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1.
Mikrochim Acta ; 191(10): 594, 2024 09 12.
Article in English | MEDLINE | ID: mdl-39264373

ABSTRACT

A sandwich electrochemical sensor was fabricated based on multi-walled carbon nanotubes/ordered mesoporous carbon/AuNP (MWCNTs/CMK-3/AuNP) nanocomposites and porous core-shell nanoparticles Au@PdNPs to achieve rapid and sensitive detection of AFB1 in complex matrices. MWCNTs/CMK-3/AuNP nanocomposite, which was prepared by self-assembly method, served as a substrate material to increase the aptamer loading and improve the conductivity and electrocatalytic activity of the electrode for the first signal amplification. Then, Au@PdNPs, which were synthesized by one-pot aqueous phase method, were applied as nanocarriers loaded with plenty of capture probe antibody (Ab) and signal molecule toluidine blue (Tb) to form the Au@PdNPs-Ab-Tb bioconjugates for secondary signal amplification. The sensing system could still significantly improve the signal output intensity even in the presence of ultra-low concentration target compound due to the dual signal amplification of MWCNTs/CMK-3/AuNP nanocomposites and Au@PdNPs-Ab-Tb. The method exhibited high selectivity, low detection limit (9.13 fg/mL), and strong stability to differentiate AFB1 from other mycotoxins. Furthermore, the sensor has been successfully applied to the quantitative determination of AFB1 in corn, malt, and six herbs, which has potential applications in food safety, quality control, and environmental monitoring.


Subject(s)
Aflatoxin B1 , Electrochemical Techniques , Gold , Limit of Detection , Metal Nanoparticles , Nanotubes, Carbon , Palladium , Gold/chemistry , Metal Nanoparticles/chemistry , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Palladium/chemistry , Aflatoxin B1/analysis , Aflatoxin B1/immunology , Nanotubes, Carbon/chemistry , Biosensing Techniques/methods , Antibodies, Immobilized/immunology , Nanocomposites/chemistry , Aptamers, Nucleotide/chemistry , Food Contamination/analysis , Zea mays/chemistry , Electrodes
2.
Bioelectrochemistry ; 160: 108793, 2024 Dec.
Article in English | MEDLINE | ID: mdl-39128408

ABSTRACT

Estrogen receptor alpha (ERα) serves as a crucial biomarker for early breast cancer diagnosis. In this study, we proposed an electrochemical aptasensor with nanomaterial carbon nanohorns/gold nanoparticle composites (1-AP-CNHs/AuNPs) as the substrate, and the primary amine groups on the antibody initiated the ring-opening polymerization (ROP) of monomer amino acid-ferrocene (NCA-Fc) on the electrode surface for ultrasensitive detection of ERα. The composite of 1-AP-CNHs/AuNPs not only possessed more active sites, but also increased the specific surface area of the electrode and allowed a large amount of ferrocene polymer long chains to be grafted onto the electrode surface to achieve signal amplification. Under optimal conditions, the detection limit of the method was 11.995 fg mL-1 with a detection range of 100 fg mL-1-100 ng mL-1. In addition, the biotin-streptavidin system was used to further improve the sensitivity of the sensor. Importantly, this approach could be applied for the practical detection of ERα in real samples.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Electrochemical Techniques , Estrogen Receptor alpha , Gold , Limit of Detection , Metal Nanoparticles , Gold/chemistry , Electrochemical Techniques/methods , Humans , Metal Nanoparticles/chemistry , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Polymerization , Metallocenes/chemistry , Ferrous Compounds/chemistry , Electrodes
3.
Talanta ; 278: 126468, 2024 Oct 01.
Article in English | MEDLINE | ID: mdl-38963975

