3D Film-Like Nanozyme with a Synergistic Amplification Effect for the Ultrasensitive Immunochromatographic Detection of Respiratory Viruses.

Greatly improving the sensitivity and detection range of lateral flow immunoassays (LFAs) by at least 100 times without using additional instruments remains challenging. Herein, we develop a three-dimensional (3D) film-like nanozyme (GO-Pt30-AuPt5) by ordered assembly of one layer of 30 nm Pt nanoparticles (NPs) and one layer of small Au@Pt satellites (5 nm) onto a two-dimensional (2D) graphene oxide (GO) nanofilm, in which GO greatly increased the interface area and stability of the nanozyme whereas Pt and Au@Pt NPs synergistically enhanced colorimetric/catalytic activities. The grafting of outer Au@Pt satellites converted the 2D nanofilm into a 3D flexible nanozyme with numerous catalytic sites for enzymatic deposition signal amplification and binding sites for target capture. The introduction of GO-Pt30-AuPt5 into multiplex LFA achieved the ultrasensitive and simultaneous detection of two important respiratory viruses with sensitivity of 1 pg/mL level, which was about 100 times higher than that without signal enrichment and at least 20 and 1900 times higher than those of traditional enzyme-linked immunosorbent assay and AuNP-based LFA, respectively. The clinical utility of the proposed assay was validated through the diagnosis of 49 real clinical respiratory tract specimens. Our proposed LFA shows great potential for the ultrasensitive screening of pathogens in the field.

Engineering the bacteriophage 80 alpha endolysin as a fast and ultrasensitive detection toolbox against Staphylococcus aureus.

The isolation and identification of pathogenic bacteria from a variety of samples are critical for controlling bacterial infection-related health problems. The conventional methods, such as plate counting and polymerase chain reaction-based approaches, tend to be time-consuming and reliant on specific instruments, severely limiting the effective identification of these pathogens. In this study, we employed the specificity of the cell wall-binding (CBD) domain of the Staphylococcus aureus bacteriophage 80 alpha (80α) endolysin towards the host bacteria for isolation. Amidase 3-CBD conjugated magnetic beads successfully isolated as few as 1 × 102 CFU/mL of S. aureus cells from milk, blood, and saliva. The cell wall hydrolyzing activity of 80α endolysin promoted the genomic DNA extraction efficiency by 12.7 folds on average, compared to the commercial bacterial genomic DNA extraction kit. Then, recombinase polymerase amplification (RPA) was exploited to amplify the nuc gene of S. aureus from the extracted DNA at 37 °C for 30 min. The RPA product activated Cas12a endonuclease activity to cleave fluorescently labeled ssDNA probes. We then converted the generated signal into a fluorescent readout, detectable by either the naked eye or a portable, self-assembled instrument with ultrasensitivity. The entire procedure, from isolation to identification, can be completed within 2 h. The simplicity and sensitivity of the method developed in this study make it of great application value in S. aureus detection, especially in areas with limited resource supply.

Enzyme- and label-free cascade isothermal amplification aptasensor for the ultrasensitive detection of ochratoxin A.

BACKGROUND: Ultrasensitive detection is crucial for the early warning and intervention of risk factors, ultimately benefiting the environment and human health. Low levels of ochratoxin A (OTA) present a hidden yet significant threat, and rapid detection via high-performing biosensors is therefore essential. RESULTS: A cascade isothermal amplification aptasensor (CIA-aptasensor) was designed for OTA detection. On the surface of a magnetic bead probe, the OTA level was converted into positively correlated trigger cDNA through its competitive binding with OTA-Apt. The released trigger cDNA activated catalytic hairpin assembly followed by coupling with a hybridization chain reaction to achieve CIA. After adding graphene oxide and SYBR Green I, the background interference was eliminated to specifically obtain OTA-related fluorescence. The ultrasensitive limit of detection was 0.22 pg mL-1, an improvement of 1368-fold over conventional enzyme-linked aptamer sorbent assay by the same OTA-Apt, demonstrating satisfactory reliability and practicability. Thus, the CIA-aptasensor provides an enzyme- and label-free simplified homogeneous system with minimal background interference using isothermal conditions. SIGNIFICANCE: This study provides a polymerase chain reaction-like approach for enhancing the sensitivity and performance of a biosensor, which could be extended for the application of CIA and label-free signaling strategy to other risk factors.

