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目的建立快速检测和鉴定溶藻弧菌的方法,采用基质辅助激光解吸电离飞行时间质谱(MALDITOF-MS)和SARAMIS分析软件,对来自水产品中的溶藻弧菌进行了检测和鉴定。方法将从样品中分离得到的可疑菌落于37℃纯化培养24、48、72 h后,分别用微生物API生化鉴定系统和MALDI-TOF-MS方法进行分析鉴定,并对MALDI-TOF-MS鉴定方法的稳定性、准确性和重复性进行初步探讨。结果不同的培养时间,24、48和72 h对MALDI-TOF-MS检测的蛋白质谱图影响较小,鉴定值分别为94.10%,93.90%,92.70%;同时用微生物API-20E生化鉴定试剂盒进行辅助分析鉴定,培养24 h后,鉴定值为97.7%,T值为0.47,但培养72 h后,鉴定值为52.4%,T值为0.15。同一样品点样2次,得到的6张图谱显示出峰位置基本一致。用标准菌株大肠埃希菌ATCC 8739和溶藻弧菌ATCC 17749纯化培养24 h后,鉴定值分别为99.90%和97.60%。结论与传统的生化鉴定方法相比,MALDI-TOF-MS和SARAMIS软件分析方法具有较高的稳定性、重复性和准确性,可以快速、准确地鉴定溶藻弧菌,可用于水产品中溶藻弧菌的高通量、低成本的快速检测鉴定。
Objective To establish a rapid method for the detection and identification of Vibrio alginolyticus. Vibrio alginolyticus from aquatic products was detected and identified by using matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDITOF-MS) and SARAMIS analysis software. Methods The suspicious colonies isolated from the samples were purified and cultured at 37 ℃ for 24, 48 and 72 h, respectively. The biochemical identification system of microorganism API and MALDI-TOF-MS were used for identification and identification. The MALDI-TOF- The stability, accuracy and repeatability of a preliminary study. Results At different culture time, the protein profiles detected by MALDI-TOF-MS at 24, 48 and 72 h had little effect, with the identified values of 94.10%, 93.90% and 92.70% respectively. At the same time, the microbiological API-20E biochemical identification kit Auxiliary analysis identified, cultured for 24 h, the identification value was 97.7%, T value of 0.47, but after 72 h, the identification value of 52.4%, T value of 0.15. The same sample spot 2 times, the resulting six maps show the peak position is basically the same. After 24 h of purification with standard strains of Escherichia coli ATCC 8739 and Vibrio alginolyticus ATCC 17749, the identified values were 99.90% and 97.60%, respectively. Conclusion Compared with the traditional biochemical identification methods, the MALDI-TOF-MS and SARAMIS software analysis methods have high stability, repeatability and accuracy, which can rapidly and accurately identify Vibrio alginolyticus and can be used in aquatic products High-throughput, low-cost, rapid detection and identification of Alginolyticus.