创伤弧菌脂肪酸色谱鉴定法临床应用.docVIP

脂肪酸气相色谱技术鉴定创伤弧菌 王志刚 邵平扬 徐幼平 摘要 目的:建立细菌脂肪酸气相色谱分析技术对创伤弧菌(VV)鉴定的新方法。方法:创伤弧菌经35oC、24 h血平板培养，收集菌落，皂化、甲基化、萃取洗涤99.17%的峰能明确识别，VV的脂肪酸种类与含量为 C16：0 (29.66%) ,C16:1ω7C/C16:1ω6C(28.32%), C18:1ω7C (13.42%), C14:0 (5.41%), C14:0 3OH/C16:1 iso 1(4.31%) and C12:0 3OH (4.01%)。结论: 细菌脂肪酸的种类与数量可作为细菌鉴定特征，气相色谱分析技术具有灵敏、精确、快速特点，与经典的细菌生化反应及DNA鉴定相比，具有生物安全、成本低廉等优点，为临床实验室细菌的鉴定开辟了新途径。 [关键词]：创伤弧菌，脂肪酸; 气相色谱法 Identification of Vibrio Vulnificus by fatty acid analysis with Gas Chromatography Zhigang Wang, Pinyang Shao，Youping Xu. Department of clinical laboratory, The second hospital of Jiaxing, Zhejian province, Jiaxing,341000，china [Abstract] Objective: To establish new method for identification of Vibrio Vulnificus(VV) by fatty acid analysis with gas chromatography(GC). Methods VV strain were cultivated on sheep blood agar 35oC for 24 hours. By Harvesting, Saponification，methylation，extraction and basic wash steps. The whole cellular fatty acids were detected by gas chromatography. We matched Peak position of sample results with reference chromatograms. Results The rapid analysis extract to identification VV within 20 minutes. Peak position error were 0.21%.Percent named were 99.17%. bacteria extracts contain straight and branched fatty acids with 9 to 20 carbon lengths. Strain VV summed feature contained C16：0(29.66%),C16:1ω7C/C16:1ω6C(28.32%), C18:1ω7C (13.42%), C14:0 (5.41%), C14:0 3OH/C16:1 iso 1(4.31%) and C12:0 3OH (4.01%) as the major fatty acids. Conclusions According to the qualitative and quantitative differences in fatty cell profiles can differentiate species and sub-species, which combines highly sensitivity, accurate, rapid microbial identification, less expensive and more safety than conventional biochemical/ DNA-based identification. . We presented a novel fatty acid analysis methodology for identification VV. [Key words] Vibrio Vulnificus（VV）;Fatty acid（FA）; Gas Chromatography（GC） 2006年8月，美国《Emerging infectious diseases》杂志将创伤弧菌（Vibrio Vulnificus，