摘要
目的探讨培养基成分、培养时间对基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析真菌的影响以及该法鉴别产毒和非产毒真菌的可能性。方法(1)实验模式菌株红曲菌分别接种于查氏酵母提取粉琼脂培养基(CYA)和麦芽提取粉琼脂培养基(MEA),曲霉属分别接种于CYA、查氏琼脂培养基(CA)、马铃薯-葡萄糖-琼脂培养基(PDA)和麦芽汁琼脂培养基(MA),青霉属分别接种于CA、CYA和MA培养基,镰孢菌属分别接种于CA、MA和PDA培养基,培养10d(红曲菌)和7d后进行MALDI-TOF-MS分析;(2)模式菌株寄生曲霉接种于PDA培养基,在培养5、7、10和14d时进行MALDI-TOF-MS分析;(3)产毒和非产毒(或低产毒)红曲菌和黄曲霉分别接种于MEA和PDA培养基,培养7d后进行MALDI-TOF-MS分析。结果CYA、PDA、CA和MA分别是红曲菌、曲霉属黄绿组和镰孢菌属、曲霉属黑色组、青霉属进行MALDI-TOF-MS分析的最适培养基;真菌进行MALDI-TOF-MS分析的最佳培养时间为7d;从MALDI-TOF-MS质谱图上尚不能区分红曲菌高产毒株和低产毒株;黄曲霉产毒株和非产毒株的质谱图各异。结论MALDI-TOF-MS进行真菌鉴定时必须对培养基、培养时间进行标化。
Objective To study the effect of medium composition and cultivation time on identification of fungi by matrixassisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. To explore the possibility of differentiating toxigenic fungus from nontoxigenic one by MALDI-TOF-MS. Methods （1） Monascus rubber was inoculated on czapek yeast agar （CYA） and malt extract agar （MEA） media, Aspergillus species on czapek agar （CA）, potato dextrose agar （PDA）, malt agar （MA） and CYA media, PeniciUium species on CA, CYA and MA media, Fusar/um species on CA, MA and PDA media, respectively. M. rubber was incubated for 10 days and others for 7 days, respectively and analyzed by MALDI-TOF-MS. （2） A. parasiticus was inoculated on PDA media, incubated for 14 days and analyzed by MALDI-TOF-MS at day 5, day 7, day 10 and day 14, respectively. （3） Toxigenic M. rubber and A. flavus with high and low levels of toxin production as well as nontoxigenic one were inoculated on MEA and PDA media, respectively, incubated for 7 days and analyzed by MALDI-TOF-MS. Results CYA, PDA, CA as well as MA are the optimum media for identification of Moanscus, both Aspergillus olivine group and Fusaruim species, Aspergillus black group and Penicillium species, respectively by MALDI-TOF-MS. The optimum cultivation time for fungi analysis by MALDI-TOF-MS is day 7. It is not easy to differentiate the toxigenic Monascus strains with high levels of citrinin production from those with low levels of citrinin production. But it is quite different in MALDI-TOF-MS spectrum between toxigenic and nontoxigenic A. flavus. Conclusion It is essential to standardize the media and cultivation time in fungal identification by MALDI-TOF-MS.
出处
《中国食品卫生杂志》
2007年第6期481-488,共8页
Chinese Journal of Food Hygiene
基金
国家自然基金资助课题(30471456)
国家"十一五"科技支撑计划资助课题(2006BAK02A27)
关键词
真菌
光谱法
质量
基质辅助激光解吸电离
生物学鉴定法
因素分析
统计学
Fungi
Spectrometry, Mass, Matrix-Assisted Laser Desoption-Ionization
Biological Assay
Factor Analysis, Statistical
