霍乱弧菌基质辅助激光解吸飞行时间质谱数据库的建立与应用
目的 建立基质辅助激光解吸电离飞行时间质谱法(matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-TOF-MS)快速检测霍乱弧菌, 并建立数据库。方法 考察培养基、培养时间、培养温度以及样品前处理方法对图谱质量和鉴定结果的影响, 对方法的稳定性、重复性进行研究; 采集101株霍乱弧菌食品分离株MALDI-TOF-MS指纹图谱, 并对图谱进行分析。其中标准菌株CICC23794及40株分离株作为建库菌株, 其余60株用于建库后的测试。结果 采用2%NaCl-TSA培养基, 在37 ℃、24 h条件下培养, 可达到强峰值信号, 谱图重现性良好, 甲酸-乙腈提取法获得的质谱图特征峰多, 信噪比高; 建库后, 60株霍乱弧菌均可达到高于2.0的水平, 其中约有20株霍乱弧菌鉴定分值可达到2.4以上。结论 本研究建立的霍乱弧菌MALDI-TOF-MS鉴定方法结果准确, 可用于实际检测。
Objective To establish a rapid method for the detection and identification of Vibrio cholerae by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and establish the database. Methods In order to verify the stability and repeatability, different mediums, culture times and sample handing methods were optimized. The 101 isolates of Vibrio cholerae was used as a reference to evaluate the credibility of MALDI-TOF-MS method and analysis on mass spectrometric profiles. The reference strain CICC23794 and 40 isolates were used as the library-building strains, and the other 60 strains were used for the test after the library-building. Results Taking 2%NaCl-Tryptic Soy Agar Base as the culture medium, the mass spectrum with strong signal and good repeatability was acquired after 37 ℃, 24 h. The results showed that more characteristic peak of protein with smooth baseline, less noise and higher signal-to-noise ratio were obtained by ethanol/ formic acid treatment method. The 60 isolates of Vibrio cholera could be identified with a score of 2.0, and 20 strains of Vibrio cholerae could be identified with a score of 2.4 or higher. Conclusion MALDI-TOF-MS is a fast and accurate method for the identification of Vibrio cholera, and can be used for routine detection.
标题：霍乱弧菌基质辅助激光解吸飞行时间质谱数据库的建立与应用
英文标题：Establishment and application of matrix-assisted laser desorption ionization-time of flight mass spectrometry database for the detection of Vibrio cholerae
作者：
刘莉 北京海关技术中心
韩笑 北京海关技术中心
王紫薇 北京海关技术中心
汪琦 北京海关技术中心
赵晓娟 北京海关技术中心
曾静 北京海关技术中心
中文关键词:基质辅助激光解吸飞行时间质谱法,霍乱弧菌,鉴定,
英文关键词:matrix-assisted laser desorption ionization time-of-flight mass spectrometry,Vibrio cholerae,identification,
发表日期：2020-05-03
- 文件大小:
- 5.91 MB
- 下载次数:
- 60
-
高速下载
|
|