An ultrasensitive dual-mode stagey for 17ß-estradiol assay: Photoelectrochemical and colorimetric biosensor based on a WSe2/TiO2-modified electrode coupled with nucleic acid amplification.

BACKGROUND: The abuse of 17ß-estradiol(E2) has aroused wide concern in environmental and biomedical fields, which severely affects the endocrine function of human and animals. Therefore, an ultrasensitive and accurate assay of E2 is critically important. Traditional chromatography or immunoassay techniques exhibited good sensitivity and selectivity, but expensive instruments and antibodies may pose cost and stability issues, as well as difficulties in meeting on-site detection requirements. Ultrasensitive, reliable, and on-site detection of E2 at trace level remains a challenge. Hence, developing a simple, ultrasensitive assay to simultaneously achieve accurate detection and rapid visual analysis of E2 is extremely crucial. RESULTS: We developed a versatile dual-mode photoelectrochemical (PEC) and colorimetric biosensor based on isothermal nucleic acid amplification strategy for the ultrasensitive and accurate detection of E2. The method modified titanium dioxide (TiO2) with tungsten selenide (WSe2) nanoflowers to synthesize WSe2/TiO2 heterostructures as a substrate for signal amplification and nanoprobe modification. Isothermal nucleic acid amplification strategy has been proven to be a powerful tool for strong signal amplification. The presence of a target triggered the nucleic acid amplification reaction, and produced a large amount of tDNA that competed with G-quadruplex immobilized on the electrode surface. The remaining G-quadruplex/hemin catalyzed the 4-chloro-1-naphthol (4-CN) to form biocatalytic precipitation (BCP) and ABTS-H2O2 chromogenic reaction, thus, the dual-mode platform was capable of achieving PEC-colorimetric ultrasensitive detection based on the catalytic activity of G-quadruplex/hemin DNAzyme. Within optimal conditions, the dual-mode biosensor exhibited a remarkable detection limit as low as 0.026 pM. SIGNIFICANCE: Benefiting from the superior performance of WSe2/TiO2 and the power signal amplification of isothermal nucleic acid amplification strategy, this aptasensor achieved the ultrasensitive detection of E2. The independent transmission paths of photoelectrochemical and colorimetric provide mutual support and flexible switching, significantly enhancing the overall sensitivity and accuracy of the detection strategy, which can meet the needs for E2 precise quantification and rapid on-site detection.

Employing conductive porous hydrogen-bonded organic framework for ultrasensitive detection of peanut allergen Ara h1.

Peanut allergy has garnered worldwide attention due to its high incidence rate and severe symptoms, stimulating the demand for the ultrasensitive detection method of peanut allergen. Herein, we successfully developed a novel electrochemical aptasensor for ultrasensitive detection Ara h1, a major allergenic protein present in peanuts. A conductive nickel atoms Anchored Hydrogen-Bonded Organic Frameworks (PFC-73-Ni) were utilized as excellent electrocatalysts toward hydroquinone (HQ) oxidation to generate a readable current signal. The developed electrochemical aptasensor offers wide linear range (1-120 nM) and low detection limit (0.26 nM) for Ara h1. This method demonstrated a recovery rate ranging from 95.00% to 107.42% in standard addition detection of non-peanut food samples. Additionally, the developed electrochemical method was validated with actual samples and demonstrated good consistency with the results obtained from a commercial ELISA kit. This indicates that the established Ara h1 detection method is a promising tool for peanut allergy prevention.

A Novel Terahertz Metamaterial Microfluidic Sensing Chip for Ultra-Sensitive Detection.

