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1.
Discov Nano ; 19(1): 151, 2024 Sep 17.
Artículo en Inglés | MEDLINE | ID: mdl-39289310

RESUMEN

With the size of the aging population increasing worldwide, the effective diagnosis and treatment of neurodegenerative diseases (NDDs) has become more important. Two-dimensional (2D) materials offer specific advantages for the diagnosis and treatment of NDDs due to their high sensitivity, selectivity, stability, and biocompatibility, as well as their excellent physical and chemical characteristics. As such, 2D materials offer a promising avenue for the development of highly sensitive, selective, and biocompatible theragnostics. This review provides an interdisciplinary overview of advanced 2D materials and their use in biosensors, drug delivery, and tissue engineering/regenerative medicine for the diagnosis and/or treatment of NDDs. The development of 2D material-based biosensors has enabled the early detection and monitoring of NDDs via the precise detection of biomarkers or biological changes, while 2D material-based drug delivery systems offer the targeted and controlled release of therapeutics to the brain, crossing the blood-brain barrier and enhancing treatment effectiveness. In addition, when used in tissue engineering and regenerative medicine, 2D materials facilitate cell growth, differentiation, and tissue regeneration to restore neuronal functions and repair damaged neural networks. Overall, 2D materials show great promise for use in the advanced treatment of NDDs, thus improving the quality of life for patients in an aging population.

2.
Biosensors (Basel) ; 14(4)2024 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-38667190

RESUMEN

Controlling the progression of contagious diseases is crucial for public health management, emphasizing the importance of early viral infection diagnosis. In response, lateral flow assays (LFAs) have been successfully utilized in point-of-care (POC) testing, emerging as a viable alternative to more traditional diagnostic methods. Recent advancements in virus detection have primarily leveraged methods such as reverse transcription-polymerase chain reaction (RT-PCR), reverse transcription-loop-mediated isothermal amplification (RT-LAMP), and the enzyme-linked immunosorbent assay (ELISA). Despite their proven effectiveness, these conventional techniques are often expensive, require specialized expertise, and consume a significant amount of time. In contrast, LFAs utilize nanomaterial-based optical sensing technologies, including colorimetric, fluorescence, and surface-enhanced Raman scattering (SERS), offering quick, straightforward analyses with minimal training and infrastructure requirements for detecting viral proteins in biological samples. This review describes the composition and mechanism of and recent advancements in LFAs for viral protein detection, categorizing them into colorimetric, fluorescent, and SERS-based techniques. Despite significant progress, developing a simple, stable, highly sensitive, and selective LFA system remains a formidable challenge. Nevertheless, an advanced LFA system promises not only to enhance clinical diagnostics but also to extend its utility to environmental monitoring and beyond, demonstrating its potential to revolutionize both healthcare and environmental safety.


Asunto(s)
Técnicas Biosensibles , Nanoestructuras , Espectrometría Raman , Proteínas Virales , Humanos , Técnicas Biosensibles/métodos , Colorimetría , Nanoestructuras/química , Pruebas en el Punto de Atención , Proteínas Virales/análisis
3.
Biosensors (Basel) ; 14(1)2024 Jan 02.
Artículo en Inglés | MEDLINE | ID: mdl-38248403

RESUMEN

In the context of virus outbreaks, the need for early and accurate diagnosis has become increasingly urgent. In addition to being crucial for effective disease control, timely and precise detection of viral infections is also necessary for the implementation of essential public health measures, especially during pandemics. Among these measures, point-of-care testing (POCT) stands out as a powerful approach with the potential to revolutionize the landscape of viral diagnosis. In this study, we developed a one-pot clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a-based viral DNA detection system tailored for POCT; this method utilizes multi-enzyme-modified Au@Fe3O4 nanoparticles. As an alternative to nucleic acid amplification, our method uses single-stranded DNA elongation to facilitate multi-enzyme modification; this guarantees heightened sensitivity and expedites the diagnostic process. We achieved a satisfactory limit of detection of 0.25 nM, demonstrating the remarkable sensitivity of the method without the need for sophisticated equipment. The incorporation of Au@Fe3O4 magnetic nanoparticles facilitates sample separation, further streamlining the workflow and reinforcing the simplicity of our method. This integrated approach offers a practical solution for sensitive viral DNA detection in POCT scenarios, advancing the field of rapid and accurate diagnostics.


