傅里叶变换红外光谱技术对4种志贺氏菌快速分型的研究  

Rapid Typing Method for Four Species of Shigella by FT-IR Spectroscopy

作　　者：李兆杰 张玉春 刘玉敏 江艳华[2] 王秀杰 顾华珍 李正义[3] 杨丽君 LI Zhao-Jie;ZHANG Yu-Chun;LIU Yu-Min;JIANG Yan-Hua;WANG Xiu-Jie;GU Hua-Zhen;LI Zheng-Yi;YANG Li-Jun(Weihai Entry-Exit Inspection and Quarantine Bureau,Weihai 264205,China;Yellow Sea Fisheries Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,Chin;Shandong Entry-exit Inspection and Quarantine Bureau,Qingdao 266002,China)

机构地区：[1]威海出入境检验检疫局,山东威海264205 [2]中国水产科学院黄海水产研究所,山东青岛266071 [3]山东出入境检验检疫局,山东青岛266002

出　　处：《中国海洋大学学报（自然科学版）》2018年第1期57-62,共6页Periodical of Ocean University of China

基　　金：国家质量监督检验检疫总局科研项目(2015IK204;2015IK203)资助~~

摘　　要：应用傅立叶变换红外光谱(fourier transform infrared spectroscopy,FT-IR)技术,采集志贺氏菌属痢疾志贺氏菌(Shigella dysenteriae,S.dysenteriae)、福氏志贺氏菌(Shigella flexneri,S.flexneri)、鲍氏志贺氏菌(Shigella boydii,S.boydii)、宋内氏志贺氏菌(Shigella sonnei,S.sonnei)4种志贺氏菌的4 000~600cm^(-1)波数范围的中红外光谱,对光谱进行基线校正、矢量归一化等处理;应用主成分分析(Principal component analysis,PCA)和分级聚类分析(Hierarchical cluster analysis,HCA)分别对4种志贺氏菌1 800~900cm^(-1)波数范围的光谱数据进行聚类分析,通过对污染猪肉样品里分离的可疑S.sonnei、肠炎沙门氏菌(Salmonella enteritidis)和出血性大肠埃希氏菌((Escherichia.Coli O157:H7)进行HCA分析来验证所建立的数据库和分型方法。结果显示:成功建立了4种志贺氏菌的红外光谱数据库,可以用于可疑目标菌的快速分型;PCA和HCA 2种聚类方法均能将4种志贺氏菌进行较好分类,而且应用建立的数据库和HCA聚类方法能准确对可疑目标菌进行快速分型,说明FT-IR技术结合化学计量学运算方法,可以将4种志贺氏菌进行快速分型。该研究为志贺氏菌的快速分型提供了一种快速简便的方法,适应于地方疾控部门和口岸检疫部门开展志贺氏菌监测和流行病学溯源工作。The fingerprint absorption spectra from 4 000~600 cm-1 of four species of Shigella including S.dysenteriae,S.flexneri,S.boydii and S.sonnei were collected by FT-IR spcetroscopy,treated by baseline correction,vector normalization,and so on.The spectra data from 1 800~900 cm-1 were analyzed by two unsupervised methods of principal component analysis(PCA)and hierarchical cluster analysis(HCA)respectively to establish a rapid typing method for four species of Shigella.The spectra data library and the typing method established here were tested by HCA of target S.sonnei,S.enteritidis and E.coli seperated from contaminated pork.It was shown that a standard FT-IR spectral data library including four species of Shigella was established and could be used for the rapid typing of Shigella.Furthermore,the two methods of PCA and HCA could give good seperation of the four bacteria.The PCA and HCA results proved that the region over the range of 1 800~900 cm-1 exhibited the most representative differences,which contains three“fingerprint regions”having unique absorption,called the amide region,the mixed region and the polysaccharide region,respectively.It was also revealed that the clustering pattern obtained from HCA was in agreement with the classification obtained using PCA,indicating that the FT-IR-based methods can deliver consistent classification in spite of the multivatiate statistical analysis used.The spectra data of target S.sonnei from contaminated pork was clusterd successfully in the S.sonnei cluster of the spectra data library,but those of S.enteritidis and E.coli was clustered alonely.These results demonstrate that FT-IR spectroscopy technology combined multivariate statistical analysis is a good typing method for four species of Shigella.This study may provide a rapid,accurate and easy method for typing of Shigella,especially suitable for the basic laboratories of CDC and port quarantine departments to perform suiveillance and epidemiological traceability of Shigella.

关 键 词：傅立叶变换红外光谱 志贺氏菌 分型 主成分分析 分级聚类分析 

分 类 号：Q31[生物学—遗传学]