Leptospirosis is an acute bacterial febrile disease affecting humans and animals in many tropical and subtropical countries. This work presents an optimization of surface-enhanced Raman spectroscopy (SERS) substrates to probe vibrational spectroscopic detail from Leptospira deoxyribonucleic acid (DNA). The pathogenic gene of LipL32 was used as a biomarker. The SERS substrates were based on a photonic crystal (PC) structure embedded with bi-metallic gold and silver nanoparticles (PC@AuAg NPs). The localized plasmonic resonance of AuAg NPs was coupled to the Raman modes of the target through SERS interaction. Prior to detection, the AuAg NPs were functionalized with chemical linkers to facilitate specific conjugation between metallic surfaces and DNA biomolecules. The immobilization and hybridization of probe DNA to their complementary target DNA (cDNA) created duplex formation for detection. The configuration was also tested with non-complementary DNA to verify detection specificity. Prominent SERS peaks were recorded, and the characteristic intensity decreased after cDNA hybridization due to less base interaction after complementary pairing. Distinct SERS behavior from the negative control test was also observed in non-complementary interaction. The configuration is highly attractive and can be potentially extended for sensitive and label-free detection of leptospiral DNA, paving the way for alternative diagnosis of leptospirosis.
KarpagamK.B.GaneshB.. “Leptospirosis: A Neglected Tropical Zoonotic Infection of Public Health Importance: An Updated Review”. Eur. J. Clin. Microbiol. Infect. Dis. 2020. 39(5): 835–846. https://doi.org/10.1007/s10096-019-03797-4
2.
BritoT.D.SilvaA.M.D.D.AbreuP.. “Pathology and Pathogenesis of Human Leptospirosis: A Commented Review”. Rev. Inst. Med. Trop. Sao Paulo. 2018. 60: e23. 10.1590/S1678-9946201860023
3.
CordoninC.TurpinM.BringartM.BascandsJ.-L., et al. “Pathogenic Leptospira and Their Animal Reservoirs: Testing Host Specificity Through Experimental Infection”. Sci. Rep. 2020. 10(1): 7239. https://doi.org/10.1038/s41598-020-64172-4
BudihalS.V.PerwezK.. “Leptospirosis Diagnosis: Competency of Various Laboratory Tests”. J. Clin. Diagn. Res. 2014. 8(1): 199–202. 10.7860/JCDR/2014/6593.3950
6.
NiloofaR.FernandoN.de SilvaN.L.KarunanayakeL., et al. “Diagnosis of Leptospirosis: Comparison Between Microscopic Agglutination Test, IgM–ELISA and IgM Rapid Immunochromatography Test”. PLoS One. 2015. 10(6): e0129236. https://doi.org/10.1371/journal.pone.0129236
7.
BackstedtB.T.BuyuktanirO.LindowJ.WunderE.A.J., et al. “Efficient Detection of Pathogenic Leptospires Using 16S Ribosomal RNA”. PLoS One. 2015. 10(6): e0128913. https://doi.org/10.1371/journal.pone.0128913
8.
FerreiraA.S.CostaP.RochaT.AmaroA., et al. “Direct Detection and Differentiation of Pathogenic Leptospira Species Using a Multi-Gene Targeted Real Time PCR Approach”. PLoS One. 2014. 9(11): e112312. https://doi.org/10.1371/journal.pone.0112312
ItohT.BijuV.IshikawaM.KikkawaY., et al. “Surface-Enhanced Resonance Raman Scattering and Background Light Emission Coupled with Plasmon of Single Ag Nanoaggregates”. J. Chem. Phys. 2006. 124(13): 134708. https://doi.org/10.1063/1.2177662
11.
PyrakE.KrajczewskiJ.KowalikA.KudelskiA., et al. “Surface Enhanced Raman Spectroscopy for DNA Biosensors: How Far Are We?”. Molecules. 2019. 24(24): 4423. https://doi.org/10.3390/molecules24244423
12.
ChenJ.LiuX.LiuZ.MaJ., et al. “Ultrasensitive SERS Biosensor for Synchronous Detection of Escherichia coli O157:H7 and Pseudomonas aeruginosa via Cecropin 1-Functionalized Magnetic Tags-Based Lateral Flow Assay”. Sens. Actuators, B. 2024. 409: 135598. https://doi.org/10.1016/j.snb.2024.135598
13.
OliveiraD.CarneiroM.C.C.G.MoreiraF.T.C.. “SERS Biosensor with Plastic Antibodies for Detection of a Cancer Biomarker Protein”. Michrochim. Acta. 2024. 191(5): 238. https://doi.org/10.1007/s00604-024-06327-y
14.