ABSTRACT

Carcinoembryonic Antigen (CEA), an acidic glycoprotein with human embryonic antigen properties, is found on the surface of cancer cells that have differentiated from endodermal cells. This paper presents a label-free electrochemical immunoassay for the dual amplification detection of CEA using gold nanoparticles loaded with polypyrrole polydopamine (Au/PPy-PDA) and polymerized polycaprolactone (Ng-PCL) prepared by ring-opening polymerization (ROP). First, the composite Au/PPy-PDA was adhered to the electrode surface. Then, gold nanoparticles form a Au-S bond with the sulfhydryl group in Apt1 to secure it on the electrode surface. Subsequently, the non-specific binding sites on the electrodes surface are closed by bovine serum albumin (BSA). Next, CEA is dropped onto the electrode surface, which is immobilized by antigen-antibody specific recognition, and the carboxyl-functionalized Apt2 forms a "sandwich structure" of antibody-antigen-antibody by specific recognition. Polymeric Ng-PCL is adhered to the electrode surface, leading to an increase in the electrochemical impedance signal, resulting in a complete chain of signal analysis. Finally, the response signal is detected by electrochemical impedance spectroscopy (EIS). Under optimal experimental conditions, the method has the advantages of high sensitivity and wide linear range (1 pg mL-1∼100 ng mL-1), and the lower limit of detection (LOD) is 0.234 pg mL-1. And it has the same high sensitivity, selectivity and interference resistance for the real samples detection. Thus, it provides a new way of thinking about biomedical and clinical diagnosis.


Subject(s)
Biosensing Techniques , Carcinoembryonic Antigen , Electrochemical Techniques , Gold , Metal Nanoparticles , Polyesters , Polymers , Gold/chemistry , Biosensing Techniques/methods , Metal Nanoparticles/chemistry , Carcinoembryonic Antigen/analysis , Carcinoembryonic Antigen/immunology , Polyesters/chemistry , Electrochemical Techniques/methods , Polymers/chemistry , Humans , Indoles/chemistry , Immunoassay/methods , Limit of Detection , Electrodes , Pyrroles/chemistry , Serum Albumin, Bovine/chemistry
4.
Luminescence ; 39(6): e4804, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38859763

ABSTRACT

Early and sensitive detection of tobacco mosaic virus (TMV) is of great significance for improving crop yield and protecting germplasm resources. Herein, we constructed a novel fluorescence sensor to detect TMV RNA (tRNA) through double strand specific nuclease (DSN) cycle and activator regenerative electron transfer atom transfer radical polymerization (ARGET ATRP) dual signal amplification strategy. The hairpin DNA complementarily paired with tRNA was used as a recognition unit to specifically capture tRNA. By the double-stranded DNA hydrolyzed with DSN, tRNA is released to open more hairpin DNA, and more complementary DNA (cDNA) is bound to the surface of the magnetic beads (MBs) to achieve the first amplification. After binding with the initiator, the cDNA employed ARGET ATRP to attach more fluorescent signal molecules to the surface of MBs, thus achieving the second signal amplification. Under the optimal experimental conditions, the logarithm of fluorescence intensity versus tRNA concentration showed a good linear relationship in the range of 0.01-100 pM, with a detection limit of 1.03 fM. The limit of detection (LOD) was calculated according to LOD = 3 N/S. Besides, the sensor showed good reproducibility and stability, which present provided new method for early and highly sensitive detection for plant viruses.


Subject(s)
RNA, Viral , Tobacco Mosaic Virus , Tobacco Mosaic Virus/genetics , Tobacco Mosaic Virus/chemistry , RNA, Viral/analysis , Fluorescence , Limit of Detection , Biosensing Techniques/methods , Fluorescent Dyes/chemistry , Spectrometry, Fluorescence
5.
J Mater Chem B ; 12(24): 5861-5868, 2024 Jun 19.
Article in English | MEDLINE | ID: mdl-38775046

ABSTRACT

The development of a simple, rapid, and sensitive technology for the simultaneous detection of mycotoxins is of great significance in ensuring the safety of foods and drugs. Herein, a fluorescence aptasensor with high sensitivity and reproducibility for the simultaneous detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA) was developed. In this sensing system, AFB1 and OTA aptamers were co-immobilized on the surface of magnetic beads (MBs) to form a Y-shaped structure through the principle of complementary base pairing, and were used as recognition probes to specifically capture the target. Activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP) was used as a signal amplification strategy to improve the sensitivity. The initiator modified at the end of an antibody initiates the ARGET ATRP reaction. Different fluorescence signals were designed to achieve the simultaneous detection of OTA and AFB1 with limits of 426.18 and 79.55 fg mL-1 for AFB1 and OTA, respectively. In addition, experiments were conducted on three types of samples, and the recoveries of the two mycotoxins ranged from 87.30% to 109.50%, with relative standard deviations ranging from 0.50% to 4.92% under reproducible conditions. The results suggest that the developed aptasensor is sufficient to meet the different regulatory requirements of the two mycotoxins in food and drug safety and shows great potential.


