Detection Of Adenosine Triphosphate Research Articles

Non-enzymatic aptazyme-driven autonomous DNA circuits coupled with three-junction CHA for amplified live-cell ATP imaging

Live-cell imaging of adenosine triphosphate (ATP) is crucial for gaining insight into enzymatic activities and biological processes. However, it is still difficult to achieve sensitive and non-destructive ATP imaging due to the difficulty of amplifying molecules within cells and the complexity of intercellular environments. Herein, we present non-enzymatic ATP aptamer-based DNAzyme (Aptazyme)-driven autonomous DNA circuits for sensitive ATP imaging in living cells. The combination of dual amplifiers with aptazyme and catalysis hairpin assembly (CHA) circuits yielded a detection limit of 0.21 μM for ATP, which was 83.9 times more sensitive than the aptazyme-based probe without CHA circuits amplification. Moreover, the efficient carrier function and quenching effect of molybdenum disulfide (MoS2) made it possible to incorporate circuits components into cells without the requirement of transfection, thus allowing for efficient and non-destructive ATP detection within live cells. Therefore, the non-enzymatic aptazyme-driven autonomous DNA circuits present a promising novel approach for the sensitive detection of small molecules within live cells.

Magneto-fluorescent nanobiosensor with tetrahedral framework nucleic acid-confined aptamer for trace adenosine triphosphate detection

Accurate quantitation of trace-level ATP biomarkers often suffers from interference of sample matrix and remains a challenge. Herein, a novel magnetic biological nanosensor (MBNS) integrating tetrahedral framework nucleic acid (tFNA)-confined aptamers and ultrasensitive laser-induced fluorescence (LIF) was tailored and evaluated. The fluorescent aptamer nanoprobe hybridized with FAM-labelled cDNA was particularly incorporated into the inner cavity of tFNA, enabling a dual-mode mechanism that integrates molecular size selection and aptamer affinity recognition. By this strategy, the benefits of tFNA molecular sieve, the super sensitive properties of LIF, and the specific recognition of "aptamer gene codes" were integrated, as triggered the superior response of ATP. The structures, properties and reaction mechanism of tFNA, as well as tFNA modified with aptamer probe at different sites, were characterized in detail. A high level of anti-interference was achieved, shielding against non-specific adsorption even in matrices with high protein and analogue content, 103~105 times higher than ATP. The release of fluorescent beacon from aptamer probe was facilely driven by specific binding of ATP, and an ultra-sensitive detection of 75 pmol/L ATP was obtained without amplification. It was 2-3 orders of magnitude more sensitive than most fluorescent aptasensors. Applied to serum samples, the stability and reproducibility of the resultant aptasensor have been validated with the intra- and inter-assay RSD in 1.6~6.4% and -1.5~8.4%, respectively. Acceptable recoveries of 96.5~106.8% were achieved, consistent with the LC-MS method. It was expected to open a simple and efficient protocol for highly specific and ultrasensitive ATP detection in clinical diagnostics.

A Microfluidic Chip-Based Automated System for Whole-Course Monitoring the Drug Responses of Organoids.

Tumor patients-derived organoids, as a promising preclinical prediction model, have been utilized to evaluate ex vivo drug responses for formulating optimal therapeutic strategies. Detecting adenosine triphosphate (ATP) has been widely used in existing organoid-based drug response tests. However, all commercial ATP detection kits containing the cell lysis procedure can only be applied for single time point ATP detection, resulting in the neglect of dynamic ATP variations in living cells. Meanwhile, due to the limited number of viable organoids from a single patient, it is impractical to exhaustively test all potential time points in search of optimal ones. In this work, a multifunctional microfluidic chip was developed to perform all procedures of organoid-based drug response tests, including establishment, culturing, drug treatment, and ATP monitoring of organoids. An ATP sensor was developed to facilitate the first successful attempt on whole-course monitoring the growth status of fragile organoids. To realize a clinically applicable automatic system for the drug testing of lung cancer, a microfluidic chip based automated system was developed to perform entire organoid-based drug response test, bridging the gap between laboratorial manipulation and clinical practices, as it outperformed previous methods by improving data repeatability, eliminating human error/sample loss, and more importantly, providing a more accurate and comprehensive evaluation of drug effects.

