应用MAIDI-TOF MS 分析黄连素作用前后耐碳青霉烯类肺炎克雷伯菌质谱峰图的改变*

目的应用基质辅助激光解吸电离飞行时间质谱仪(matrix-assisted laser desorption ionization time-of-flight mass spectrometry, MALDI-TOF MS)分析产碳青霉烯酶肺炎克雷伯菌被黄连素作用前后蛋白质谱峰的改变。方法 收集浙江中医药大学附属第一医院产碳青霉烯酶（KPC）肺炎克雷伯菌20例，使用培养法得到黄连素作用后传代菌株，应用MAIDI-TOF MS 分组检测每代菌株，,ClinProTools 3. 0 软件对 MALDI-TOF MS 鉴定细菌所产生的质谱峰图进行分析。结果 质谱将黄连素作用前KPC肺炎克雷伯菌和黄连素作用后第三代肺炎克雷伯菌较清晰地划分成两个区域， ClinProTools 软件的3种算法所得出的结果基本一致，灵敏度均大于 95.0%，其中监督式神经网络算法（ＳＮＮ）灵敏度最高，为100.0%，三种算法特异性均大于85.0%。质荷比为15143.96m/ z、 12265.75m/z、16099.26m/z、15048.83m/z的质谱峰是用于区分作用前后最为主要的特征性峰，在峰强度上存在一定的差异。结论 黄连素作用于碳青霉烯类肺炎克雷伯菌后蛋白质谱峰发生了明显的改变。

Analysis of the change of the mass spectrome try of the carboniamian pneumonitis using MAIDI-TOF MS technology

Purpose To use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze carbapenem Klebsiella pneumoniae before and after the action of berberine Changes in protein peaks.Methods Collect 20 cases of Klebsiella pneumoniae producing carbapenemase (KPC) from the First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, and use the culture method to obtain the subcultured strains after berberine action, and use MAIDI-TOF MS to detect each generation of strains, ClinProTools 3.0 software analyzes the mass spectra generated by MALDI-TOF MS to identify bacteria.Results Mass spectrometry divided KPC pneumoniae before the action of berberine and the third generation of Klebsiella pneumoniae after the action of berberine. The results obtained by the three algorithms of ClinProTools are basically the same, and the sensitivity is basically the same. All are greater than 95.0%, among which the supervised neural network algorithm (SNN) has the highest sensitivity, which is 100%, and the specificity of the three algorithms is greater than 85%. The mass spectrum peaks with mass-to-charge ratios of 15143.96m/z, 12265.75m/z, 16099.26m/z, 15048.83m/z are the most important characteristic peaks used to distinguish before and after the action, and there is a certain difference in peak intensity.Conclusion After berberine acted on carbapenem-like Klebsiella pneumoniae, the protein peak of the protein spectrum changed significantly.