Surface-enhanced Raman spectroscopy (SERS) for the characterization of supernatants of bacterial cultures of bacterial strains causing sinusitis.

BACKGROUND: Sinusitis is defined as inflammation of the paranasal sinus mucous membrane lining caused by bacteria which usually invade the sinus by upper respiratory tract viral infections (UTI). OBJECTIVES: In the present study, Surface-enhanced Raman spectroscopy (SERS) has been applied to differentiate and characterize supernatant samples, in triplicate, of three different types of bacteria which are considered leading cause of sinusitis disease. METHODS: For this purpose, supernatant samples of three different strains of bacteria namely Staphylococcus aureus, Klebsiella pneumoniae and Enterococcus faecalis. The SERS has identified significant changes as a result of secretions of biomolecules by these bacteria in their supernatants which can be helpful to explore the potential of this technique for the identification and characterization of different strains of bacteria causing same disease. RESULTS: These differentiating characteristic SERS spectral features including 552 cm-1 (C-S-S-C bonds), 951 cm-1 (CN stretching), 1008 cm-1 (Phenylalanine), 1032 cm-1 (In plane CH bending mode Phenylalanine), 1280 cm-1, 1320 cm-1, 1329 cm-1 (Amide III band), 1368 cm-1, 1400 cm-1, 1420 cm-1 (COO-sym. stretching and CH bending), 1583 cm-1 (Tyrosine) correspond to Proteins and 1051 cm-1 (C-C, C-O, -C-OH def.) correspond to carbohydrates contents of these three different types of bacterial secretions in their respective supernatants. Furthermore, multivariate data analysis techniques like principal component analysis (PCA) and a supervised method partial least squares-discriminant analysis (PLS-DA) were found to be useful for the identification and characterization of different bacterial supernatants. CONCLUSIONS: Surface-enhanced Raman spectroscopy is proven to be a helpful approach for the characterization and discrimination of three bacterial supernatants including S. aureus, K. pneumonia and E. faecalis.

Surface-enhanced Raman spectroscopy for the characterization of pellets of biofilm forming bacterial strains of Staphylococcus epidermidis.

BACKGROUND: Surface-enhanced Raman spectroscopy (SERS) is an effective tool for identifying biofilm forming bacterial strains. Biofilm forming bacteria are considered a major issue in the health sector because they have strong resistance against antibiotics. Staphylococcus epidermidis is commonly present on intravascular devices and prosthetic joints, catheters and wounds. OBJECTIVES: To identify and characterize biofilm forming and non-biofilm forming bacterial strains, surface- enhanced Raman spectroscopy with principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) were used. METHODS: Surface-enhanced Raman spectroscopy (SERS) with silver nanoparticles were employed for the analysis and characterization of biofilm forming bacterial strains. SERS is used to differentiate between non biofilm forming (five samples), medium biofilm forming (five samples) and strong biofilm forming (five samples) bacterial strains by applying silver nanoparticles (AgNPs) as SERS substrate. Principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA) were used to discriminate between non, medium and strong biofilm ability of bacterial strains. RESULTS: Principal component analysis (PCA) and Partial least square discriminant analysis (PLS-DA) have been used to identify the biochemical differences in the form of SERS features which can be used to differentiate between biofilm forming and non-biofilm forming bacterial strains. PLS-DA provides successful differentiation and classification of these different strains with 94.5% specificity, 96% sensitivity and 89% area under the curve (AUC). CONCLUSIONS: Surface-enhanced Raman spectroscopy can be utilized to differentiate between non, medium and strong biofilm forming bacterial strains.

Surface-enhanced Raman spectroscopy (SERS) for monitoring colistin-resistant and susceptible E. coli strains.

The emergence of drug-resistant bacteria is a precarious global health concern. In this study, surface-enhanced Raman spectroscopy (SERS) is used to characterize colistin-resistant and susceptible E. coli strains based on their distinguished SERS spectral features for the development of rapid and cost-effective detection and differentiation methods. For this purpose, three colistin-resistant and three colistin susceptible E. coli strains were analyzed by comparing their SERS spectral signatures. Moreover, multivariate data analysis techniques including Principal component analysis (PCA) and Partial Least Squares-Discriminant Analysis (PLS-DA) were used to examine the SERS spectral data of colistin-resistant and susceptible strains. PCA technique was employed for differentiating colistin susceptible and resistant E.coli strains due to alteration in biochemical compositions of the bacterial cell. PLS-DA is employed on SERS spectral data sets for discrimination of these resistant and susceptible E. coli strains with 100% specificity, 100% accuracy, 99.8% sensitivity, and 86% area under receiver operating characteristics (AUROC) curve.

Quantitative analysis of solid dosage forms of Losartan potassium by Raman spectroscopy.