A terahertz metamaterial microfluidic sensing chip for ultrasensitive detection is proposed to investigate the response of substances to terahertz radiation in liquid environments and enhance the molecular fingerprinting of trace substances. The structure consists of a cover layer, a metal microstructure, a microfluidic channel, a metal reflective layer, and a buffer layer from top to bottom, respectively. The simulation results show that there are three obvious resonance absorption peaks in the range of 1.5-3.0 THz and the absorption intensities are all above 90%. Among them, the absorption intensity at M1 = 1.971 THz is 99.99%, which is close to the perfect absorption, and its refractive index sensitivity and Q-factor are 859 GHz/RIU and 23, respectively, showing excellent sensing characteristics. In addition, impedance matching and equivalent circuit theory are introduced in this paper to further analyze the physical mechanism of the sensor. Finally, we perform numerical simulations using refractive index data of normal and cancer cells, and the results show that the sensor can distinguish different types of cells well. The chip can reduce the sample pretreatment time as well as enhance the interaction between terahertz waves and matter, which can be used for early disease screening and food quality and safety detection in the future.

Ultra-Sensitive Detection of the SARS-CoV-2 Nucleocapsid Protein via a Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a-Mediated Immunoassay.

Tracking trace protein analytes in precision diagnostics is an ongoing challenge. Here, we developed an ultrasensitive detection method for the detection of SARS-CoV-2 nucleocapsid (N) protein by combining enzyme-linked immunosorbent assay (ELISA) with the clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) system. First, the SARS-CoV-2 N protein bound by the capture antibody adsorbed on the well plate was sequentially coupled with the primary antibody, biotinylated secondary antibody, and streptavidin (SA), followed by biotin primer binding to SA. Subsequently, rolling circle amplification was initiated to generate ssDNA strands, which were targeted by CRISPR/Cas12a to cleave the FAM-ssDNA-BHQ1 probe in trans to generate fluorescence signals. We observed a linear relationship between fluorescence intensity and the logarithm of N protein concentration ranging from 3 fg/mL to 3 × 107 fg/mL. The limit of detection (LOD) was 1 fg/mL, with approximately nine molecules in 1 µL of the sample. This detection sensitivity was 4 orders magnitude higher than that of commercially available ELISA kits (LOD: 5.7 × 104 fg/mL). This method was highly specific and sensitive and could accurately detect SARS-CoV-2 pseudovirus and clinical samples, providing a new approach for ultrasensitive immunoassay of protein biomarkers.

Construction of AlGaN/GaN high-electron-mobility transistor-based biosensor for ultrasensitive detection of SARS-CoV-2 spike proteins and virions.

The COVID-19 pandemic has highlighted the need for rapid and sensitive detection of SARS-CoV-2. Here, we report an ultrasensitive SARS-CoV-2 immunosensor by integration of an AlGaN/GaN high-electron-mobility transistor (HEMT) and anti-SARS-CoV-2 spike protein antibody. The AlGaN/GaN HEMT immunosensor has demonstrated the capability to detect SARS-CoV-2 spike proteins at an impressively low concentration of 10-22 M. The sensor was also applied to pseudoviruses and SARS-CoV-2 ΔN virions that display the Spike proteins with a single virion particle sensitivity. These features validate the potential of AlGaN/GaN HEMT biosensors for point of care tests targeting SARS-CoV-2. This research not only provides the first HEMT biosensing platform for ultrasensitive and label-free detection of SARS-CoV-2.

Development of surface-enhanced Raman scattering-sensing Method by combining novel Ag@Au core/shell nanoparticle-based SERS probe with hybridization chain reaction for high-sensitive detection of hepatitis C virus nucleic acid.

The ultrasensitive detection of hepatitis C virus (HCV) nucleic acid is crucial for the early diagnosis of hepatitis C. In this study, by combining Ag@Au core/shell nanoparticle (Ag@AuNP)-based surface-enhanced Raman scattering (SERS) tag with hybridization chain reaction (HCR), a novel SERS-sensing method was developed for the ultrasensitive detection of HCV nucleic acid. This SERS-sensing system comprised two different SERS tags, which were constructed by modifying Ag@AuNP with a Raman reporter molecule of 4-ethynylbezaldehyde, two different hairpin-structured HCR sequences (H1 or H2), and a detection plate prepared by immobilizing a capture DNA sequence onto the Ag@AuNP layer surface of the detection wells. When the target nucleic acid was present, the two SERS tags were captured on the surface of the Ag@AuNP-coated detection well to generate many "hot spots" through HCR, forming a strong SERS signal and realizing the ultrasensitive detection of the target HCV nucleic acid. The limit of detection of the SERS-sensing method for HCV nucleic acid was 0.47 fM, and the linear range was from 1 to 105 fM.

