拉曼镊子结合多元统计方法分析两种人体滴虫的差异性  

Investigation of Biochemical Diversity of Two Species of Trichomonads Based on Raman Tweezers Combined with Multistatistical Analysis

作　　者：黄庶识[1,2] 赖钧灼[1] 梁裕芬[3] 韦俊彬[3] 

机构地区：[1]广西科学院生物物理实验室,南宁530003 [2]昆明理工大学生物工程技术研究中心,昆明650224 [3]广西中医学院基础医学院,南宁530001

出　　处：《分析化学》2011年第4期521-527,共7页Chinese Journal of Analytical Chemistry

基　　金：广西自然科学基金项目(No.0728177);广西中医学院自然科学研究基金(No.P2008099)资助

摘　　要：应用单细胞激光拉曼光谱分析技术,对不同来源的阴道毛滴虫和口腔毛滴虫的拉曼光谱数据进行减背景、平滑、基线校正、归一化等处理后做主成分分析(PCA)和辨别函数分析(DFA)等多元统计分析。平均光谱和PCA分析结果表明,阴道毛滴虫和口腔毛滴虫差异最为明显是1002 cm-1峰,其次,差异相关性最大的还有9个拉曼谱峰785,888,979,1127,1287,1317,1364,1446和1465 cm-1,其中4个峰来自核酸,6个峰归属于蛋白质信号峰,反映两种滴虫的蛋白质、核酸的相对含量差异较大,可以通过PCA方法鉴别两种毛滴虫的差异。由于口腔毛滴虫或阴道毛滴虫虫株间差异并不明显,应用PCA结合DFA多元统计学方法,在保留原有数据信息基础上,通过扩大组间差异,缩小组内差距,同种毛滴虫虫株间在一定程度得到区分。口腔毛滴虫4个虫株之间有12个峰是差异最大的谱峰,阴道毛滴虫4个虫株之间有14个峰是差异最大,反映了两种毛滴虫虫株之间核酸、蛋白质、脂类及糖类等生物大分子组成相对含量差异。此外,根据DFA中虫株间欧氏距离聚类,可以确知虫株之间的差异。To investigate the biochemical components and structure between two species of trichomonads,a Raman tweezers was used to collect Raman spectra of single Trichomonas tenax and Trichomonas vaginalis cells taken from different patients,in which multiple statistical analysis,principal component analyses（PCA） and discriminant function analysis（DFA）,were applied to distinguish the biological diversity between tow species of trichomonads and among their strains statistically.A laser beam was introduced into a sample pool on the stage of an inverted microscopy to form an optical trap,a uniform trichomonad with vitality was trapped randomly in water and the Raman scatter was collected,subsequently,Raman data were background-subtracted smoothed,baselined,normalized,PCA,DFA and hierarchical cluster analysed successively.The results showed that Raman spectra scattered from the trapping trichomonad cells could provide intrinsic molecular information,T.tenax and T.vaginalis could be discriminated using PCA based on the difference of 10 most contributed bands,1002,785,888,979,1127,1287,1317,1364,1446 and 1465 cm-1,among which,4 bands were from lipoid,6 bands were from protein;There were no significant differences between those strains of T.tenax or T.vaginalis if the data were analyzed with PCA only,however,4 strains of T.tenax or T.vaginalis could be distinguished easily using DFA based on the principal components（PCs） of pre-PCA,which could maximize between group and minimize within group distances;among the four strains of T.tenax,12 Raman bands were contributed to the difference and 14 bands of T.vaginalis strains were difference significantly as well,in which Raman bands correspond to various biomacromolecule including proteins,lipid,nucleic acid and carbohydrates.Moreover,according to the Euclidean distance in hierarchical cluster analysis（HCA） based on DFA,the difference range between the strains can be ascertained.

关 键 词：口腔毛滴虫 阴道毛滴虫 拉曼光谱 多元统计分析 

分 类 号：R382.2[医药卫生—医学寄生虫学]