Asunto(s)
Sistemas CRISPR-Cas , Nanopartículas , ADN de Cadena Simple , ADN Viral , Pandemias
4.
Adv Sci (Weinh) ; 11(4): e2305371, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38036423

RESUMEN

Biohybrid robots have been developed for biomedical applications and industrial robotics. However, the biohybrid robots have limitations to be applied in neurodegenerative disease research due to the absence of a central nervous system. In addition, the organoids-on-a-chip has not yet been able to replicate the physiological function of muscle movement in the human motor system, which is essential for evaluating the accuracy of the drugs used for treating neurodegenerative diseases. Here, a human motor system-based biohybrid robot-on-a-chip composed of a brain organoid, multi-motor neuron spheroids, and muscle bundle on solid substrateis proposed to evaluate the drug effect on neurodegenerative diseases for the first time. The electrophysiological signals from the cerebral organoid induced the muscle bundle movement through motor neuron spheroids. To evaluate the drug effect on Parkinson's disease (PD), a patient-derived midbrain organoid is generated and incorporated into a biohybrid robot-on-a-chip. The drug effect on PD is successfully evaluated by measuring muscle bundle movement. The muscle bundle movement of PD patient-derived midbrain organoid-based biohybrid robot-on-a-chip is increased from 4.5 ± 0.99 µm to 18.67 ± 2.25 µm in response to levodopa. The proposed human motor system-based biohybrid robot-on-a-chip can serve as a standard biohybrid robot model for drug evaluation.


Asunto(s)
Enfermedades Neurodegenerativas , Enfermedad de Parkinson , Robótica , Humanos , Enfermedades Neurodegenerativas/tratamiento farmacológico , Evaluación de Medicamentos , Dispositivos Laboratorio en un Chip
5.
Biosensors (Basel) ; 13(9)2023 Sep 19.
Artículo en Inglés | MEDLINE | ID: mdl-37754127

RESUMEN

Neurotransmitters are chemical compounds released by nerve cells, including neurons, astrocytes, and oligodendrocytes, that play an essential role in the transmission of signals in living organisms, particularly in the central nervous system, and they also perform roles in realizing the function and maintaining the state of each organ in the body. The dysregulation of neurotransmitters can cause neurological disorders. This highlights the significance of precise neurotransmitter monitoring to allow early diagnosis and treatment. This review provides a complete multidisciplinary examination of electrochemical biosensors integrating nanomaterials and nanotechnologies in order to achieve the accurate detection and monitoring of neurotransmitters. We introduce extensively researched neurotransmitters and their respective functions in biological beings. Subsequently, electrochemical biosensors are classified based on methodologies employed for direct detection, encompassing the recently documented cell-based electrochemical monitoring systems. These methods involve the detection of neurotransmitters in neuronal cells in vitro, the identification of neurotransmitters emitted by stem cells, and the in vivo monitoring of neurotransmitters. The incorporation of nanomaterials and nanotechnologies into electrochemical biosensors has the potential to assist in the timely detection and management of neurological disorders. This study provides significant insights for researchers and clinicians regarding precise neurotransmitter monitoring and its implications regarding numerous biological applications.