Engineering smart thioflavin T binders with switchable DNA topologies for constructing multiple label-free biosensors

Thioflavin T (ThT) has been used as a sensitive fluorescence probe for lighting up nucleic acids. The binding of ThT is dependent on the structural topologies of DNA sequences. To date, most oligonucleotides interact with ThT through only a single topology, and this limits the applications of ThT as a sensing agent. Here, we modulated an HIV-1 integrase aptamer, 93del, to have switchable topologies by optimizing its 3′ flanking residues to enhance its ThT-binding capacity. Our data revealed that the sequences formed stem-loop hairpins and I-motif/G-quadruplex (G4) hybrids in Na+ buffer under alkaline and acidic conditions, respectively. Both topologies could bind ThT with high affinity (Kd ∼ 0.5 μM) and dramatically enhance the fluorescent signal of ThT. However, in the presence of K+, the sequences could be assembled into interlocked G4s, which had a low affinity for ThT but a high affinity for N-methyl mesoporphyrin IX (NMM). This allowed us to build highly sensitive fluorescent K+ sensors that were insusceptible to interference from Na+. Additionally, we found that the stacking of homologous base pairs in the hairpin loops was essential for ThT recognition and could be used as an indicator in the construction of label-free molecular beacons, allowing monitoring of microRNAs effectively in human serum spiked test. By employing external stimuli to induce the transformation of these topologies, we were able to sensitively detect adenosine triphosphate (ATP) molecules or Hg2+in vitro under specific acidic conditions. Our study provides a strategy for exploiting undiscovered functions of existing aptamers.

Anchoring Au nanoclusters into coordination polymers: A novel approach toward ATP detection and its application

Adenosine triphosphate (ATP) is the main source of energy required for all life activities and is used as a biomarker for diseases such as cancer. It is of great significance to design a novel fluorescent probe with favorable performance for monitoring the changes of ATP concentration. Herein, a fluorescence probe named ZnCPs@AuNCs for ATP sensing was designed and fabricated by integrating AuNCs into ZnCPs. The emission intensity of AuNCs was greatly enhanced upon the formation of the ZnCPs@AuNCs nanocomposites, which may be attributed to ZnCPs restricting the molecular motion of AuNCs. Upon the introduction of ATP, the fluorescence intensity at 564 nm of ZnCPs@AuNCs is quenched. According to this phenomenon, a sensitive and reliable ATP sensing platform was established. Moreover, ZnCPs@AuNCs were incorporated into a poly (vinyl alcohol) matrix for the fabrication of fluorescent film, which exhibited solid-state fluorescence. Inspired by the remarkable fluorescent properties of ZnCPs@AuNCs, the fluorescent hydrogel was prepared by mixing ZnCPs@AuNCs with κ-carrageenan, which demonstrated a response to ATP and favorable self-healing ability. This work presents a perspective of ZnCPs@AuNCs in multiple applications such as biosensing, fluorescent film, and hydrogel construction.

Nanopore single molecule ATP detection based on the dual effects of specific capturing and signal amplification

Adenosine triphosphate (ATP) plays a crucial role in energy metabolism. Some serious diseases such as Alzheimer’s disease are related to abnormal ATP concentrations. Fast, sensitive, and specific ATP detection is important and challenging in disease diagnosis and biomedical research. In present study, conical nanopore was designed and employed to detect ATP at single molecule level. For overcoming the inherent defects such as non-selectivity and low sensitivity of nanopore in sensing small target molecule, two collaborative improvements of specific identification and signal amplification were proposed. We first used split aptamer capture probes (H1 and S1) to anchor ATP molecule, and then used auxiliary probes (H2 and H3) to form a long-nicked duplex ATP-DNA concatemer by hybridization chain reaction (HCR). Referring ATP-DNA concatemer rather than native ATP to nanopore sensor, characteristic current signals with high signal-to-noise ratio were obtained. By such indirect way, specific and sensitive ATP detection was realized with a linear range from 20 pM to 5 nM and a detection limit of 3.9 pM (S/N = 3). Additionally, the sensor exhibited many merits such as simplify, anti-interference, and especially allowing ATP detection in real sample. So, we believe that the nanopore sensor has good application prospects in screening and management of ATP-related diseases.

