沙门菌基质辅助激光解吸电离飞行时间质谱方法研究

Detecting Salmonella with matrix assisted laser desorption ionization time of flight mass spectrometry

目的优化基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)法对沙门菌鉴定的条件,研究不同前处理方法对MALDI靶板污染情况,建立安全高效的检测沙门菌的方法。方法应用MALDI-TOF-MS法对4株沙门菌标准菌株进行不同培养条件、样品前处理方式检验,确立优化的鉴定程序,对不同处理方式下MALDI靶板进行检测,采用优化的前处理方法对33株腹泻病人粪便标本中分离的沙门菌进行数据采集和图谱比对,所得鉴定结果与常规血清学分型结果比较。结果不同前处理方法、不同的培养基上沙门菌标准菌株均准确鉴定为沙门菌。33株沙门菌用MALDI-TOF-MS方法全部准确鉴定为沙门菌属,19株沙门菌在种的鉴定水平上与血清分型结果完全一致,14株沙门菌在种的鉴定水平上与血清分型结果不一致。不同前处理方法的MALDI靶板上均未发现细菌生长。结论 MALDI-TOF-MS方法能快速、简便、安全、准确鉴定属水平沙门菌,是一种有效的沙门菌检测鉴定的手段。

In order to optimize the matrix-assisted laser desorption ionization time of flight mass spectrometry（MALDITOF-MS）method for Salmonellaidentification,study the target board pollution situation of different pretreatment methods,and establish a safe and effective method for detection of Salmonella,we applied MALDI-TOF-MS technique to detect 4standard strains of Salmonella under different culture conditions and sample preparation method,and established optimization program.MALDI target board was detected under different preparation treatments.The optimization method was used to detect33 strains Salmonella which isolated from diarrhea patients＇ stool.Identification results were compared with serological results.The study found that MALDI-TOF-MS method could accurate identify of Salmonellain different pretreatment methods and culture medium.Thirty-three strains of Salmonellaidentified by MALDI-TOF-MS method were all accurate appraisal in genus level,19 strains of them were completely consistent with the serotyping identification results,14 strains of them were not consistent with the serotyping identification results.There was no bacteriagrowth on the target board in different pretreatment methods.MALDI-TOF-MS method can in a fast,convenient,safe and accurate way identify Salmonella,and it can become an effective means for identification of Salmonella.