6.
Biosens Bioelectron ; 237: 115474, 2023 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-37364302

RESUMEN

Microcystis aeruginosa (M. aeruginosa) cause the eutrophication of lakes and rivers. To effectively control the overgrowth of M. aeruginosa, a suitable measurement method should be required in the aquatic fields. To address this, we developed a field-ready cyanobacterial pretreatment device and an electrochemical clustered regularly interspaced short palindromic repeats (EC-CRISPR) biosensor. The cyanobacterial pretreatment device consists of a syringe, glass bead, and graphene oxide (GO) bead. Then, the M. aeruginosa dissolved in the freshwater sample was added to fabricated filter. After filtration, the purified gene was loaded onto a CRISPR-based electrochemical biosensor chip to detect M. aeruginosa gene fragments. The biosensor was composed of CRISPR/Cpf1 protein conjugated with MXene on an Au microgap electrode (AuMGE) integrated into a printed circuit board (PCB). This AuMGE/PCB system maximizes the signal-to-noise ratio, which controls the working and counter electrode areas requiring only 3 µL samples to obtain high reliability. Using the extracted M. aeruginosa gene with a pre-treatment filter, the CRISPR biosensor showed a limit of detection of 0.089 pg/µl in fresh water. Moreover, selectivity test and matrix condition test carried out using the EC-CRISPR biosensor. These handheld pre-treatment kit and biosensors can enable field-ready detection of CyanoHABs.

7.
Biosensors (Basel) ; 13(2)2023 Jan 21.
Artículo en Inglés | MEDLINE | ID: mdl-36831937

RESUMEN

Proteolytic enzymes are one of the important biomarkers that enable the early diagnosis of several diseases, such as cancers. A specific proteolytic enzyme selectively degrades a certain sequence of a polypeptide. Therefore, a particular proteolytic enzyme can be selectively quantified by changing detectable signals causing degradation of the peptide chain. In addition, by combining polypeptides with various functional nanomaterials, proteolytic enzymes can be measured more sensitively and rapidly. In this paper, proteolytic enzymes that can be measured using a polypeptide degradation method are reviewed and recently studied functional nanomaterials-based proteolytic biosensors are discussed. We anticipate that the proteolytic nanobiosensors addressed in this review will provide valuable information on physiological changes from a cellular level for individual and early diagnosis.


Asunto(s)
Técnicas Biosensibles , Nanoestructuras , Péptido Hidrolasas , Biomarcadores , Péptidos , Técnicas Biosensibles/métodos
8.
Biosensors (Basel) ; 13(2)2023 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-36832055

RESUMEN

Hydrogen peroxide (H2O2) has been a fascinating target in various chemical, biological, clinical, and industrial fields. Several types of fluorescent protein-stabilized gold nanoclusters (protein-AuNCs) have been developed for sensitive and easy detection of H2O2. However, its low sensitivity makes is difficult to measure negligible concentrations of H2O2. Therefore, to overcome this limitation, we developed a horseradish peroxidase-encapsulated fluorescent bio-nanoparticle (HEFBNP), comprising bovine serum albumin-stabilized gold nanoclusters (BSA-AuNCs) and horseradish peroxidase-stabilized gold nanoclusters (HRP-AuNCs). The fabricated HEFBNP can sensitively detect H2O2 owing to its two properties. The first is that HEFBNPs have a continuous two-step fluorescence quenching mechanism, which comes from the heterogenous fluorescence quenching mechanism of HRP-AuNCs and BSA-AuNCs. Second, the proximity of two protein-AuNCs in a single HEFBNP allows a reaction intermediate (•OH) to rapidly reach the adjacent protein-AuNCs. As a result, HEFBNP can improve the overall reaction event and decrease the loss of intermediate in the solution. Due to the continuous quenching mechanism and effective reaction event, a HEFBNP-based sensing system can measure very low concentrations of H2O2 up to 0.5 nM and show good selectivity. Furthermore, we design a glass-based microfluidic device to make it easier use HEFBNP, which allowed us to detect H2O2 with the naked eye. Overall, the proposed H2O2 sensing system is expected to be an easy and highly sensitive on-site detection tool in chemistry, biology, clinics, and industry fields.


Asunto(s)
Peróxido de Hidrógeno , Nanopartículas del Metal , Peróxido de Hidrógeno/química , Nanopartículas del Metal/química , Colorantes Fluorescentes/química , Peroxidasa de Rábano Silvestre , Albúmina Sérica Bovina/química , Oro/química , Espectrometría de Fluorescencia
9.
Adv Funct Mater ; 33(46)2023 Nov 09.
Artículo en Inglés | MEDLINE | ID: mdl-39308874