SpazianiS.QueroG.ManagòS.ZitoG., et al. “SERS Biosensor for Ultrasensitive Detection of Human Thyroglobulin”. Proc. SPIE 13008, Biophotonics in Point-of-Care III. 2024. 13008: 1300809. https://doi.org/10.1117/12.3014091
15.
YuwenL.NiJ.LiangJ.LiuX., et al. “Portable SERS Biosensor Based on Aptamer-Assisted Catalytic Hairpin Assembly Signal Amplification for Ultrasensitive Detection of Staphylococcus aureus”. Talanta. 2024. 278: 126565. https://doi.org/10.1016/j.talanta.2024.126565
16.
AlmehmadiL.M.CurleyS.M.TokranovaN.A.TenenbaumS.A., et al. “Surface Enhanced Raman Spectroscopy for Single Molecule Protein Detection”. Sci. Rep. 2019. 9(1): 12356. https://doi.org/10.1038/s41598-019-48650-y
17.
MercadalP.A.EncinaE.R.CoronadoE.A.. “Colloidal SERS Substrate for the Ultrasensitive Detection of Biotinylated Antibodies Based on Near-Field Gradient Within the Gap of Au Nanoparticle Dimers”. J. Phys. Chem. C. 2019. 123(38): 23577–23585. https://doi.org/10.1021/acs.jpcc.9b02974
18.
FuX.ChengZ.YuJ.ChooP., et al. “A SERS-Based Lateral Flow Assay Biosensor for Highly Sensitive Detection of HIV-1 DNA”. Biosens. Bioelectron. 2016. 78: 530–537. https://doi.org/10.1016/j.bios.2015.11.099
ZhuG.ChengL.LiuG.ZhuL.. “Synthesis of Gold Nanoparticle Stabilized on Silicon Nanocrystal Containing Polymer Microspheres as Effective Surface-Enhanced Raman Scattering (SERS) Substrates”. Nanomaterials. 2020. 10(8): 1501. https://doi.org/10.3390/nano10081501
21.
MakaT.ChigrinD.RomanovS.TorresC.S.. “Three Dimensional Photonic Crystals in the Visible Regime”. Prog. Electromagn. Res. 2003. 41: 307–335. https://doi.org/10.2528/PIER02010894
22.
SengwaR.J.DhatarwalP.. “Polymer Nanocomposites Comprising PMMA Matrix and ZnO, SnO2, and TiO2 Nanofillers: A Comparative Study of Structural, Optical, and Dielectric Properties for Multifunctional Technological Applications”. Opt. Mater. 2021. 113: 110837. https://doi.org/10.1016/j.optmat.2021.110837
23.
LonerganA.MurphyB.O'DwyerC.. “Photonic Stopband Tuning in Metallo-Dielectric Photonic Crystals”. ECS J. Solid State Sci. Technol. 2021. 10(8): 085001. https://doi.org/10.1149/2162-8777/ac19c5
24.
BarveenN.R.WangT.J.WangY.Y.WengC.Y., et al. “Flexible Silver-Nanoparticles/PMMA SERS Substrate Derived from Nanotips-Equipped Chemically Patterned Ferroelectric Crystals for Detecting Antibiotics on Irregular Surfaces”. Surf. Interfaces. 2023. 41: 103169. https://doi.org/10.1016/j.surfin.2023.103169
25.
WangX.Y.WuX.Y.WenX.BaiX., et al. “Composite Structure of Au Film/PMMA Grating Coated with Au Nanocubes for SERS Substrate”. Opt. Mater. 2021. 121: 111536. https://doi.org/10.1016/j.optmat.2021.111536
26.
GushikenN.K.PaganotoG.T.TemperiniM.L.A.TeixeiraF.S., et al. “Substrate for Surface-Enhanced Raman Spectroscopy Formed by Gold Nanoparticles Buried in Poly(methyl methacrylate)”. ACS Omega. 2020. 5(18): 10366–10373. https://doi.org/10.1021/acsomega.0c00133
27.
SuhailinF.H.AlwahibA.A.CheeH.Y.MustafaF.H., et al. “Fiber-Based Surface Plasmon Resonance Biosensor for Leptospira DNA Detection”. IEEE Sens. J. 2022. 22(21): 20516–20523. https://doi.org/10.1109/JSEN.2022.3207498
28.
TahrinR.A.A.AzmaN.S.KassimS.HarunN.A.. “Preparation and Properties of PMMA Nanoparticles as 3 Dimensional Photonic Crystals and Its Thin Film via Surfactant-Free Emulsion Polymerization”. AIP Conf. Proc. 2017. 1885(1): 020092. https://doi.org/10.1063/1.5002286
29.
SaidM.N.A.HasbullahN.A.RosdiM.R.H.MusaM.S., et al. “Polymerization and Applications of Poly(Methyl Methacrylate)-Graphene Oxide Nanocomposites: A Review”. ACS Omega. 2022. 7(51): 47490–47503. https://doi.org/10.1021/acsomega.2c04483
30.
