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Significance: Point-of-care (POC) platforms utilizing optical biosensing strategies can achieve on-site detection of biomarkers to improve the quality of care for patients in low-resource settings., Aim: We aimed to develop a portable, multi-modal spectroscopic platform capable of performing Raman and fluorescence measurements from a single sample site., Approach: We designed the spectroscopic platform in OpticStudio using commercial optical components and built the system on a portable optical breadboard. Two excitation and collection arms were utilized to detect the two optical signals. The multi-modal functionality was validated using ratiometric Raman/fluorescence samples, and the potential utility was demonstrated using a model bioassay for cardiac troponin I., Results: The designed spectroscopic platform achieved a spectral resolution of 0.67 ± 0.2 nm across the Raman detection range (660 to 770 nm). The ratiometric Raman/fluorescence samples demonstrated no crosstalk between the two detector arms across a gradient of high molar concentrations. Testing of the model bioassay response showed that the integrated approach improved the linearity of the calibration curve from (R 2 = 0.977) for the Raman only and (R 2 = 0.972) for the fluorescence only to (R2 = 0.988) for the multi-modal approach., Conclusion: These findings demonstrate the potential impact of a multi-modal POC spectroscopic platform to improve the sensitivity and robustness necessary for biomarker detection.

Significance: Cardiac troponin I (cTnI) is a primary biomarker for diagnosis of myocardial infarction (MI). In contrast to central laboratory tests for cTnI, point-of-care (POC) testing has the advantage of providing results when the patient is first encountered, which helps high-risk patients to be treated more rapidly and low-risk patients to be released in a timely fashion. A paper fluidic platform is good for POC testing because the paper is abundant, low cost, and disposable. However, current cTnI assays on paper platforms use antibodies as the recognition element, which has limitations due to the high cost of production and antibody stability issues at the POC., Aim: To develop an aptamer-based assay on a paper strip using surface-enhanced resonance Raman spectroscopy (SERRS) for detection of cTnI in the clinically relevant range at the POC., Approach: Gold nanoparticles (AuNPs) were functionalized with a Raman reporter molecule, malachite green isothiocyanate. The functionalized AuNPs were encapsulated in a silica shell and provided a SERRS signal using a handheld Raman system with a 638-nm excitation wavelength. A primary aptamer and a secondary aptamer of cTnI were used in a sandwich assay format to bind the cTnI on a test line of a paper fluidic platform. By measuring the SERRS signal from the test line, the concentration of cTnI was quantitatively determined., Results: The aptamer-based SERRS assay on a paper strip had a detection range of 0.016 to 0.1  ng  /  ml for cTnI, had good selectivity for cTnI compared to three other markers, had good stability over 10 days, and had good performance in the more complex serum sample matrix., Conclusions: The aptamer-based SERRS assay on a paper strip has the potential to provide a sensitive, selective, stable, repeatable, and cost-effective platform for the detection of cTnI toward eventual use in diagnosis of MI at the POC.

Chip-scale optical waveguide-assisted surface-enhanced Raman spectroscopy (SERS) that used nanoparticles (NPs) was demonstrated. The Raman signals from Raman reporter (RR) molecules on NPs can be efficiently excited by the waveguide evanescent field when the molecules are in proximity to the waveguide surface. The Raman signal was enhanced by plasmon resonance due to the NPs close to the waveguide surface. The optical waveguide mode and the NP-induced field enhancement were calculated using a finite difference method (FDM). The sensing performance of the waveguide-assisted SERS device was experimentally characterized by measuring the Raman scattering from various RRs, including 4-mercaptobenzoic acid (4-MBA), 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB), and malachite green isothiocyanate (MGITC). The observed Raman spectral features were identified and assigned to the complex vibrational modes associated with different reporters. A low detection limit of 1 nM was achieved. In addition, the device sensing method was applied to the detection of the biomarker cardiac troponin I (cTnI) using an aptamer sandwich assay immobilized on the device surface. Overall, the optical waveguides integrated with SERS show a miniaturized sensing platform for the detection of small molecules and large proteins, potentially enabling multiplexed detection for clinically relevant applications. [ABSTRACT FROM AUTHOR]

A variety of innovative point-of-care (POC) solutions using Raman systems have been explored. However, the vast effort is in assay development, while studies of the characteristics required for Raman spectrometers to function in POC applications are lacking. In this study, we tested and compared the performance of eight commercial Raman spectrometers ranging in size from benchtop Raman microscopes to portable and handheld Raman spectrometers using paper fluidic cartridges, including their ability to detect cardiac troponin I and heart fatty acid binding protein, both of which are well-established biomarkers for evaluating cardiovascular health. Each spectrometer was evaluated in terms of excitation wavelength, laser characteristics, and ease of use to investigate POC utility. We found that the Raman spectrometers equipped with 780 and 785 nm laser sources exhibited a reduced background signal and provided higher sensitivity compared to those with 633 and 638 nm laser sources. Furthermore, the spectrometer equipped with the single acquisition line readout functionality showed improved performance when compared to the point scan spectrometers and allowed measurements to be made faster and easier. The portable and handheld spectrometers also showed similar detection sensitivity to the gold standard instrument. Lastly, we reduced the laser power for the spectrometer with single acquisition line readout capability to explore the system performance at a laser power that change the classification from a Class 3B laser device to a Class 3R device and found that it showed comparable performance. Overall, these findings show that portable Raman spectrometers have the potential to be used in POC settings with accuracy comparable to laboratory-grade instruments, are relatively low-cost, provide fast signal readout, are easy to use, and can facilitate access for underserved communities. Graphical Abstract This is a visual representation of the abstract. [ABSTRACT FROM AUTHOR]

Multiplexed assays are essential for the detection of biomarker panels. Differentiating signals from different biomarkers in a single test zone makes the detection more efficient. In this paper, a new method is designed for the synthesis of gap-enhanced nanoparticles (GeNPs) using Raman reporter molecules (RRM) and 6-amino-1-hexanethiol (6-AHT) as the spacer. The GeNPs show a nanometer-size gap, generate strong surface-enhanced Raman scattering (SERS) attributed to the gap, and exhibit discriminative spectral peaks. The strong Au–S bonds on both core and shell sides and the covalent bond between RRM and 6-AHT led to a stable structure, which ensured the stable SERS signal generation from the GeNPs. Using the GeNPs, a spectrally multiplexed assay for the detection of a biomarker panel is developed. The biomarker panel is composed of cardiac troponin I (cTnI), copeptin, and heart-type fatty acid-binding protein (h-FABP), which improves myocardial infarction (MI) diagnostic performance. A paper-based platform that is more amenable to point-of-care diagnostic analysis is used. The developed single biomarker assay achieves limits of detection of 0.01 ng mL−1, 0.86 ng mL−1, 0.004 ng mL−1 for cTnI, h-FABP, and copeptin in buffer solutions. The dynamic range of the assay in human serum samples also covers the clinically relevant range of the biomarkers. The cross interference in the multiplexed assay is low. These results show the strong potential of the developed GeNPs in multiplexed detection of biomarkers and the developed simple-to-use multiplexed assay in the diagnosis of MI at the point of care. [Display omitted] • GeNPs with nanometer gaps for SERS and spectral discrimination were synthesized. • Using RRMs as gap spacers ensured a high attachment of the RRM on the core surface. • 6-AHT was used with RRMs to form strong Au–S bonds on both core and shell sides. • The GeNPs were used in a simple-to-use spectrally multiplexed paper-based sensor. • Detection of cTnI, copeptin, and h-FABP, at clinical levels in serum samples. [ABSTRACT FROM AUTHOR]