Subject(s)
Aflatoxin B1 , Aptamers, Nucleotide , Biosensing Techniques , Ochratoxins , Aflatoxin B1/analysis , Ochratoxins/analysis , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , DNA/chemistry , Polymerization , Limit of Detection , Electron Transport
6.
Mikrochim Acta ; 191(6): 348, 2024 05 28.
Article in English | MEDLINE | ID: mdl-38805077

ABSTRACT

A novel aptamer-based sensor was developed using the signal amplification strategy of ring-opening metathesis polymerization (ROMP) and polyethyleneimine modified graphene oxide to achieve trace detection of carbendazim (CBZ). The dual identification of aptamer and antibody was used to avoid false positive results and improve the selectivity. Polyethyleneimine modified graphene oxide (GO-PEI), as a substrate material with excellent conductivity, was modified on the surface of a glassy carbon electrode (GCE) to increase the grafting amount of aptamer on the electrode surface. Moreover, a large number of cyclopentenyl ferrocene (CFc) was aggregated to form long polymer chains through ring-opening metathesis polymerization (ROMP), so as to significantly improve the detection sensitivity of the biosensor. The linear range of this sensor was 1 pg/mL-100 ng/mL with a detection limit as low as 7.80 fg/mL. The sensor exhibited excellent reproducibility and stability, and also achieved satisfactory results in actual sample detection. The design principle of such a sensor could provide innovative ideas for sensors in the detection of other types of targets.


Subject(s)
Aptamers, Nucleotide , Benzimidazoles , Biosensing Techniques , Carbamates , Electrochemical Techniques , Graphite , Limit of Detection , Polyethyleneimine , Polymerization , Graphite/chemistry , Carbamates/chemistry , Carbamates/analysis , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Polyethyleneimine/chemistry , Biosensing Techniques/methods , Benzimidazoles/chemistry , Aptamers, Nucleotide/chemistry , Electrodes , Reproducibility of Results
7.
Chempluschem ; 89(8): e202400119, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38619207

ABSTRACT

Down-regulator of transcription 1 (DR1) is considered as a biomarker of hashimoto's thyroiditis (HT), which is a risk factor for thyroid cancer. Here, a label-free electrochemical biosensor for DR1 detection was constructed based on polyamidoamine (PAMAM) polymer and the nanocomposite (WO3@AuNPs) composed of tungsten trioxide (WO3) and gold nanoparticles (AuNPs). WO3@AuNPs was obtained by combining monolayer WO3 nanosheets, which has high conductivity, and AuNPs. The modification of WO3@AuNPs can not only increase the conductivity of the electrode but also provide more active sites for signaling units, thus greatly improve the sensitivity of the sensor. The polymer PAMAM is biocompatible and non-immunogenic, and its end functional group can bind to the target molecules, providing them with more binding sites and thus improving the sensitivity of the sensor. Under optimal conditions, the label-free biosensor showed a good linear relationship between the logarithm of DR1 concentration and the impedance in the range of 10 fg ⋅ mL-1 to 100 ng ⋅ mL-1, with a detection limit as low as 0.3 fg ⋅ mL-1. Besides, this label-free electrochemical platform exhibited satisfactory selectivity and anti-interference capability in human serum samples. Therefore, this method has considerable potential in clinical detection of DR1.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Gold , Metal Nanoparticles , Oxides , Tungsten , Gold/chemistry , Metal Nanoparticles/chemistry , Electrochemical Techniques/methods , Tungsten/chemistry , Humans , Oxides/chemistry , Biosensing Techniques/methods , Polyamines/chemistry , Limit of Detection
8.
Noise Health ; 26(120): 19-24, 2024.
Article in English | MEDLINE | ID: mdl-38570306