Raman spectroscopy is an outstanding analytical tool increasingly utilized in the pharmaceutical field for the solid-state pharmaceutical drug analysis. In current study, the potential of Raman spectroscopy has been investigated for qualitative and quantitative analysis of solid dosage form of Losartan potassium. For this purpose, different solid dosage forms/concentrations of losartan potassium were prepared to compensate the commercially available pharmaceutical drug formulations and their Raman spectral data showed a gradual change in the specific Raman spectral features associated with the active pharmaceutical ingredient (API) of Losartan potassium as a function of change in the concentration. The Raman spectral data was analyzed by using Principal Component Analysis (PCA) for the classification of different spectral data sets of different concentrations of drug. Moreover, partial least square regression (PLSR) analysis was performed for monitoring the quantitative relation among different concentrations of Losartan potassium API and spectral data by constructing a predictive model. From the model, the value of root mean square error of calibration (RMSEC) and root mean square error of prediction (RMSEP) were observed to be 0.38 and 2.98 respectively and the value of goodness of fit was found to be 0.99. Furthermore, the quantity of unknown/blind sample of Losartan potassium formulation was also estimated by using PLSR model. From these results, it is demonstrated that Raman spectroscopy can be considered to be used for quick and reliable quantitative analysis of pharmaceutical solids.

Surface-enhanced Raman spectroscopy for comparison of serum samples of typhoid and tuberculosis patients of different stages.

BACKGROUND: Surface-enhanced Raman spectroscopy (SERS) is a reliable tool for the identification and differentiation of two different human pathological conditions sharing the same symptomology, typhoid and tuberculosis (TB). OBJECTIVES: To explore the potential of surface-enhanced Raman spectroscopy for differentiation of two different diseases showing the same symptoms and analysis by principal component analysis (PCA) and partial least square discriminate analysis (PLS-DA). METHODS: Serum samples of clinically diagnosed typhoid and tuberculosis infected individuals were analyzed and differentiated by SERS using silver nanoparticles (Ag NPs) as a SERS substrate. For this purpose, the collected serum samples were analyzed under the SERS instrument and unique SERS spectra of typhoid and tuberculosis were compared showing notable spectral differences in protein, lipid and carbohydrates features. Different stages of the diseased class of typhoid (Early acute and late acute stage) and tuberculosis (Pulmonary and extra-pulmonary stage) were compared with each other and with healthy human serum samples, which were significantly separated. Moreover, SERS data was analyzed using multivariate data analysis techniques including principal component analysis (PCA) and partial least square discriminate analysis (PLS-DA) and differences were so prominent to observe. RESULTS: SERS Spectral data of typhoid and tuberculosis showed clear differences and were significantly separated using PCA. SERS spectral data of both stages of typhoid and tuberculosis were separated according to 1st principle component. Moreover, by analyzing data using partial least square discriminate analysis, differentiation of two disease classes were considered more valid with a 100% value of sensitivity, specificity and accuracy. CONCLUSION: SERS can be employed for identification and comparison of two different human pathological conditions sharing same symptomology.

Surface-enhanced Raman spectral analysis for comparison of PCR products of hepatitis B and hepatitis C.

BACKGROUND: Surface-enhanced Raman spectroscopy is a reliable tool for identification and differentiation of two diseases showing similar symptoms, hepatitis B (HBV) and hepatitis C (HCV). OBJECTIVES: To develop a polymerase chain reaction technique (PCR) based SERS technique for differentiation of two human pathological conditions sharing the same symptoms using multivariate data analysis techniques e.g. principle component analysis (PCA) and partial least square discriminate analysis (PLS-DA). METHODS: PCR products of HBV and HCV were differentiated by SERS using silver nanoparticles (AgNPs) as a SERS substrate. For this analysis, PCR products of both the diseases with predetermined viral loads were collected and analyzed under SERS instrument and unique SERS spectra of HBV and HCV was compared showing many differences at various points. Diseased classes of HBV and HCV and their negative control classes (viral load less than 1) were compared. PCR products of true healthy DNA and RNA were also compared, which were significantly separated. Moreover, SERS data was analyzed using multivariate data analysis techniques including principle component analysis (PCA) and partial least square discriminate analysis (PLS-DA) and differences were so prominent to observe. RESULTS: SERS spectral data of HBV and HCV showed clear differences and were significantly separated using PCA. Negative control samples of both disorders and their true healthy samples of DNA and RNA were separated according to 1st principle component. By analyzing data using partial least square discriminate analysis, differentiation of two disease classes was considered more valid with sensitivity, specificity and accuracy value of 96%, 94% and 98% respectively. Value of area under curve (AUROC) was 0.7527. CONCLUSION: SERS can be employed for identification and comparison of two human pathological conditions sharing the same symptomology.