Confocal Raman microspectroscopy combined with chemometrics as a discrimination method of clostridia and serotypes of <i>Clostridium botulinum</i> strains

Jin, Zhang; Jiang, Hong; Gao, Pengya; Wu, Yuan; Sun, Hui; Huang, Ying; Xu, Xuefang

doi:10.1002/jrs.6244

Cited by 4 publications

(5 citation statements)

References 61 publications

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“…Two Raman peaks around 918 cm − 1 and 1384 cm − 1 were noted only in the samples of 48 h (woman 1) and 6 h (woman 2). This observation along with the high-intensity magnitude of Raman peaks at 1156 cm − 1 and 1518 cm − 1 in our Raman spectra, lead us to assume the potential presence of BoNT/A in these two samples since these two peaks have been also detected in Clostridium botulinum B bacterium [38, 39]. However, additional experiments using more sensitive techniques such as Surface-Enhanced Raman Scattering (SERS) [51] or Fano-enhanced Raman scattering (FERS) [52] could provide complementary evidence to support this observation.…”

Section: Discussionmentioning

confidence: 71%

“…Lastly, twisting and rocking vibrations of CH 2 are noted in Raman spectrum at 870 cm −1 and 1156 cm −1 [37]. The measurements were performed in a liquid environment with an excitation wavelength at 532 nm, an oil immersion objective lens with a magnification of 100×, and an integration time of 15 s. The characteristic peaks at 918 cm −1 and 1384 cm −1 were observed in samples 48 hours (woman 1) and 6 hours (woman 2) suggesting the presence of BoNT/A [38]. Note: (I):1200 -1050 cm −1 , (II): 1300 -1250 cm −1 and (III): 1500 -1400 cm −1 .…”

Section: Raman Spectroscopy Resultsmentioning

confidence: 99%

“…Comparing the spectra in Figures 3, we noted two Raman peaks around 918 cm − 1 and 1384 cm − 1 only in the samples of 48 hours (woman 1) and 6 hours (woman 2). These peaks have been detected in Clostridium Botulinum B bacterium which can produce Botox in specific conditions [38, 39].…”

Section: Resultsmentioning

confidence: 99%

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Detection of Nontoxic BoNT/A Levels in Post-Facial Botox Injection Breastmilk using a Multi-technique Approach

Gu,

Xu,

Koviazina

et al. 2024

Preprint

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Background The use of cosmetic Botox (Botulinum Neurotoxin Type A, BoNT/A) has become increasingly prevalent. Particularly after pregnancy, postpartum depression represents one major factor motivating women to use Botox even during the lactation and breastfeeding period. Currently, there is limited understanding of the impact of Botox on lactation and the potential of its active component passing into breastmilk and affecting the infant. Methods Breastmilk samples were acquired from five women aged between 28 - 45 through a clinic in Suzhou, Jiangsu, P.R. China. Three sample sets ranged from 1 hour to 1 year after facial Botox treatments (64 U), whereas the remaining two sample sets were from women who never received Botox treatment. BoNT/A concentrations in samples were detected using standard Enzyme-Linked Immunosorbent Assay (ELISA), unreduced and reduced Western Blotting, confocal micro-Raman Spectroscopy, and Mass Spectrometry (LCMS). Findings From ELISA, breastmilk BoNT/A concentrations peaked at 33.4 pg/mL 4 days after Botox injection. BoNT/A concentrations were highest overall in the first week and around two months after injection. While non-reducing polyacrylamide gel electrophoresis (PAGE) showed a protein band of 150 kDa peaking at 48 hours, reduced SDS-PAGE detected a 100 kDa protein first peaking at 72 hours, then re-emerging after 7 days, respectively, and in line with previous observations by others. Interestingly, micro-Raman spectroscopy indicated additional Raman peaks at 6 hours and 48 hours that were not present in other breastmilk samples which were evaluated in this study. However, no clear indication of BoNT/A was detected in Mass Spectrometry (LC-MS). Interpretation The amount of BoNT/A in breastmilk peaks around 48 hours, and at 2 months after facial injection. Even over a year after injection, BoNT/A can be detected. However, all quantities of BoNT/A detected in this study are highly likely to be safe for infants. Additionally, our study suggests that alternative methods, besides ELISA, may be utilized for the rapid detection of low concentrations of BoNT/A in body fluid samples.

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Section: Discussionmentioning

confidence: 71%

Section: Raman Spectroscopy Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Detection of Nontoxic BoNT/A Levels in Post-Facial Botox Injection Breastmilk using a Multi-technique Approach

Gu,

Xu,

Koviazina

et al. 2024

Preprint

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“…Finally, the sediment was resuspended in 200 μL of 0.9% NaCl solution. To record the spectra, about 5 μL of solution was dropped onto the aluminum foil and dried at room temperature for 5 min for Raman spectra measuring ( Zhang et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

Identification of foodborne pathogenic bacteria using confocal Raman microspectroscopy and chemometrics

Zhang

Gao

et al. 2022

Front. Microbiol.

