Confocal Raman microspectroscopy combined with chemometrics as a discrimination method of clostridia and serotypes of Clostridium botulinum strains,Journal of Raman Spectroscopy

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Confocal Raman microspectroscopy combined with chemometrics as a discrimination method of clostridia and serotypes of Clostridium botulinum strains
Journal of Raman Spectroscopy ( IF 2.4 ) Pub Date : 2021-09-08 , DOI: 10.1002/jrs.6244
Jin Zhang _{1,

2} , Hong Jiang ₁ , Pengya Gao ₂ , Yuan Wu ₂ , Hui Sun ₂ , Ying Huang ₂ , Xuefang Xu ₂

Affiliation

Rapid and accurate identification of Clostridium botulinum is of great importance because it has been considered as an emerging food-borne pathogen and potential zoonotic agent. Raman spectroscopy can differentiate bacteria based on Raman scattering spectral patterns of whole cells in a fast, reagentless, and easy-to-use manner. This study demonstrates that confocal Raman microspectroscopy (CRM) combined with chemometrics can serve as a fast, reliable, and nondestructive method for detection and identification of C. botulinum at both species and serotypes level without any laborious pre-treatments. Three significant bacillus pathogens including C. botulinum, C. perfringens, and C. difficile were investigated with CRM. Additionally, two main C. botulinum strains causing botulism, C. botulinum type A, and C. botulinum type B were examined with CRM. Principal component analysis (PCA) was performed to differentiate the three species. PCA and linear discrimination analysis (LDA) were used for serotyping C. botulism strains. Four common and important preprocessing methods including Savitzky–Golay algorithm smoothing (SG), standard normal variate (SNV), multivariate scatter correction (MSC), and Savitzky–Golay algorithm 1st Derivative (SG 1st Der) were applied to improve the accuracy of identification and explore the impact of various single preprocessing methods on the model. The results proved that CRM coupled with chemometrics can be utilized for fast, reliable, and nondestructive identification of clostridia and serotypes of C. botulinum strains. This study proves for the first time that the CRM combined with chemometrics methods can be used as a potential means to detect and identify C. botulinum.

中文翻译：

共聚焦拉曼显微光谱结合化学计量学作为梭菌和肉毒梭菌菌株血清型的鉴别方法

快速准确地鉴定肉毒杆菌非常重要，因为它被认为是一种新兴的食源性病原体和潜在的人畜共患病原体。拉曼光谱可以快速、无试剂且易于使用的方式基于全细胞的拉曼散射光谱模式区分细菌。本研究表明，共聚焦拉曼显微光谱 (CRM) 结合化学计量学可以作为一种快速、可靠和无损的方法，用于在物种和血清型水平上检测和鉴定肉毒杆菌，无需任何繁琐的预处理。肉毒梭菌、产气荚膜梭菌和艰难梭菌等三种重要的芽孢杆菌病原体用CRM进行了调查。此外，两个主要的肉毒梭菌造成肉毒杆菌菌株，肉毒梭菌键入甲，和肉毒梭菌型乙用CRM检查。进行主成分分析 (PCA) 以区分这三个物种。PCA 和线性判别分析 (LDA) 用于血清分型肉毒杆菌菌株。应用Savitzky-Golay算法平滑(SG)、标准正态变量(SNV)、多元散射校正(MSC)、Savitzky-Golay算法一阶导数(SG 1st Der)四种常用且重要的预处理方法提高识别精度并探索各种单一的预处理方法对模型的影响。结果证明，CRM 结合化学计量学可用于快速、可靠和无损地鉴定梭菌和肉毒梭菌菌株的血清型。本研究首次证明CRM结合化学计量学方法可作为检测和鉴定肉毒梭菌的潜在手段。

更新日期：2021-11-09

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