self-built database,the accuracy of species level identification was 100.0%(30/30),significantly higher than Vitek MS v4.16 database accuracy 56.67%(17/30)(χ^(2)=11.08,P<0.005).Conclusion The quality of the constructed pathogenic Burkholderia pseudomallei mass spectrometry database in Hainan meets the application requirements of clinical routine identification,and the identification accuracy of Hainan epidemic strains is better than that of similar foreign products.
作者
陈如寿
钟佳芳
石挺丽
吴吉芳
王婧婧
颜礼完
CHEN Ru-shou;ZHONG Jia-fang;SHI Ting-li;WU Ji-fang;WANG Jing-jing;YAN Li-wan(Department of Laboratory,Sanya Central Hospital(Hainan Third People's Hospital),Sanya 572000,Hainan,China;不详)
出处
《广东医学》
CAS
2022年第6期707-711,共5页
Guangdong Medical Journal
基金
海南省卫生计生行业科研项目(19A200150,19A200120)。
关键词
类鼻疽伯克霍尔德菌
光谱法
质量
基质辅助激光解吸电离
数据库
鉴定
Burkholderia pseudomallei
spectrometry
mass
matrix-assisted laser desorption away from
database
authenticate