ABSTRACT

BACKGROUND: Patients undergoing total knee arthroplasty (TKA) need to tolerate the effects of noise. MATERIALS AND METHODS: This study retrospectively analyzed the clinical data of 167 TKA patients at The Affiliated Hospital of Southwest Medical University from April 2019 to April 2021. A total of 154 patients who met inclusion criteria were divided into the conventional noise reduction management group (CMG) and the noise reduction earplug group (EPG), following different management schemes. The CMG received routine noise reduction management after surgery, while the EPG used noise reduction earplugs based on the CMG. The clinical indexes of the two groups were compared. RESULTS: In this study, 79 patients were included in the CMG, and 75 patients were included in the EPG. The results showed that the Pittsburgh Sleep Quality Index (PSQI) scores of both groups 2 weeks after surgery were significantly lower than those before management (ZEPG = 5.995, ZCMG = 4.109, all P < 0.001), and the EPG exhibited a significantly lower PSQI score than the CMG (Z = -2.442, P < 0.05). Two weeks after surgery, the EPG had significantly lower levels of systolic blood pressure (ZSBP = -4.303) and diastolic blood pressure (ZDBP = -3.115), as well as lower scores on the Hospital Anxiety and Depression Scale-Anxiety (HADS-A; ZHADS-A = -7.140) and Hospital Anxiety and Depression Scale-Depression (HADS-D; ZHADS-D = -4.545) compared to the CMG (all P < 0.05). In addition, no significant correlation existed between the duration of wearing earplugs and the HADS-A and HADS-D scores (r = -0.201, r = -0.002, P > 0.05). CONCLUSION: Noise reduction earplugs can improve sleep quality and regulate negative emotions of patients undergoing TKA treatment through a complex mechanism involving noise, which is beneficial to the prognosis of the disease.


Subject(s)
Arthroplasty, Replacement, Knee , Humans , Retrospective Studies , Ear Protective Devices , Noise/adverse effects
9.
Talanta ; 275: 126130, 2024 Aug 01.
Article in English | MEDLINE | ID: mdl-38653117

ABSTRACT

Human epidermal growth factor receptor 2 (HER2), a common proto-oncogene, is overexpressed in a subset of breast cancer patients. It is essential to track HER2 expression for early breast cancer diagnosis. Herein, a ratiometric electrochemical biosensor for detection of HER2 based on activators generated by electron transfer for atom transfer radical polymerisation (AGET ATRP) and hairpin DNA was developed. Specifically, hairpin DNA was first self-assembled on the gold electrode by Au-S bond. Upon capturing HER2, the stem-loop structure of hairpin DNA was unfolded, the signal value of methylene blue (MB) decreased as it moved away from the electrode surface. cDNA was linked with HER2 by complementary base pairing to introduce amino group. Then, the initiator 2-bromo-2-methylpropionic acid (BMP) were connected to the amino group on the cDNA to activate ARGET ATRP. The detection performance of biosensors for HER2 was explored by the ratio signal between two signal molecules. Under optimal conditions, this ratiometric electrochemical biosensor shows good selectivity and stability with a wide detection range of 1-1 × 106 pM and a detection limit of 78.47 fM. Furthermore, the biosensor exhibits satisfactory anti-interference ability due to the hairpin DNA and dual signal system, and has promising application prospects in the detection of other DNA disease markers.


Subject(s)
Biosensing Techniques , Electrochemical Techniques , Proto-Oncogene Mas , Receptor, ErbB-2 , Biosensing Techniques/methods , Receptor, ErbB-2/genetics , Humans , Electrochemical Techniques/methods , Electrodes , Gold/chemistry , Limit of Detection , Polymerization , DNA/chemistry , DNA/genetics
10.
Mikrochim Acta ; 191(3): 148, 2024 02 20.
Article in English | MEDLINE | ID: mdl-38374311