RESUMEN

The human neurovascular system is a complex network of blood vessels and brain cells that is essential to the proper functioning of the brain. In recent years, researchers have become increasingly interested in the role of this system in developing drugs to treat neuroinflammation. This process is believed to contribute to the development of several neurodegenerative diseases, including Alzheimer's and Parkinson's diseases. While much remains to be learned about the precise mechanisms by which the neurovascular system interacts with the brain and how it can be targeted for therapeutic purposes, this area of research holds great promise for the future of neurology and medicine. Currently, creating neurovascular models begins with animal models, followed by testing on humans in clinical trials. However, the high number of medication failures that pass through animal testing indicates that animal models do not always reflect the outcome of human clinical trials. To overcome the challenges of neurovascular systems and the issues with animal models, we have developed a one-of-a-kind in vitro neurovascular unit-on-a-chip to accurately replicate the in vivo human neurovascular microenvironment. This neuroinflammation-on-a-chip platform has the potential to enhance the current methods of drug development and testing to treat neurodegenerative diseases. By replicating the human neurovascular unit in vitro, a more accurate representation of human physiology can be achieved compared to animal models. The ability to detect pro-inflammatory cytokines in situ and monitor physiological changes, such as barrier function, in real-time can provide an invaluable tool for evaluating the efficacy and safety of drugs. Moreover, using nano-sized graphene oxide for in situ detection of inflammatory responses is an innovative approach that can advance the field of neuroinflammation research. Overall, our developed neuroinflammation-on-a-chip system has the potential to provide a more efficient and effective method for developing drugs for treating neurodegenerative diseases and other central nervous system (CNS) diseases.

10.
Biosensors (Basel) ; 12(12)2022 Dec 03.
Artículo en Inglés | MEDLINE | ID: mdl-36551088

RESUMEN

Recently, due to the coronavirus pandemic, the need for early diagnosis of infectious diseases, including viruses, is emerging. Though early diagnosis is essential to prevent infection and progression to severe illness, there are few technologies that accurately measure low concentrations of biomarkers. Plasmonic nanomaterials are attracting materials that can effectively amplify various signals, including fluorescence, Raman, and other optical and electromagnetic output. In this review, we introduce recently developed plasmonic nanobiosensors for measuring viral DNA/RNA as potential biomarkers of viral diseases. In addition, we discuss the future perspective of plasmonic nanobiosensors for DNA/RNA detection. This review is expected to help the early diagnosis and pathological interpretation of viruses and other diseases.


Asunto(s)
Técnicas Biosensibles , Infecciones por Coronavirus , Nanoestructuras , Humanos , ADN Viral , Nanotecnología
11.
Nano Converg ; 9(1): 24, 2022 May 25.
Artículo en Inglés | MEDLINE | ID: mdl-35612632

RESUMEN

There have been several trials to develop the bioactuator using skeletal muscle cells for controllable biobybird robot. However, due to the weak contraction force of muscle cells, the muscle cells could not be used for practical applications such as biorobotic hand for carrying objects, and actuator of biohybrid robot for toxicity test and drug screening. Based on reported hyaluronic acid-modified gold nanoparticles (HA@GNPs)-embedded muscle bundle on PDMS substrate, in this study for augmented actuation, we developed the electroactive nano-biohybrid actuator composed of the HA@GNP-embedded muscle bundle and molybdenum disulfide nanosheet (MoS2 NS)-modified electrode to enhance the motion performance. The MoS2 NS-modified Au-coated polyimide (PI) electrode to be worked in mild pH condition for viable muscle cell was utilized as supporting- and motion enhancing- substrate since it was electrochemically active, which caused the movement of flexible PI electrode. The motion performance of this electroactive nano-biohybrid actuator by electrical stimulation was increased about 3.18 times compared with that of only HA@GNPs embedded-muscle bundle on bare PI substrate. The proposed electroactive nano-biohybrid actuator can be applied to the biorobotic hand and biohybrid robot.