Ultrasensitive Flexible Temperature Sensors Based on Thermal-Mediated Ions Migration Dynamics in Asymmetrical Polymer Bilayers.

Accurately acquiring crucial data on the ambient surroundings and physiological processes delivered via subtle temperature fluctuation is vital for advancing artificial intelligence and personal healthcare techniques but is still challenging. Here, we introduce an electrically induced cation injection mechanism based on thermal-mediated ion migration dynamics in an asymmetrical polymer bilayer (APB) composed of nonionic polymer and polyelectrolyte layers, enabling the development of ultrasensitive flexible temperature sensors. The resulting optimized sensor achieves ultrahigh sensitivity, with a thermal index surpassing 10,000 K-1, which allows identifying temperature differences as small as 10 mK with a sensitivity that exceeds 1.5 mK. The mechanism also enables APB sensors to possess good insensitivity to various mechanical deformations─features essential for practical applications. As a proof of concept, we demonstrate the potential impact of APB sensors in various conceptual applications, such as mental tension evaluation, biomimetic thermal tactile, and thermal radiation detection.

Aggregation-induced signal amplification strategy based on peptide self-assembly for ultrasensitive electrochemical detection of melanoma biomarker.

The detection of melanoma circulating biomarker in liquid biopsies is current under evaluation for being potentially utilized for earlier cancer diagnosis and its metastasis. Herein, we developed a non-invasive electrochemical approach for ultrasensitive detection of the S100B, serving as a potential promising blood circulating biomarker of melanoma, based on an aggregation-induced signal amplification (AISA) strategy via in-situ peptide self-assembly. The fundamental principle of this assay is that the designed amphiphilic peptides (C16-Pep-Fc), fulfilling multiple functions, feature both a recognition region for specific binding to S100B and an aggregation (self-assembly) region for the formation of peptide nanomicelles under mild conditions. The C16 tails were encapsulated within the hydrophobic core of the aggregates, while the relatively hydrophilic recognition fragment Pep and Fc tag were exposed on the outer surface for subsequent recognition of S100B and signal output. AISA provided remarkable accumulation of electroactive Fc moieties that enabled ultrasensitive S100B detection of as low as 0.02 nM, which was 10-fold lower than un-amplified approach and better than previously reported assays. As a proof-of-concept study, further experiments also highlighted the good reproducibility and stability of AISA and demonstrated its usability when applied to simulated serum samples. Hence, this work not only presented a valuable assay tool for ultrasensitive detecting protein biomarker, but also advocated for the utilization of aggregation-induced signal amplification in electrochemical biosensing system, given its considerable potential for future practical applications.

A Near-Infrared Fluorogenic Probe for Rapid, Specific, and Ultrasensitive Detection of Sphingosine in Living Cells and In Vivo.

Sphingosine (Sph) plays important roles in various complex biological processes. Abnormalities in Sph metabolism can result in various diseases, including neurodegenerative disorders. However, due to the lack of rapid and accurate detection methods, understanding sph metabolic in related diseases is limited. Herein, a series of near-infrared fluorogenic probes DMS-X (X = 2F, F, Cl, Br, and I) are designed and synthesized. The fast oxazolidinone ring formation enables the DMS-2F to detect Sph selectively and ultrasensitively, and the detection limit reaches 9.33 ± 0.41 nm. Moreover, it is demonstrated that DMS-2F exhibited a dose- and time-dependent response to Sph and can detect sph in living cells. Importantly, for the first time, the changes in Sph levels induced by Aß42 oligomers and H2 O2 are assessed through a fluorescent imaging approach, and further validated the physiological processes by which Aß42 oligomers and reactive oxygen species (ROS)-induce changes in intracellular Sph levels. Additionally, the distribution of Sph in living zebrafish is successfully mapped by in vivo imaging of a zebrafish model. This work provides a simple and efficient method for probing Sph in living cells and in vivo, which will facilitate investigation into the metabolic process of Sph and the connection between Sph and disease pathologies.