ABSTRACT

A unique combination of a specific nucleic acid restriction endonuclease (REase) and atom transfer radical polymerization (ATRP) signal amplification strategy was employed for the detection of T790M mutations prevalent in the adjuvant diagnosis of lung cancer. REase selectively recognizes and cleaves T790M mutation sites on double-stranded DNA formed by hybridization of a capture sequence and a target sequence. At the same time, the ATRP strategy resulted in the massive aggregation of upconverted nanoparticles (UCNPs), which significantly improved the sensitivity of the biosensor. In addition, the UCNPs have excellent optical properties and can eliminate the interference of autofluorescence in the samples, thus further improving the detection sensitivity. The proposed upconversion fluorescent biosensor is characterized by high specificity, high sensitivity, mild reaction conditions, fast response time, and a detection limit as low as 0.14 fM. The performance of the proposed biosensor is comparable to that of clinical PCR methods when applied to clinical samples. This work presents a new perspective for assisted diagnosis in the pre-intervention stage of tumor diagnostics in the early stage of precision oncology treatments.


Subject(s)
Biosensing Techniques , Lung Neoplasms , Humans , Lung Neoplasms/genetics , DNA Restriction Enzymes , ErbB Receptors/genetics , Polymerization , DNA Cleavage , Limit of Detection , Mutation , Precision Medicine , Protein Kinase Inhibitors , Biosensing Techniques/methods
11.
Mikrochim Acta ; 190(11): 432, 2023 10 09.
Article in English | MEDLINE | ID: mdl-37806989

ABSTRACT

An ochratoxin A (OTA) electrochemical biosensor based on a cascade signal amplification strategy with Ag nanoparticles (AgNPs) and ring opening polymerization (ROP) was constructed. The large specific surface area of AgNPs was used to increase the loading of OTA aptamer on the electrode surface, enhancing the ability to capture OTA as a way to achieve the first signal amplification. The OTA antibody modified with polyethylenimine specifically recognizes the OTA, forming an aptamer-OTA-antibody sandwich structure. The amino group on polyethylenimine initiates the ROP reaction with α-amino acid-n-carboxylic anhydride-ferrocene (NCA-Fc) as the monomer. A large number of electrochemical signal units of ferrocene are introduced into the sensing system for a second signal amplification. By amplifying the signal twice, the sensitivity of the sensor is improved. Under the optimal conditions, the detection range of the sensor is 1 pg·mL-1 ~ 1 µg·mL-1, while the detection limit is as low as 117 fg·mL-1. Moreover, the sensor has the advantages of high sensitivity, good stability and selectivity. Standard addition recovery experiment proved that the sensing system can be successfully used for the detection of OTA in four actual samples with recoveries in the range 90.0 to 113% with RSDs of 0.6 to 5.2%, providing a new idea for the pollution assessment of mycotoxins.


Subject(s)
Biosensing Techniques , Metal Nanoparticles , Metallocenes/chemistry , Metal Nanoparticles/chemistry , Polyethyleneimine , Polymerization , Electrochemical Techniques , Silver
12.
Chemistry ; 29(65): e202301602, 2023 Nov 21.
Article in English | MEDLINE | ID: mdl-37622405

ABSTRACT

The levels of KRAS G12C point mutation is recognized to be closely related to the earlier diagnosis of non-small cell lung cancer (NSCLC). Here, based on nitrogen-doped graphene quantum dots (NGQDs) and photo-induced electron/energy transfer reversible addition-fragment chain transfer (PET-RAFT) signal amplification strategy, we fabricated a novel electrochemiluminescence (ECL) biosensor for the detection of KRAS G12C mutation for the first time. NGQDs as ECL-emitting species with cathodic ECL were prepared by a simple calcination method. Firstly, KRAS G12C mutation DNA, i. e., target DNA (tDNA), was captured by specific identification with hairpin DNA (hDNA). Then, PET-RAFT was initiated by blue light, and large numbers of monomers were successfully polymerized to form controllable polymer chains. Lastly, massive NGQDs was introduced via amidation reaction with N-(3-aminopropyl)methacrylamide hydrochloride (APMA), which significantly amplified the ECL signal intensity. Under optimal conditions, this biosensor achieved a good linear relationship between ECL intensity and logarithm of the levels of KRAS G12C mutation in the range from 10 fM to 10 nM. Moreover, this strategy exhibited high selectivity and excellent applicability for KRAS G12C mutation detection in the serum samples. Therefore, this biosensor has great potential in clinical diagnosis and practical application.


