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Surface-enhanced raman spectroscopy with machine learning in non-invasive detection of dengue-NS1 fingerprint

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Afaf Rozan Mohd Radzol

Centre for Electrical Engineering Studies, Universiti Teknologi MARA, Cawangan Pulau Pinang, Pulau Pinang, Malaysia.

Khuan Y Lee

Center of System Studies, School of Electrical Engineering, Collage of Engineering, Universiti Teknologi MARA, Selangor, Malaysia.

Peng Shyan Wong

Infectious Disease Unit, Penang General Hospital, Georgetown, Pulau Pinang, Malaysia

Irene Looi

Clinical Research Centre, Seberang Jaya Hospital, Seberang Jaya, Perai, Pulau Pinang, Malaysia

Wahidah Mansor

Center of System Studies, School of Electrical Engineering, Collage of Engineering, Universiti Teknologi MARA, Selangor, Malaysia

Abstract
The surface-enhanced Raman spectroscopy (SERS) method exploits the plasmonic effect of nano-sized metallic materials to intensify the Raman scattering of the monochromatic light of analyte molecules. This promotes the sensitivity and specificity of the Raman spectroscopy analysis method. This study integrated SERS with machine learning (ML) to detect dengue fever, a disease infecting more than 40% of the world’s population. Non-structural protein 1 (NS1), detected in the sera of infected dengue patients during the early infection stage, is currently recognised as a biomarker for the early diagnosis of DF. However, no attempts have been made to detect NS1 in the salivary Raman spectra. Given this situation, this study delves into the potential of SERS as an early, non-invasive DF detection technique using salivary NS1. The SERS spectra of saliva samples (n=289) were collected and subsequently classified as positive and negative for DF, using principal component analysis (PCA) integrated with support vector machine (SVM) models. The PCA-SVM model's performance was benchmarked against two clinical diagnostic NS1-enzyme-linked immunosorbent assay (ELISA) tests recommended by the World Health Organization (WHO). The PCA-SVM model outperformed both tests regarding radial basis function kernel (RBF) and cumulative percent variance (CPV; 83.22% accuracy, 88.27% sensitivity, 78.13% specificity). It is encouraging that the sensitivity level of the PCA-SVM model is above the benchmark set by the saliva-based NS1-ELISA tests proposed by the WHO, demonstrating the potential of SERS for the non-invasive detection of DF.

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Keyword: SERS, Machine Learning, Dengue, NS1

DOI: https://doi.org/10.24191/esteem.v20iMarch.616.g534

References:

[1]  W. H. Organization, Dengue guidelines for diagnosis, treatment, prevention and control: new edition, World Health Organization, 2009.

[2]  S. Alcon, A. Talarmin, M. Debruyne, A. Falconar, V. Deubel, and M. Flamand, “Enzyme-linked immunosorbent assay specific to Dengue virus type 1 nonstructural protein NS1 reveals circulation of the antigen in the blood during the acute phase of disease in patients experiencing primary or secondary infections,” J Clin Microbiol, vol. 40, no. 2, pp. 376–381, 2002. Available: https://doi.org/10.1128/jcm.40.02.376-381.2002

[3] D. H. Libraty et al., “High circulating levels of the dengue virus nonstructural protein NS1 early in dengue illness correlate with the development of dengue hemorrhagic fever,” J Infect Dis, vol. 186, no. 8, pp. 1165–1168, 2002. Available: https://doi.org/10.1086/343813

[4] K. L. Anders et al., “An evaluation of dried blood spots and oral swabs as alternative specimens for the diagnosis of dengue and screening for past dengue virus exposure,” Am J Trop Med Hyg, vol. 87, no. 1, p. 165, 2012. Available: https://doi.org/10.4269/ajtmh.2012.11-0713

[5] I. Gutsche et al., “Secreted dengue virus nonstructural protein NS1 is an atypical barrel-shaped high-density lipoprotein,” Proceedings of the National Academy of Sciences, vol. 108, no. 19, pp. 8003–8008, 2011. Available: https://doi.org/10.1073/pnas.1017338108

[6] D. A. Muller et al., “Structure of the dengue virus glycoprotein non-structural protein 1 by electron microscopy and single-particle analysis,” Journal of General Virology, vol. 93, no. 4, pp. 771–779, 2012. Available: https://doi.org/10.1099/vir.0.039321-0

[7] D. L. Akey, W. C. Brown, J. Jose, R. J. Kuhn, and J. L. Smith, “Structure?guided insights on the role of NS1 in flavivirus infection,” Bioessays, vol. 37, no. 5, pp. 489–494, 2015. Available: https://doi.org/10.1002/bies.201400182

[8] V. Deubel, R. M. Kinney, and D. W. Trent, “Nucleotide sequence and deduced amino acid sequence of the nonstructural proteins of dengue type 2 virus, Jamaica genotype: comparative analysis of the full-length genome,” Virology, vol. 165, no. 1, pp. 234–244, 1988. Available: https://doi.org/10.1016/0042-6822(88)90677-0

[9] W. H. Attatippaholkun, M. K. Attatippaholkun, A. Nisalak, D. W. Vaughn, and B. L. Innis, “Nucleotide sequence and deduced amino acid sequence of the nonstructural proteins of dengue type 3 virus, Bangkok genotype.,” Southeast Asian J Trop Med Public Health, vol. 29, no. 2, pp. 361–366, 1998.