TrofaM.D’AvinoG.FabianoB.VoccianteM.. “Nanoparticles Synthesis in Wet-Operating Stirred Media: Investigation on the Grinding Efficiency”. Materials. 2020. 13(19): 4281. https://doi.org/10.3390/ma13194281
31.
YewR.KaruturiS.K.LiuJ.TanH.H., et al. “Exploiting Defects in TiO(2) Inverse Opal for Enhanced Photoelectrochemical Water Splitting”. Opt. Express. 2019. 27(2): 761–773. https://doi.org/10.1364/OE.27.000761
32.
LeeH.J.LeeS.G.OhE.J.ChungH.Y., et al. “Antimicrobial Polyethyleneimine-Silver Nanoparticles in a Stable Colloidal Dispersion”. Colloids Surf., B. 2011. 88(1): 505–511. https://doi.org/10.1016/j.colsurfb.2011.07.041
33.
BaboS.FerreiraJ.L.RamosA.M.MicheluzA., et al. “Characterization and Long-Term Stability of Historical PMMA: Impact of Additives and Acrylic Sheet Industrial Production Processes”. Polymers. 2020. 12(10): 2198. https://doi.org/10.3390/polym12102198
34.
KuharN.SilS.UmapathyS.. “Potential of Raman Spectroscopic Techniques to Study Proteins”. Spectrochim. Acta, Part A. 2021. 258: 119712. https://doi.org/10.1016/j.saa.2021.119712
35.
DasG.M.WilliamR.V.DanthamV.R.LahaR.. “Study on SERS Activity of Au–Ag Bimetallic Nanostructures Synthesized Using Different Reducing Agents”. Phys. E. 2021. 129: 114656. https://doi.org/10.1016/j.physe.2021.114656
36.
FuH.BaoH.M.ZhangH.W.ZhaoQ., et al. “Quantitative Surface-Enhanced Raman Spectroscopy for Field Detections Based on Structurally Homogeneous Silver-Coated Silicon Nanocone Arrays”. ACS Omega. 2021. 6(29): 18928–18938. https://doi.org/10.1021/acsomega.1c02179
37.
KrischM.MermetA.GrimmH.ForsythV.T., et al. “Phonon Dispersion of Oriented DNA by Inelastic X-ray Scattering”. Phys. Rev. E: Stat., Nonlinear, Soft Matter Phys. 2006. 73(6): 061909. https://doi.org/10.1103/PhysRevE.73.061909
BansalJ.SinghI.BhatnagarP.K.MathurP.C.. “DNA Sequence Detection based on Raman Spectroscopy Using Single Walled Carbon Nanotube”. J. Biosci. Bioeng. 2013. 115(4): 438–441. https://doi.org/10.1016/j.jbiosc.2012.11.002
40.
HithellG.ShawD.J.DonaldsonP.M.GreethamG.M., et al. “Long-Range Vibrational Dynamics are Directed by Watson–Crick Base-Pairing in Duplex DNA”. J. Phys. Chem. B. 2016. 120(17): 4009–4018. https://doi.org/10.1021/acs.jpcb.6b02112
41.
BarhoumiA.ZhangD.TamF.HalasN.J.. “Surface-Enhanced Raman Spectroscopy of DNA”. J. Am. Chem. Soc. 2008. 130(16): 5523–5529. https://doi.org/10.1021/ja800023j
42.
DickS.BellS.E.J.AlexanderK.J.O'NeilI.A., et al. “SERS and SERRS Detection of the DNA Lesion 8-Nitroguanine: A Self-Labeling Modification”. Chem.: Eur. J. 2017. 23(44): 10663–10669. https://doi.org/10.1002/chem.201701791
43.
ZainuddinN.H.CheeH.Y.AhmadM.Z.MahdiM.A., et al. “Sensitive Leptospira DNA Detection Using Tapered Optical Fiber Sensor”. J. Biophoton. 2018. 11(8): e201700363. https://doi.org/10.1002/jbio.201700363
44.
ZainuddinN.H.CheeH.Y.RashidS.A.AhmadM.Z., et al. “Carbon Quantum Dots Functionalized Tapered Optical Fiber for Highly Sensitive and Specific Detection of Leptospira DNA”. Opt. Laser Technol. 2023. 157: 108696. https://doi.org/10.1016/j.optlastec.2022.108696
45.
HeL.J.LangletM.StambouliV.. “Role of the External NH2 Linker on the Conformation of Surface Immobilized Single Strand DNA Probes and Their SERS Detection”. Appl. Surf. Sci. 2017. 399: 702–710. https://doi.org/10.1016/j.apsusc.2016.12.131
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