Subject(s)
Biosensing Techniques , Carcinoma, Non-Small-Cell Lung , Graphite , Lung Neoplasms , Quantum Dots , Humans , Proto-Oncogene Proteins p21(ras)/genetics , Nitrogen , Luminescent Measurements/methods , DNA , Biosensing Techniques/methods , Mutation , Positron-Emission Tomography
13.
Talanta ; 262: 124659, 2023 Sep 01.
Article in English | MEDLINE | ID: mdl-37220688

ABSTRACT

Accurate and ultrasensitive detection of cytokeratin 19 fragment (CYFRA21-1) is of vital importance for screening and diagnosis of potential lung cancer patient. In this paper, surface-modified upconversion nanomaterials (UCNPs) capable of aggregation by atom transfer radical polymerization (ATRP) were used as luminescent materials for the first time to achieve signal-stable, low-biological background, and sensitive detection of CYFRA21-1. Upconversion nanomaterials (UCNPs) feature extremely low biological background signals and narrow emission peaks, making them ideal sensor luminescent materials. The combination of UCNPs and ATRP not only improves sensitivity, but also reduces biological background interference for detecting CYFRA21-1. The target CYFRA21-1 was captured by specific binding of the antigen and the antibody. Subsequently, the end of the sandwich structure with the initiator reacts with monomers modified on UCNPs. Then, massive UCNPs are aggregated by ATRP that amplify the detection signal exponentially. Under optimal conditions, a linear calibration plot of the logarithm of CYFRA21-1 concentration versus the upconversion fluorescence intensity was obtained in the range of 1 pg/mL to 100 µg/mL with a detection limit of 38.7 fg/mL. The proposed upconversion fluorescent platform can distinguish the analogues of the target with excellent selectivity. Besides, the precision and accuracy of the developed upconversion fluorescent platform were verified by clinical methods. As an enhanced upconversion fluorescent platform of CYFRA21-1, it is expected to be useful in screening potential patients with NSCLC and provides a promising solution for the high-performance detection of other tumor markers.


Subject(s)
Biosensing Techniques , Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Nanoparticles , Humans , Antigens, Neoplasm , Keratin-19 , Lung Neoplasms/diagnosis , Biosensing Techniques/methods , Limit of Detection , Nanoparticles/chemistry
14.
Food Chem ; 421: 136176, 2023 Sep 30.
Article in English | MEDLINE | ID: mdl-37098309

ABSTRACT

An electrochemical sensor based on environmentally friendly eRAFT polymerization was developed for the detection of aflatoxin B1 (AFB1) in food and herbal medicine. Two biological probes, aptamer (Ap) and antibody (Ab), were used to specifically recognize AFB1, and a large number of ferrocene polymers were grafted on the electrode surface by eRAFT polymerization, which greatly improved the specificity and sensitivity of the sensor. The detection limit of AFB1 was 37.34 fg/mL. In addition, the recovery rate was 95.69% to 107.65% and the RSD was 0.84% to 4.92% by detecting 9 spiked samples. The delighted reliability of this method was verified by HPLC-FL.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Polymers , Reproducibility of Results , Biosensing Techniques/methods , Electrochemical Techniques/methods , Aflatoxin B1/analysis , Limit of Detection
15.
Bioelectrochemistry ; 151: 108402, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36841148