12.
ACS Nano ; 16(4): 5764-5777, 2022 04 26.
Artículo en Inglés | MEDLINE | ID: mdl-35362957

RESUMEN

The detection of nucleic acids and their mutation derivatives is vital for biomedical science and applications. Although many nucleic acid biosensors have been developed, they often require pretreatment processes, such as target amplification and tagging probes to nucleic acids. Moreover, current biosensors typically cannot detect sequence-specific mutations in the targeted nucleic acids. To address the above problems, herein, we developed an electrochemical nanobiosensing system using a phenomenon comprising metal ion intercalation into the targeted mismatched double-stranded nucleic acids and a homogeneous Au nanoporous electrode array (Au NPEA) to obtain (i) sensitive detection of viral RNA without conventional tagging and amplifying processes, (ii) determination of viral mutation occurrence in a simple detection manner, and (iii) multiplexed detection of several RNA targets simultaneously. As a proof-of-concept demonstration, a SARS-CoV-2 viral RNA and its mutation derivative were used in this study. Our developed nanobiosensor exhibited highly sensitive detection of SARS-CoV-2 RNA (∼1 fM detection limit) without tagging and amplifying steps. In addition, a single point mutation of SARS-CoV-2 RNA was detected in a one-step analysis. Furthermore, multiplexed detection of several SARS-CoV-2 RNAs was successfully demonstrated using a single chip with four combinatorial NPEAs generated by a 3D printing technique. Collectively, our developed nanobiosensor provides a promising platform technology capable of detecting various nucleic acids and their mutation derivatives in highly sensitive, simple, and time-effective manners for point-of-care biosensing.


Asunto(s)
Técnicas Biosensibles , COVID-19 , Nanoporos , Ácidos Nucleicos , Humanos , ARN Viral/genética , Técnicas Electroquímicas/métodos , Nucleótidos , SARS-CoV-2 , Electrodos , Técnicas Biosensibles/métodos , Técnicas de Amplificación de Ácido Nucleico/métodos
13.
ACS Sens ; 7(3): 740-747, 2022 03 25.
Artículo en Inglés | MEDLINE | ID: mdl-35138092

RESUMEN

Biohybrid robots, which comprise soft materials with biological components, have the potential to sense, respond, and adapt to changing environmental loads dynamically. Instead of humans and other living things, biohybrid robots can be used in various fields such as drug screening and toxicity assessment. In the actuation part, however, since a muscle cell-based biohybrid robot is limited in that the driving force is weak, it is difficult to evaluate drug and toxicological effects by distinguishing changes in the biohybrid robot's motion. To overcome this limitation, we introduced hyaluronic acid-modified gold nanoparticles (HA-AuNPs) into a muscle bundle-based biohybrid robot that moves forward in response to electrical stimulation. To enhance the actuation of muscle bundles, HA-AuNPs were embedded into the muscle bundles. The motion of the fabricated biohybrid robot was improved due to the enhanced differentiation and the improved electrical conductivity of muscle bundles by HA-AuNPs. In addition, the fabricated biohybrid robot exhibited huge changes in motion with respect to the addition of positive and negative inotropic drugs. The proposed biohybrid robot has the potential for neuromuscular disease drug screening by incorporating nervous tissues such as motor neuron organoids and brain organoids.


Asunto(s)
Nanopartículas del Metal , Robótica , Oro , Humanos , Ácido Hialurónico , Músculo Esquelético/fisiología
14.
Polymers (Basel) ; 14(1)2022 Jan 03.
Artículo en Inglés | MEDLINE | ID: mdl-35012205

RESUMEN

Recently, more and more attention has been paid to the development of eco-friendly solid sorbents that are cost-effective, noncorrosive, have a high gas capacity, and have low renewable energy for CO2 capture. Here, we claimed the fabrication of a three-dimensional (3D) film of hollow nanocones with a large surface area (949.5 m2/g), a large contact angle of 136.3°, and high surface energy. The synthetic technique is based on an electrochemical polymerization process followed by a novel and simple strategy for pulling off the formed layers as a membrane. Although the polymer-coated substrates were reported previously, the membrane formation has not been reported elsewhere. The detachable capability of the manufactured layer as a membrane braked the previous boundaries and allows the membrane's uses in a wide range of applications. This 3D hollow nanocones membrane offer advantages over conventional ones in that they combine a π-electron-rich (aromatic ring), hydrophobicity, a large surface area, multiple amino groups, and a large pore volume. These substantial features are vital for CO2 capturing and storage. Furthermore, the hydrophobicity characteristic and application of the formed polymer as a CO2 sucker were investigated. These results demonstrated the potential of the synthesized 3D hollow polymer to be used for CO2 capturing with a gas capacity of about 68 mg/g and regeneration ability without the need for heat up.

