基于质谱技术的婴幼儿肠道菌群多样性鉴别方法研究

Analysis of microorganism in infant feces using HPLC-MS and MALDI-TOF MS

利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF)和液相色谱质谱联用技术(HPLC-MS)对婴幼儿粪便中的微生物组成进行了分析。100例3岁以内不同年龄段的婴幼儿粪便经过预处理后利用两种不同的质谱技术进行了检测,并对两种检测方法的结果进行了比较。结果表明,2~3岁年龄段的婴幼儿检出峰个数最多,0~1岁年龄段的婴幼儿粪便样本中检测出的分子量分布范围最广。利用HPLC-MS分别对不同年龄段的样本进行微生物种类识别。结果表明:0~1岁婴幼儿粪便样本中主要有大肠埃希氏菌、链霉菌、酵母菌、白色念珠菌、链球菌等;1~2岁婴幼儿粪便中主要有红色毛麟菌、大肠埃希氏菌、白色念珠菌、酵母菌等;2~3岁婴幼儿粪便中主要有双歧杆菌、链霉菌、假丝酵母、大肠埃希氏菌等。MALDI-TOF具有检测速度快的优点,但对于未培养的微生物方法的重复性和稳定性不高。HPLC-MS准确度高,可检测出的微生物种类明显高于MALDI-TOF,适用于未经培养的微生物体系。

Microorganism in infant feces was analyzed using matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)and high performance liquid chromatography mass spectrometry(HPLC-MS).Seventy samples from 0 to 3 years old infants were pretreated and analyzed by the two methods,and the results were compared.MALDI-TOF mass spectra indicated that ions observed increased with the infants′age,more microorganisms were detected in 2-3 years old infant feces than that from 0-2 years old.HPLC-MS results indicated that the main microorganism in 0-1years old infant feces were Escherichia coli(E.coli),Streptomycetaceae,yeast,monilia albican and streptococcus.The high abundance microorganism in 1-2 years old infant feces were red hair fungus bacteria,E.coli,monilia albican,yeast and and 2-3 years old infant feces were bifidobacterium,streptococcus,candida and E.coli.