[10] P. Y. Yang, I. Kautner, C. L. Koh, and S. K. Lam, “Nucleotide and deduced amino acid sequences of genes encoding the structural and nonstructural NS1 proteins of a dengue-2 virus isolated in China,” Virus Genes, vol. 8, no. 1, pp. 71–74, 1994. Available: https://doi.org/10.1007/BF01703603

[11] P. W. Mason, P. C. McAda, T. L. Mason, and M. J. Fournier, “Sequence of the dengue-1 virus genome in the region encoding the three structural proteins and the major nonstructural protein NS1,” Virology, vol. 161, no. 1, pp. 262–267, 1987. Available: https://doi.org/10.1016/0042-6822(87)90196-6

[12] B. J. Blitvich, D. Scanlon, B. J. Shiell, J. S. Mackenzie, K. Pham, and R. A. Hall, “Determination of the intramolecular disulfide bond arrangement and biochemical identification of the glycosylation sites of the nonstructural protein NS1 of Murray Valley encephalitis virus.,” J Gen Virol, vol. 82, no. Pt 9, pp. 2251–2256, 2001. Available: https://doi.org/10.1099/0022-1317-82-9-2251

[13] T. P. Wallis, C. Y. Huang, S. B. Nimkar, P. R. Young, and J. J. Gorman, “Determination of the disulfide bond arrangement of dengue virus NS1 protein,” Journal of Biological Chemistry, vol. 279, no. 20, pp. 20729–20741, 2004. Available: https://doi.org/10.1074/jbc.M312907200

[14] M. Flamand, F. Megret, M. Mathieu, J. Lepault, F. A. Rey, and V. Deubel, “Dengue virus type 1 nonstructural glycoprotein NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion,” J Virol, vol. 73, no. 7, pp. 6104–6110, 1999. Available: https://doi.org/10.1128/jvi.73.7.6104-6110.1999

[15] D. L. Akey et al., “Flavivirus NS1 Structures Reveal Surfaces for Associations with Membranes and the Immune System,” Science (1979), vol. 343, no. 6173, pp. 881–885, 2014. Available: https://www.science.org/doi/10.1126/science.1247749

[16] P. Avirutnan et al., “Vascular Leakage in Severe Dengue Virus Infections: A Potential Role for the Nonstructural Viral Protein NS1 and Complement,” J Infect Dis, vol. 193, pp. 1078–1088, 2006. Available: https://doi.org/10.1086/500949

[17] J. M. Mackenzie, M. K. Jones, and P. R. Young, “Immunolocalization of the dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication.,” Virology, vol. 220, no. 1, pp. 232–240, 1996. Available: https://doi.org/10.1006/viro.1996.0307

[18] M. Fleischmann, P. J. Hendra, and A. J. McQuillan, “Raman Spectra of Pyridine Adsorbed at a Silver Electrode,” Chem Phys Lett, vol. 26, no. 2, 1974. Available: https://doi.org/10.1016/0009-2614(74)85388-1

[19] K. Kneipp et al., “Single molecule detection using surface-enhanced Raman scattering (SERS),” Phys Rev Lett, vol. 78, no. 9, pp. 1667, 1997. Available:  https://doi.org/10.1103/PhysRevLett.78.1667

[20] S. Nie and S. R. Emory, “Probing Single Molecules and Single Nanoparticles by Surface-Enhanced Raman Scattering,” Science (1979), pp. 1102–1106, 1997. Available: https://www.science.org/doi/10.1126/science.275.5303.1102

[21] Feng et al., “Surface-enhanced Raman spectroscopy of saliva proteins for the noninvasive differentiation of benign and malignant breast tumors,” Int J Nanomedicine, pp. 537–547, 2015.

[22] J. C. Y. Kah et al., “Early diagnosis of oral cancer based on the surface plasmon resonance of gold nanoparticles,” Int J Nanomedicine, vol. 2, no. 4, pp. 785, 2007. Available: https://doi.org/10.2147/IJN.S71811

[23] C. Anyu et al., “Detecting Narcotic Usage Using Surface-Enhanced Raman Spectroscopy on Saliva Samples,” in World Congress on Medical Physics and Biomedical Engineering September 7-12, 2009, pp. 71. Available: https://doi.org/10.1007/978-3-642-03885-3_20

[24] Y. Wang et al., “A feasibility study of early detection of lung cancer by saliva test using Surface Enhanced Raman scattering,” in 2012 5th International Conference on BioMedical Engineering and Informatics, IEEE, 2012, pp. 135–139. Available: 10.1109/BMEI.2012.6513160

[25] E. Widjaja, W. Zheng, and Z. Huang, “Classification of colonic tissues using near-infrared Raman spectroscopy and support vector machines,” Int J Oncol, vol. 32, no. 3, pp. 653–662, 2008.