15.
J Control Release ; 342: 228-240, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-35016917

RESUMEN

RNA interference (RNAi) is being extensively investigated as a potential therapeutic strategy for cancer treatment. However, RNAi-based therapeutics have not yet been used to treat cancer because of their instability and the difficulty of microRNA (miRNA) delivery. Plasmonic nanoparticle-based RNAi nanotherapeutics have been developed for accurate and sensitive diagnosis and a strong therapeutic effect on cancers by leveraging their ease-of-use and specific properties such as photothermal conversion. In this review, recent strategies and advances in plasmonic nanoparticle-based miRNA delivery are briefly presented to facilitate the detection and treatment of several cancers. The challenges and potential opportunities afforded by the RNAi-based theragnosis field are discussed. We expect that the RNAi-integrated plasmonic nanotherapeutics discussed in this review can provide insights for the early diagnosis and effective treatment of cancer.


Asunto(s)
MicroARNs , Nanopartículas , Neoplasias , Sistemas de Liberación de Medicamentos , Humanos , MicroARNs/genética , MicroARNs/uso terapéutico , Neoplasias/diagnóstico , Neoplasias/genética , Neoplasias/terapia , Interferencia de ARN , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/uso terapéutico
16.
Anal Bioanal Chem ; 414(10): 3219-3230, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34767053

RESUMEN

Microfluidic-based biosensors have been developed for their precise automatic reaction control. However, these biosensors require external devices that are difficult to transport and use. To overcome this disadvantage, our group made an easy-to-use, cheap, and light pumpless three-dimensional photo paper-based microfluidic analytical device (3D-µPAD; weight: 1.5 g). Unlike conventional paper-based microfluidic analytical devices, the 3D-µPAD can be used to control fluid flow in a 3D manner, thus allowing sophisticated multi-step reaction control. This device can control fluid flow speed and direction accurately using only the capillary-driven flow without an external device like a pump. The flow speed is controlled by the width of the microfluidic channel and its surface property. In addition, fluid speed control and 3D-bridge structure enable the control of fluid flow direction. Using these methods, multi-step enzyme-linked immunosorbent assay (ELISA) can be done automatically in sequence by injecting solutions (sample, washing, and enzyme's substrate) at the same time in the 3D-µPAD. All the steps can be performed in 14 min, and data can be analyzed immediately. To test this device, thioredoxin-1 (Trx-1), a biomarker of breast cancer, is used as the target. In the 3D-µPAD, it can detect 0-200 ng/mL of Trx-1, and the prepared 3D-µPAD Trx-1 sensor displays excellent selectivity. Moreover, by analyzing the concentration of Trx-1 in real patients and healthy individuals' blood serum samples using the 3D-µPAD, and comparing results to ELISA, it can be confirmed that the 3D-µPAD is a good tool for cancer diagnosis.


Asunto(s)
Técnicas Analíticas Microfluídicas , Microfluídica , Ensayo de Inmunoadsorción Enzimática , Humanos , Papel , Tiorredoxinas
17.
Biomedicines ; 9(8)2021 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-34440132

RESUMEN

Nucleic acids, including DNA and RNA, have received prodigious attention as potential biomarkers for precise and early diagnosis of cancers. However, due to their small quantity and instability in body fluids, precise and sensitive detection is highly important. Taking advantage of the ease-to-functionality and plasmonic effect of nanomaterials, fluorescence resonance energy transfer (FRET) and metal-enhanced fluorescence (MEF)-based biosensors have been developed for accurate and sensitive quantitation of cancer-related nucleic acids. This review summarizes the recent strategies and advances in recently developed nanomaterial-based FRET and MEF for biosensors for the detection of nucleic acids in cancer diagnosis. Challenges and opportunities in this field are also discussed. We anticipate that the FRET and MEF-based biosensors discussed in this review will provide valuable information for the sensitive detection of nucleic acids and early diagnosis of cancers.