Highly selective and sensitive histamine and tryptamine analysis using SiO2@AuNPs@PDA molecularly imprinted polymer coupled with SALDI-TOF MS.

A combination of SiO2@AuNPs@PDA molecularly imprinted and surface-assisted laser desorption/ionization-time-of-flight mass spectrometry (SALDI-TOF MS) was devised as a method for highly specific and ultrasensitive detection of two biogenic amines-histamine (HIS) and tryptamine (TRP)-in real samples. In this strategy, AuNPs modified amino-abundant silica nanospheres (SiO2@AuNPs). The prepared SiO2@AuNPs were used as a substrate to synthesize a molecularly imprinted polymer (MIP) through in situ dopamine self-polymerization with HIS and TRP as the template molecules (SiO2@AuNP@PDA-MIP). The as-prepared MIP structure, properties, and target-analyte identification conditions were characterized and optimized and it was used as the matrix for MS. Compared to the case of nonimprinted materials, the imprinting function endowed the matrix with a higher selectivity for capturing the target molecules. The enriched analytes were directly and rapidly identified using SALDI-TOF MS without elution. Meanwhile, the proposed method has low background interference, good reproducibility and stability, high salt tolerance, and satisfactory linearity (R2 > 0.99), and it enables ultrasensitive detection of HIS and TRP (limits of detection for HIS and TRP were 0.2 and 0.1 ng mL-1, respectively). Moreover, the proposed method was applied to analyze samples of real beer, sausage, and chicken, and the results agreed with those obtained via liquid chromatography-MS, suggesting that the method has excellent practical applications in the field of food safety.

Ultrasensitive fluorescence detection of gonyautoxins in seawater using a novel molecularly imprinted nanoprobe.

Gonyautoxins (GTXs), a group of potent neurotoxins belonging to paralytic shellfish toxins (PSTs), are often associated with harmful algal blooms of toxic dinoflagellates in the sea and represent serious health and ecological concerns worldwide. In the study, a highly selective and sensitive fluorescence nanoprobe was constructed based on photoinduced electron transfer recognition mechanism to rapidly detect GTXs in seawater, using specific entrapment of molecularly imprinted polymers (MIPs) combined with fluorescence analyses. The green emissive fluorescein isothiocyanate was grafted in a silicate matrix as a signal transducer and fluorescence intensity of the nanoprobe with a core-shell structure exhibited a strong enhancement due to efficient analyte blockage in a short response time. Under optimal conditions, the developed MIPs nanoprobe presented an excellent analytical performance for spiked seawater samples including a recovery from 94.44 % to 98.23 %, a linear range between 0.018 nmol L-1 and 0.36 nmol L-1, as well as good accuracy. Furthermore, the method had extremely high sensitivity, with limit of detection obtained as 0.005 nmol L-1 for GTXs and GTX2/3. Finally, the nanoprobe was applied for the determination of GTXs in seven natural seawater samples with GTXs mixture (0.035-0.058 nmol L-1) or single GTX2/3 (0.033-0.050 nmol L-1), and the results agreed well with those of a UPLC-MS/MS method. The findings of our study suggest that the constructed MIPs-based fluorescence enhancement nanoprobe was suitable for rapid, selective and ultrasensitive detection of GTXs, particular GTX2/3, in natural seawater samples.

Self-Assembled Three-Dimensional Polyamide/Silver Nanoparticle Pore Array as a Highly Sensitive and Reproducible SERS Substrate for Pesticide Detection in Water.

In this study, silver nanoparticles (AgNPs) are self-assembled onto the polyamide (PA) pore array through hydrogen bonding, resulting in and optimizing the PA/Ag 3D pore array substrates. The best surface-enhanced Raman scattering (SERS) substrate is obtained with a pore depth of 500 nm in the PA array, 30 nm AgNPs, at a pH of 5.0, and a 24 h assembly time. The SERS performance of the substrates is assessed using rhodamine 6G (R6G) as a probe molecule. The detection limit of the R6G molecule reaches 10-13 M, and the relative standard deviation is under 20%, indicating good enhancement ability and reproducibility. Furthermore, label-free detection of pesticide contaminant diquat with a detection limit of 2.69 × 10-9 M is achieved using the optimized 3D substrate, which meets environmental monitoring requirements for drinking water. The findings demonstrate that this 3D SERS substrate has promising potential for use and development in the fields of contaminant detection and chemical sensing.

