基质辅助激光解吸电离飞行时间质谱法检测副溶血性弧菌

Detection of Vibrio parahaemolyticus by matrix assisted laser desorption ionization time of flight mass spectrometry

中文关键词:  基质辅助激光解吸电离飞行时间质谱法  副溶血性弧菌  快速检测

英文关键词:matrix assisted laser desorption ionization time of flight mass spectrometry  Vibrio parahaemolyticus  rapid detection

中文摘要:

      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)检测副溶血性弧菌的方法。方法 从样品预处理方法、分析菌量、培养基、培养时间方面分析各因素对MALDI-TOF-MS方法重复性、鉴定结果准确性的影响。将从市售420份水产品中分离并用全自动微生物鉴定及药敏分析系统生化鉴定为副溶血性弧菌的123株菌株, 用MALDI-TOF-MS对其进行鉴定, 对比2种方法的鉴定结果。结果 采用混合溶液法进行预处理, 3% NaCl营养琼脂培养基, 36 ℃培养24 h, 分析菌量为8 mg, 用MALDI-TOF-MS进行分析所得到的特征峰多、峰型稳定、杂峰少, 质谱图的准确性和重复性高、与生化鉴定匹配性好。分离得到的123株副溶血性弧菌鉴定分值大于2.000的达98.4%。结论 MALDI-TOF-MS作为一种快速、准确、高效的检测方法, 可用于食品中副溶血性弧菌快速检测。

英文摘要:

      Objective To establish a method for the detection of Vibrio parahaemolyticus by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Methods The effects of various factors such as pretreatment method, analytical bacterial count, medium and culture time on the repeatability and accuracy of MALDI-TOF-MS method were studied. A total of 123 strains isolated from 420 commercial aquatic products and identified as Vibrio parahaemolyticus by automatic microbial identification and drug sensitivity analysis system were identified by MALDI-TOF-MS, and the identification results of the 2 kinds of methods were compared. Results Taking 8 mg bacteria of 3% NaCl nutrient agar, the mass spectrum with stable peak shape and strong signal was acquired after 37 ℃, 24 h, more characteristic peaks and few miscellaneous peaks were obtained by mixed solution method, the mass spectrum has high accuracy and repeatability, and matched well with biochemical identification. The identification value of 123 strains of Vibrio parahaemolyticus isolated was more than 2.000 (98.4%). Conclusion MALDI-TOF-MS is a rapid, accurate and efficient detection method, which can be used for rapid detection of Vibrio parahaemolyticus in food.