[26] A. R. M. Radzol, K. Y. Lee, W. Mansor, and F. M. T. Tawi, “Signal processing for raman spectra for disease detection,” Int. J. Pharm. Pharm. Sci, vol. 8, no. 6, pp. 4–10, 2016.

[27] M. Saleem, M. Bilal, S. Anwar, A. Rehman, and M. Ahmed, “Optical diagnosis of dengue virus infection in human blood serum using Raman spectroscopy,” Laser Phys Lett, vol. 10, no. 3, p. 035602, 2013. Available: https://iopscience.iop.org/article/10.1088/1612-2011/10/3/035602

[28] M. Bilal et al., “Raman spectroscopy based discrimination of NS1 positive and negative dengue virus infected serum,” Laser Phys Lett, vol. 13, no. 9, pp. 095603, 2016. Available: https://iopscience.iop.org/article/10.1088/1612-2011/13/9/095603

[29] S. Khan et al., “Raman spectroscopic analysis of dengue virus infection in human blood sera,” Optik (Stuttg), vol. 127, no. 4, pp. 2086–2088, 2016. Available: https://doi.org/10.1016/j.ijleo.2015.11.060

[30] S. Khan, R. Ullah, A. Khan, N. Wahab, M. Bilal, and M. Ahmed, “Analysis of dengue infection based on Raman spectroscopy and support vector machine (SVM),” Biomed Opt Express, vol. 7, no. 6, pp. 2249–2256, 2016. Available: https://doi.org/10.1364/BOE.7.002249

[31] S. Khan et al., “Random forest-based evaluation of Raman spectroscopy for dengue fever analysis,” Appl Spectrosc, vol. 71, no. 9, pp. 2111–2117, 2017.

[32] A. Amin, N. Ghouri, S. Ali, M. Ahmed, M. Saleem, and J. Qazi, “Identification of new spectral signatures associated with dengue virus infected sera,” Journal of Raman Spectroscopy, vol. 48, no. 5, pp. 705–710, 2017. Available: https://doi.org/10.1177/000370281769557

[33] T. Mahmood et al., “Raman spectral analysis for rapid screening of dengue infection,” Spectrochim Acta A Mol Biomol Spectrosc, vol. 200, pp. 136–142, 2018. Available: https://doi.org/10.1016/j.saa.2018.04.018

[34] S. K. Gahlaut, D. Savargaonkar, C. Sharan, S. Yadav, P. Mishra, and J. P. Singh, “SERS platform for dengue diagnosis from clinical samples employing a hand held Raman spectrometer,” Anal Chem, vol. 92, no. 3, pp. 2527–2534, 2020. Available: https://doi.org/10.1021/acs.analchem.9b04129

[35] I. T. Jolliffe, “Principal component analysis: a beginner’s guide—I. Introduction and application,” Weather, vol. 45, no. 10, pp. 375–382, 1990. Available: https://doi.org/10.1002/j.1477-8696.1990.tb05558.x

[36] V. Vapnik, “Support-vector networks,” Mach Learn, vol. 20, pp. 273–297, 1995. Available: https://doi.org/10.1007/BF00994018

[37] M. Navazesh, “Methods for collecting saliva,” Ann N Y Acad Sci, vol. 694, no. 1, pp. 72–77, 1993. Available: https://doi.org/10.1111/j.1749-6632.1993.tb18343.x

[38] N. H. Othman, K. Y. Lee, A. R. M. Radzol, and W. Mansor, “Optimized Raman Setting of Objective Lens, Laser Power and Integration Time for High and Low Concentration of Nonstructural Protein 1,” in EMBEC & NBC 2017: Joint Conference of the European Medical and Biological Engineering Conference (EMBEC) and the Nordic-Baltic Conference on Biomedical Engineering and Medical Physics (NBC), Tampere, Finland, June 2017, Springer, 2018, pp. 482–485. Available: https://doi.org/10.1007/978-981-10-5122-7_121

[39] A. R. M. Radzol, K. Y. Lee, W. Mansor, and A. Azman, “Optimization of Savitzky-Golay smoothing filter for salivary surface enhanced Raman spectra of non structural protein 1,” in TENCON 2014-2014 IEEE Region 10 Conference, IEEE, 2014, pp. 1–6. Available: https://ieeexplore.ieee.org/document/7022409

[40] A. R. Radzol, K. Y. Lee, W. Mansor, and N. Saadun, “Optimized Automated Baseline Correction for NS1 Adulterated Salivary Raman Spectra,” Adv Sci Lett, vol. 24, no. 2, pp. 1182–1186, 2018. Available: https://doi.org/10.1166/asl.2018.10712

[41] R. B. Cattell, “The scree test for the number of factors,” Multivariate Behav Res, vol. 1, no. 2, pp. 245–276, 1966. Available: https://doi.org/10.1207/s15327906mbr0102_10

[42] S. Han, C. Qubo, and H. Meng, “Parameter selection in SVM with RBF kernel function,” in World Automation Congress 2012, IEEE, 2012, pp. 1–4.