Characterization of functionalized support layer performance for surface plasmon resonance sensing platform for viral protein detection / Sharifah Norsyahindah Syed Nor
In the event of virus outbreak, rapid detection of viral infection is crucial for proper patient management and halting the progress of the disease into a pandemic. The current gold standard of virus detection is the reverse transcription-polymerase chain reaction (RT-PCR), which is time consuming a...
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| Format: | Thesis |
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2024
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| Online Access: | http://studentsrepo.um.edu.my/15387/2/Sharifah_Syahindah.pdf http://studentsrepo.um.edu.my/15387/1/Sharifah_Norsyahindah.pdf http://studentsrepo.um.edu.my/15387/ |
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| Summary: | In the event of virus outbreak, rapid detection of viral infection is crucial for proper patient management and halting the progress of the disease into a pandemic. The current gold standard of virus detection is the reverse transcription-polymerase chain reaction (RT-PCR), which is time consuming and depends on relative RNA levels to determine the infection status of patients. Investigators have taken interest in surface plasmon resonance (SPR) biosensing for rapid viral detection. Therefore, the objective of this study is to develop a rapid, accurate, and ultra-sensitive SPR sensing platform for virus detection in minimum concentration. Methodologically, the reflectance of the 50 nm gold (Au) coated BK7 prism (Au/BK7 prism) was determined as reference. Next, Au sensing surface was integrated with self-assembled monolayer (SAM) those are 11-Mercaptoundecanoic acid (11-MUA), 3-Mercaptopropionic acid (3-MPA), and mix thiol (11-MUA:3-MPA in 1:10 molar ratio) in addition to transition-metal dichalcogenide (TMDC) carboxyl molybdenum disulfide (MoS2-COOH) layer to enhance the molecular adsorption that consequently enhances the sensitivity of the sensor. The support layers were characterized using Raman spectroscopy, Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM) before and after deposition onto the Au surface to study its morphology. Then, SPR measurements were performed for the four configurations (11-MUA/Au/BK7 prism, 3-MPA/Au/BK7 prism, mix thiol/Au/BK7 prism, and MoS2-COOH/Au/BK7 prism) to observe the changes to the SPR curve that proves the interaction of the support layers with Au surface. Consequently, each configuration undergoes refractive index (RI) sensitivity characterization with ethanol at different concentrations (0.01 M, 1 M, 5 M, 10 M, and 16.98 M) as samples to determine the capability of the platform to detect RI changes of the sensing medium. Next, the three most sensitive configurations are validated with testing using severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein (S protein) of various concentrations (10, 20, 30, 40, 50, 60, and 70 ng/mL) with RI variation (1.335, 1.338, 1.341, 1.343, 1.345, 1.347, and 1.350) respectively. Performance parameters such as sensitivity, full width half maximum (FWHM), detection accuracy (DA), figure of merit (FoM), quality factor (QF), and limit of detection (LoD) of the three selected configurations of different sensing element for the detection of SARS-CoV-2 S protein in varying concentration are evaluated and compared. Result shows MoS2-COOH/Au/BK7 prism exhibited the highest sensitivity (203.55 °/RIU) compared to 11-MUA/Au/BK7 prism (89.89 °/RIU) and 3-MPA/Au/BK7 prism (108.90 °/RIU). The LoD for 11-MUA/Au/BK7 prism, 3-MPA/Au/BK7 prism, MoS2-COOH/Au/BK7 prism is 6.55 ng/mL, 6.20 ng/mL, and 5.25 ng/mL respectively. Generally, all three configurations establish good linearity, excellent sensitivity, and LoD in ng/mL range for the detection of SARS-CoV-2 S protein. The findings from this study can be used to aid the future development of non-invasive, rapid, and ultra-sensitive SPR sensing platform for detection of virus infection in the minimum concentration.
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