Development of antibody-aptamer sandwich-like immunosensor based on RCA and Nicked-PAM CRISPR/Cas12a system for the ultra-sensitive detection of a biomarker.

Biomarkers are the most sensitive reactants and early indicators of many kinds of diseases. The development of highly sensitive and simple techniques to quantify them is challenging. In this study, based on rolling cycle amplification (RCA) and the Nicked PAM/CRISPR-Cas12a system (RNPC) as a signal reporter, a sandwich-type method was developed using antibody@magnetic beads and aptamer for the high-sensitive detection of the C-reactive protein (CRP). The antibody-antigen (target)-aptamer sandwich-like reaction was coupled to RCA, which can produce hundreds of similar binding sites and are discriminated by CRISPR/Cas12a for signal amplification. The ultrasensitivity is achieved based on the dual-signal enhancing strategy, which involves the special recognition of aptamers, RCA, and trans-cleavage of CRISPR/Cas12a. By incorporating the CRISPR/Cas12a system with cleaved PAM, the nonspecific amplification of the RCA reaction alone was greatly reduced, and the dual signal output of RCA and Cas12a improved the detection sensitivity. Our assay can be performed only in two steps. The first step takes only 20 min of target capture, followed by a one-pot reaction, where the target concentration can be obtained by fluorescence values as long as there are 37 °C reaction conditions. Under optimal conditions, this system detected CRP with high sensitivity. The fabricated biosensor showed detection limits of 0.40 pg/mL in phosphate-buffered saline and 0.73 pg/mL in diluted human serum and a broad linear dynamic range of 1.28 pg/mL to 100 ng/mL within a total readout time of 90 min. The method could be used to perform multi-step signal amplification, which can help in the ultrasensitive detection of other proteins. Overall, the proposed biosensor might be used as an immunosensor biosensor platform.

CRISPR/Cas-Powered Amplification-Free Detection of Nucleic Acids: Current State of the Art, Challenges, and Futuristic Perspectives.

CRISPR/Cas system is becoming an increasingly influential technology that has been repositioned in nucleic acid detection. A preamplification step is usually required to improve the sensitivity of CRISPR/Cas-based detection. The striking biological features of CRISPR/Cas, including programmability, high sensitivity and sequence specificity, and single-base resolution. More strikingly, the target-activated trans-cleavage could act as a biocatalytic signal transductor and amplifier, thereby empowering it to potentially perform nucleic acid detection without a preamplification step. The reports of such work are on the rise, which is not only scientifically significant but also promising for futuristic end-user applications. This review started with the introduction of the detection methods of nucleic acids and the CRISPR/Cas-based diagnostics (CRISPR-Dx). Next, we objectively discussed the pros and cons of preamplification steps for CRISPR-Dx. We then illustrated and highlighted the recently developed strategies for CRISPR/Cas-powered amplification-free detection that can be realized through the uses of ultralocalized reactors, cascade reactions, ultrasensitive detection systems, or others. Lastly, the challenges and futuristic perspectives were proposed. It can be expected that this work not only makes the researchers better understand the current strategies for this emerging field, but also provides insight for designing novel CRISPR-Dx without a preamplification step to win practicable use in the near future.

Development of a novel fluorescent aptasensor based on the interaction between hexagonal ß-Co(OH)2 nanoplates and nitrogen-doped carbon dots for ultrasensitive detection of patulin.