ABSTRACT

Cardiac troponin I (cTnI) is considered as the gold standard for the diagnosis of acute myocardial infarction (AMI) because of its excellent specificity and sensitivity. Herein, a novel aptasensor based on the dual signal amplification strategy of Polyethyleneimine functionalized Graphene oxide (GO) and ring-opening polymerization (ROP) for the first time was successfully constructed to achieve high sensitivity detection of cTnI. Briefly, cTnI-aptamer 1 (Apt1) was immobilized on the surface of gold electrode by self-assembly of Au-S bonds to specifically capture cTnI. After specific recognition of cTnI, Apt2 coated PEI-functionalized GO composites acted as macroinitiators for the subsequent ROP reaction. Next, α-amino acid-N-carboxylic acid anhydride ferrocene derivatives (NCA-Fc), the monomer for ROP reaction, was added to the electrode surface. The combined application of PEI-functionalized GO and NCA-Fc better achieves the high sensitivity and signal amplification of the aptasensor. Under optimal conditions, the aptasensor exhibited a wide linear range of 10 fg mL-1 to 10 ng mL-1 and the limit of detection was 3.78 fg mL-1. Moreover, this method displayed the advantages of good selectivity, simple operation and excellent stability. Meanwhile, the aptasensor had good accuracy and applicability even in real serum samples analysis, demonstrating its considerable application potential in biomedical assays.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Graphite , Metal Nanoparticles , Limit of Detection , Troponin I/analysis , Polymerization , Aptamers, Nucleotide/chemistry , Graphite/chemistry , Electrochemical Techniques/methods , Biosensing Techniques/methods , Metal Nanoparticles/chemistry
16.
Talanta ; 257: 124360, 2023 May 15.
Article in English | MEDLINE | ID: mdl-36801566

ABSTRACT

Plant diseases caused by tobacco mosaic viruses (TMV) reduce the yield and quality of crops and cause significant losses. Early detection and prevention of TMV has important value of research and reality. Herein, a fluorescent biosensor was constructed for highly sensitive detection of TMV RNA (tRNA) based on the principle of base complementary pairing, polysaccharides and atom transfer radical polymerization by electron transfer activated regeneration catalysts (ARGET ATRP) as double signal amplification strategy. The 5'-end sulfhydrylated hairpin capture probe (hDNA) was first immobilized on amino magnetic beads (MBs) by a cross-linking agent, which specifically recognizes tRNA. Then, chitosan binds to BIBB, providing numerous active sites for fluorescent monomer polymerization, which successfully significantly amplifying the fluorescent signal. Under optimal experimental conditions, the proposed fluorescent biosensor for the detection of tRNA has a wide detection range from 0.1 pM to 10 nM (R2 = 0.998) with a limit of detection (LOD) as low as 1.14 fM. In addition, the fluorescent biosensor showed satisfactory applicability for the qualitative and quantitative analysis of tRNA in real samples, thereby demonstrating the potential in the field of viral RNA detection.


Subject(s)
Biosensing Techniques , Tobacco Mosaic Virus , RNA , Polysaccharides , Limit of Detection
17.
Anal Biochem ; 660: 114971, 2023 01 01.
Article in English | MEDLINE | ID: mdl-36328214

ABSTRACT

Exosome is an emerging tumor marker, whose concentration level can reflect the occurrence and development of tumors. The development of rapid and sensitive exosome detection platform is of great significance for early warning of cancer occurrence. Here, a strategy for electrochemical detection of A549-cell-derived exosomes was established based on DNA/ferrocene-modified single-walled carbon nanotube complex (DNA/SWCNT-Fc). DNA/SWCNT-Fc complexes function as a signal amplification platform to promote electron transfer between electrochemical signal molecules and electrodes, thereby improving sensitivity. At the same time, the exosomes can be attached to DNA/SWCNT-Fc nanocomposites via the established PO43--Ti4+-PO43- method. Moreover, the application of EGFR antibody, which can specifically capture A549 exosomes, could improve the accuracy of this sensing system. Under optimal experimental conditions, the biosensor showed good linear relationship between the peak current and the logarithm of exosomes concentration from 4.66 × 106 to 9.32 × 109 exosomes/mL with a detection limit of 9.38 × 104 exosomes/mL. Furthermore, this strategy provides high selectivity for exosomes of different cancer cells, which can be applied to the detection of exosomes in serum samples. Thus, owing to its advantages of high sensitivity and good selectivity, this method provides a diversified platform for exosomes identification and has great potential in early diagnosis and biomedical applications.