18.
ACS Nano ; 15(8): 13475-13485, 2021 08 24.
Artículo en Inglés | MEDLINE | ID: mdl-34369760

RESUMEN

Nucleic acid biomarkers have been widely used to detect various viral-associated diseases, including the recent pandemic COVID-19. The CRISPR-Cas-based trans-activating phenomenon has shown excellent potential for developing sensitive and selective detection of nucleic acids. However, the nucleic acid amplification steps are typically required when sensitive and selective monitoring of the target nucleic acid is needed. To overcome the aforementioned challenges, we developed a CRISPR-Cas12a-based nucleic acid amplification-free biosensor by a surface-enhanced Raman spectroscopy (SERS)-assisted ultrasensitive detection system. We integrated the activated CRISPR-Cas12a by viral DNA with a Raman-sensitive system composed of ssDNA-immobilized Raman probe-functionalized Au nanoparticles (RAuNPs) on the graphene oxide (GO)/triangle Au nanoflower array. Using this CRISPR-based Raman-sensitive system improved the detection sensitivity of the multiviral DNAs such as hepatitis B virus (HBV), human papillomavirus 16 (HPV-16), and HPV-18 with an extremely low detection limit and vast detection range from 1 aM to 100 pM without the amplification steps. We suggest that this ultrasensitive amplification-free detection system for nucleic acids can be widely applied to the precise and early diagnosis of viral infections, cancers, and several genetic diseases.


Asunto(s)
Técnicas Biosensibles , COVID-19 , Nanopartículas del Metal , Ácidos Nucleicos , Humanos , Espectrometría Raman/métodos , ADN Viral/genética , Oro/química , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas Biosensibles/métodos
19.
Cancers (Basel) ; 13(6)2021 Mar 18.
Artículo en Inglés | MEDLINE | ID: mdl-33803846

RESUMEN

Detecting circulating tumor cells (CTCs) has been considered one of the best biomarkers in liquid biopsy for early diagnosis and prognosis monitoring in cancer. A major challenge of using CTCs is detecting extremely low-concentrated targets in the presence of high noise factors such as serum and hematopoietic cells. This review provides a selective overview of the recent progress in the design of microfluidic devices with optical sensing tools and their application in the detection and analysis of CTCs and their small malignant subset, circulating cancer stem cells (CCSCs). Moreover, discussion of novel strategies to analyze the differentiation of circulating cancer stem cells will contribute to an understanding of metastatic cancer, which can help clinicians to make a better assessment. We believe that the topic discussed in this review can provide brief guideline for the development of microfluidic-based optical biosensors in cancer prognosis monitoring and clinical applications.

20.
Nano Lett ; 21(1): 693-699, 2021 01 13.
Artículo en Inglés | MEDLINE | ID: mdl-33346665

RESUMEN

Cell-free DNA (cfDNA) has attracted significant attention due to its high potential to diagnose diseases, such as cancer. Still, its detection by amplification method has limitations because of false-positive signals and difficulty in designing target-specific primers. CRISPR-Cas-based fluorescent biosensors have been developed but also need the amplification step for the detection. In this study, for the first time CRISPR-Cas12a based nucleic acid amplification-free fluorescent biosensor was developed to detect cfDNA by a metal-enhanced fluorescence (MEF) using DNA-functionalized Au nanoparticle (AuNP). Upon activating the CRISPR-Cas12a complex by the target cfDNA and subsequent single-strand DNA (ssDNA) degradation between AuNP and fluorophore, MEF occurred with color changes from purple to red-purple. Using this system, breast cancer gene-1 (BRCA-1) can be detected with very high sensitivity in 30 min. This rapid and highly selective sensor can be applied to measure other nucleic acid biomarkers such as viral DNA in field-deployable and point-of-care testing (POCT) platform.


Asunto(s)
Técnicas Biosensibles , Nanopartículas del Metal , Ácidos Nucleicos , Sistemas CRISPR-Cas/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Colorimetría , ADN/genética , Oro
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