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Rapid and accurate identification of foodborne pathogenic bacteria is of great importance because they are often responsible for the majority of serious foodborne illnesses. The confocal Raman microspectroscopy (CRM) is a fast and easy-to-use method known for its effectiveness in detecting and identifying microorganisms. This study demonstrates that CRM combined with chemometrics can serve as a rapid, reliable, and efficient method for the detection and identification of foodborne pathogenic bacteria without any laborious pre-treatments. Six important foodborne pathogenic bacteria including S. flexneri, L. monocytogenes, V. cholerae, S. aureus, S. typhimurium, and C. botulinum were investigated with CRM. These pathogenic bacteria can be differentiated based on several characteristic peaks and peak intensity ratio. Principal component analysis (PCA) was used for investigating the difference of various samples and reducing the dimensionality of the dataset. Performances of some classical classifiers were compared for bacterial detection and identification including decision tree (DT), artificial neural network (ANN), and Fisher’s discriminant analysis (FDA). Correct recognition ratio (CRR), area under the receiver operating characteristic curve (ROC), cumulative gains, and lift charts were used to evaluate the performance of models. The impact of different pretreatment methods on the models was explored, and pretreatment methods include Savitzky–Golay algorithm smoothing (SG), standard normal variate (SNV), multivariate scatter correction (MSC), and Savitzky–Golay algorithm 1st Derivative (SG 1st Der). In the DT, ANN, and FDA model, FDA is more robust for overfitting problem and offers the highest accuracy. Most pretreatment methods raised the performance of the models except SNV. The results revealed that CRM coupled with chemometrics offers a powerful tool for the discrimination of foodborne pathogenic bacteria.

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“…Differential Raman spectroscopy technology was a special new Raman spectroscopy technology, which used double light source and differential spectrum processor together to effectively remove the fluorescence interference, improve the signal-to-noise ratio, with a wide detection range, simple operation, fast, non-destructive advantages, without special pretreatment. Differential Raman spectroscopy was more suitable for the detection and research of synthesis cannabinoids [12].…”

Section: Introductionmentioning

confidence: 99%

Analysis of indole and indazole amides synthetic cannabinoids by differential Raman spectroscopy based on ANN

Liu

Jiang

2022

Journal of Forensic Sciences

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A simple, rapid, accurate, and non‐destructive method was developed for the determination of cannabinoids, combining principal component analysis and multi‐layer perceptron neural network to classify indole and indazole amide synthetic cannabinoids. Under the experimental conditions of this study, 25 experimental samples were successfully classified into two categories as the final classification, which guaranteed 96% correct rate. First, the samples were manually classified and divided into two categories according to the difference in peak position and peak intensity of the differential Raman characteristic peaks at 650–540 cm−1, etc. Fisher's discriminant method (FDA) and principal component analysis (PCA) were used to analyze the experimental data. Fisher's discriminant analysis was used to formulate two classification functions to discriminate the results of manual classification, and the overall accuracy rate of classification reached 88%. Principal component analysis was used to reduce the dimensionality of the data, which could reduce the influence of redundant data on the experimental results. The original data, FDA‐processed data and PCA‐processed data, and artificial neural network algorithm (ANN‐MLP/RBF) were combined to build a classification model. In the MLP model, the classification accuracy of the original data, FDA‐processed data, and PCA‐processed data was 80%, 92%, and 96% respectively, and the overall accuracy of the sample classification was 89.33%. In the RBF model, the accuracy of sample classification was 76%, 84%, and 92% respectively, and the overall accuracy of sample classification was 84%. Differential Raman spectroscopy could be used to distinguish 25 kinds of synthetic cannabinoids, and finally, the samples were divided into two categories. The PCA + MLP model was the best for processing spectral data. Based on the perspective of multivariate data, this study demonstrated that the method could be used for rapid and non‐destructive testing of indole and indazole amide synthetic cannabinoids and that an efficient and non‐destructive classification model was obtained. This method could be used for rapid detection and inspection of drugs in the field of forensic science.

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

Cited by 4 publications

References 61 publications

Detection of Nontoxic BoNT/A Levels in Post-Facial Botox Injection Breastmilk using a Multi-technique Approach

Detection of Nontoxic BoNT/A Levels in Post-Facial Botox Injection Breastmilk using a Multi-technique Approach

Identification of foodborne pathogenic bacteria using confocal Raman microspectroscopy and chemometrics

Analysis of indole and indazole amides synthetic cannabinoids by differential Raman spectroscopy based on ANN

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