There is an urgent need to develop an economical and convenient method for the ultrasensitive detection of patulin (PAT), a mycotoxin that can potentially harm human health when it is found in fruits and their derivatives. In this study, we have developed a novel fluorescent aptasensor that utilizes nitrogen-doped carbon dots (N-CDs) as the fluorescent donor and hexagonal ß-Co(OH)2 nanoplates as the fluorescent acceptor. N-CDs were synthesized through the hydrothermal method, resulting in spherical particles with a diameter of 7.6 nm. These nanoparticles exhibited excellent water solubility and displayed a vibrant blue emission at 448 nm when excited at 360 nm. Cobalt hydroxide nanoplates with a beta crystal structure [ß-Co(OH)2] were synthesized using a simple co-precipitation method, exhibiting hexagonal plate-like shapes with uniform lateral sizes of 4-5 µm. The fluorescence of N-CDs can be efficiently quenched by hexagonal ß-Co(OH)2 nanoplates through Förster resonance energy transfer mechanism. The maximum quenching-recovery capability can be achieved when the concentrations of N-CDs-Apt and ß-Co(OH)2 nanoplates are 150 nmol/L and 100 µg/mL, respectively. The pH of the TE buffer should be 8.0, and the incubation time should be 10 min at 25 °C. The developed fluorescent aptasensor displayed an excellent selectivity for PAT determination with a detection limit of 0.57 pg/mL in the linear range of 1.25 pg/mL-100 ng/mL. The rapid PAT determination in fruit juice samples was realized with good recoveries (96.9-105.8%). The developed fluorescent aptasensor based on the interaction between N-CDs and hexagonal ß-Co(OH)2 nanoplates can be a promising method for the rapid and ultrasensitive detection of PAT in agricultural products.

Triple strategy-enhanced immunochromatographic assay based on APCB and AIEFM for the ultrasensitive detection of AFM1.

Aflatoxin M1 (AFM1) is highly toxic, widely distributed, and difficult to monitor, posing a serious threat to human health. Therefore, a highly sensitive, rapid, convenient, and low-cost detection method must be urgently established. In this study, a triple strategy-enhanced immunochromatographic assay (ICA) was developed to satisfy these detection requirements. First, a turn-on signal output mode of the fluorescence quenching ICA substituted the turn-off mode of the traditional ICA for sensitive response to trace AFM1, with the limit of detection (LOD) reduced by approximately 4.9-fold. Then, a novel Au and polydopamine (PDA) cogrowth chrysanthemum-like blackbody was prepared as the quenching probe to reduce the background signal. This probe combined the excellent properties of Au nanoparticles with PDA. Thus, its fluorescence quenching constant was higher than that of single Au and PDA nanoparticles by 25.8- and 4.9-fold, respectively. Furthermore, an aggregation-induced emission fluorescence microsphere with a 5.7-fold higher relative quantum yield than a commercial fluorescence microsphere was selected as the signal output carrier to improve the signal-to-noise ratio. The integration of the above triple strategies established a 53.4-fold sensitivity-enhanced fluorescence quenching ICA (LOD = 0.9 pg/mL) for detecting AFM1 in milk, providing a strong technical guarantee for the safety monitoring of milk products.

Colorimetric immunosensing using liposome encapsulated MnO2 nanozymes for SARS-CoV-2 antigen detection.

Development of specific signal reporters with signal amplification effect are highly needed for sensitive and accurate detection of pathogen. Herein, we design a colorimetric immunosensing nanosystem based on liposome encapsulated quantum dots-sized MnO2 nanozyme (MnO2QDs@Lip) as a signal reporter for ultrasensitive and fast detection of SARS-CoV-2 antigen. The pathogenic antigens captured and separated by antibody-conjugated magnetic beads (MBs) are further connected with antibody-modified MnO2QDs@Lip to form a sandwich-like immunocomplex structure. After triggered release, MnO2 QDs efficiently catalyze colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue oxidized TMB, which can be qualitatively observed by naked eyes and quantitatively analyzed by UV-Vis spectra or smartphone platforms. By taking advantages of immuno-magnetic separation, excellent peroxidase-like catalytic activity of MnO2 QDs, and high encapsulation efficiency of MnO2QDs@Lip, ultrasensitive detection of SARS-CoV-2 antigen ranging from 0.1 pg/mL to 100 ng/mL is achieved within 20 min. The limit of detection (LOD) is calculated to be 65 fg/mL in PBS buffer. Furthermore, real clinical samples of SARS-CoV-2 antigens can be effectively identified by this immunosensing nanosystem with excellent accuracy. This proposed detection nanosystem provides a strategy for simple, rapid and ultrasensitive detection of pathogens and may shed light on the development of new POCT detection platforms for early diagnosis of pathogens and surveillance in public health.