Subject(s)
Exosomes , Nanotubes, Carbon , Metallocenes , DNA
18.
Talanta ; 252: 123775, 2023 Jan 15.
Article in English | MEDLINE | ID: mdl-36037766

ABSTRACT

Alkaline phosphatase (ALP) is a significant hydrolase enzyme found in living organisms, and the dysregulation of its physiological activity has been correlated with a variety of diseases. Exploring the activity of ALP has important implications for biomedical research and clinical diagnosis. Accordingly, we have developed a novel, highly sensitive electrochemical biosensor for the analysis of ALP. Based on photoinduced atom transfer radical polymerisation (photoATRP), this strategy combined a fabricated biosensor with hydrolysate produced by the hydrolysis of O-phosphoethanolamine by ALP. Furthermore, for signal amplification, photoATRP synthesises uses polymers with plentiful binding sites for ferrocenylmethyl methacrylate, and by using a photoredox catalyst under blue light irradiation to perform this without the need for copper complexes, it is beneficial for environmental protection compared to traditional atom transfer radical polymerisation (ATRP). The biosensor had a linear range of 10-150 mU·mL-1, with R2 = 0.998, and detection limits as low as 2.12 mU·mL-1. Moreover, by exhibiting outstanding selectivity and interference resistance in human serum samples, this sensor has great potential for practical applications.


Subject(s)
Alkaline Phosphatase , Biosensing Techniques , Humans , Polymerization , Biological Assay , Catalysis , Limit of Detection , Electrochemical Techniques
19.
Anal Biochem ; 655: 114834, 2022 10 15.
Article in English | MEDLINE | ID: mdl-35940299

ABSTRACT

Herein, an electroluminescence (ECL) biosensor was constructed by combining click chemistry with activators regenerated by electron transfer-atom transfer radical polymerization (ARGET-ATRP) to sensitively assay tobacco mosaic virus (TMV) RNA for the first time. First, hairpin DNA (hDNA) was self-assembled on the gold electrode surface through Au-S bonding. The hDNA hybridized with the tDNA to form tRNA/hDNA hybrids in the presence of TMV RNA (tRNA), so that the azide group labelled at the end of the hDNA was kept away from the electrode surface. Subsequently, the initiator for the ARGET-ATRP reaction was modified on the electrode surface by chemical bonds via click chemistry. Then, N-acryloxysuccinimide (NAS)-labelled polymer chains were successfully formed on the electrode surface by ARGET-ATRP. Under the optimized conditions, a good linear relationship existed with the ECL signal and the logarithm of tRNA concentration in the range of 0.1 pM-10 nM, and the limit of detection was 2.61 fM. In addition, this strategy can identify mismatched bases and performs well in recovery assays in real samples. For its high sensitivity, selectivity, simplicity and economy, the ECL biosensor shows great potential for practical applications.


Subject(s)
Biosensing Techniques , Tobacco Mosaic Virus , Click Chemistry , Polymerization , RNA , Tobacco Mosaic Virus/genetics
20.
Anal Bioanal Chem ; 414(23): 6955-6964, 2022 Sep.
Article in English | MEDLINE | ID: mdl-35972525

ABSTRACT

Alkaline phosphatase (ALP), an important hydrolase involved in dephosphorylation, is a common clinical indicator of many diseases. In the present study, we constructed a novel electrochemical sensor using amifostine as the substrate of ALP and activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) as a signal amplification strategy for sensitive determination of ALP activity. In particular, in the presence of ALP, the phosphate group of amifostine was hydrolyzed to form a sulfhydryl group, which could attach to a gold electrode via a sulfur-gold bond. Then, the initiator α-bromophenylacetic acid (BPAA) was linked to the hydrolysis product of amifostine through an amide bond, resulting in the production of electroactive polymer chains on the gold electrode by the monomer ferrocenylmethyl methacrylate (FMMA) via ARGET ATRP. Under optimal parameters, the electrochemical sensor demonstrated a limit of detection (LOD) of 1.71 mU mL-1 with a linear range of 5-100 mU mL-1. In addition to satisfactory selectivity, the potential application of this approach for ALP activity detection in human serum samples was demonstrated. Due to its efficiency, simplicity of operation, and cost-effectiveness, the proposed electrochemical sensor has great promise as a universal method for ALP assays and inhibitor screening.


Subject(s)
Amifostine , Biosensing Techniques , Alkaline Phosphatase , Biosensing Techniques/methods , DNA/chemistry , Electrochemical Techniques/methods , Gold/chemistry , Humans , Limit of Detection
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