Electrochemiluminescent quenching of luminol–doped Zr–MOFs for ATP detection through scavenging ROS using gallic acid–capped Au nanoparticles

In the present study, the highly efficient electrochemiluminescence (ECL) luminophores of Zr-based metal–organic frameworks (MOFs) coordinating with the amino group of luminol (Zr–Lu–MOFs) were developed for fabricating a sensing platform. The ECL emission of a Zr–Lu–MOF/H2O2 system with strong and stable ECL signals was triggered through cyclic voltammetry. Adenosine triphosphate (ATP) was firmly assembled on Zr–Lu–MOF because of the high affinity of Zr4+ toward phosphate groups. ATP was detected using gallic acid (GA)-capped Au (GA@Au) nanoparticles combined with an ATP aptamer. The GA@Au nanoparticles quenched the ECL behavior of the luminol/H2O2 system, and the quenching mechanism was ascribed to the GA scavenging superoxide radical (O2•−) that was generated via the electron redox reaction in H2O2. Under optimal experiments, an ECL biosensor with a linear range of 0.100–1000 nM and a detection limit of 0.0353 nM (S/N = 3) was designed for the selective and sensitive detection of ATP. Thus, the proposed ECL biosensor exhibited good performance with high stability and acceptable fabrication reproducibility, providing a valuable strategy for clinical diagnostics and therapeutics.

Realizing Ultrasensitive and Accurate Point-of-Care Profiling for ATP with a Triple-Mode Strategy Based on the ATP-Induced Reassembly of a Copper Coordination Polymer Nanoflower.

New strategies for accurate and reliable detection of adenosine triphosphate (ATP) with portable devices are significant for biochemical analysis, while most recently reported approaches cannot satisfy the detection accuracy and independent of large instruments simultaneously, which are unsuitable for fast, simple, and on-site ATP monitoring. Herein, a unique, convenient, and label-free point-of-care sensing strategy based on novel copper coordination polymer nanoflowers (CuCPNFs) was fabricated for multimode (UV-vis, photothermal, and RGB values) onsite ATP determination with high selectivity, sensitivity, and accuracy. The resulting CuCPNFs with a 3D hierarchical structure exhibit the ATP-triggered decomposition behavior because the competitive coordination between ATP and the copper ions of CuCPNFs can result in the formation of ATP-Cu, which reveals preeminent peroxidase mimics activity and can accelerate the oxidation of 3, 3', 5, 5'-tetramethylbenzidine (TMB) to form oxTMB. During this process, the detection system displayed not only color changes but also a strong NIR laser-driven photothermal effect. Thus, the photothermal and color signal variations are easily monitored by a portable thermometer and a smartphone. This multimode point-of-care platform can meet the requirements of onsite, without bulky equipment, accuracy, and reliability all at once, greatly enhancing its application in practice and paving a new way in ATP analysis.

Pyrene excimer-based fluorescent chemosensor for cascade detection of Zn(II) and phosphate ions and its applications

There is growing research on developing fluorescent chemosensors that can detect multiple analytes. Herein, a pyrene-naphthaldehyde based Schiff base PNY was synthesized, characterized, and employed for the cascade fluorescent ‘Off-On-Off’ detection of Zn2+ and phosphate ions. The interaction of PNY with Zn2+ led to a distinct fluorescence enhancement at 450 nm, and the fluorescent color of PNY was changed from blue to cyan-blue due to the complexation-induced formation of a pyrene excimer. PNY, as a fluorescent turn-on sensor can be employed to detect Zn2+ down to 2.20 × 10−7 M. Subsequently, the in-situ formed PNY-Zn2+ complex gets decomplex upon the addition of phosphate ion among other tested anions with a detection limit of 3.42 × 10−7 M. With the addition of phosphate ion, fluorescence intensity of PNY-Zn2+ at 450 nm was quenched with the concomitant formation of an emission peak at around 396 nm. The excimer-monomer mechanism was used to explain the sensing ability of PNY. Further, to complement the phosphate ion selectivity, the ability of PNY-Zn2+ to detect adenosine triphosphate (ATP) was explored. At last, the analytical novelty of PNY and PNY-Zn2+ was explored by quantifying Zn2+ and phosphate ions in real environmental and biological samples.

Development of a rhodamine-based fluorescent probe for ATP detection for potential applications in meat freshness assessment

Adenosine triphosphate (ATP) plays a vital role in physiological processes and is an essential indicator of microbial content in food. Herein, a new sensitive, rapid and water-soluble probe for ATP detection was developed. Rhodamine B and pentaethylenehexamine were employed to design and synthesise the probe rhodamine-pentaethylenehexamine (RP) for selective ATP detection. The synthesised probe RP was characterized using Fourier transform infrared, NMR and dynamic light scattering size distributions. Upon the addition of ATP, the probe exhibited a distinct change in fluorescence intensity, with fluorescence emission at 580 nm. A linear relationship was observed between fluorescence intensity and ATP concentrations at 0–50 μmol/L, with a limit of detection of 10.97 × 10−9 mol/L. The results of the zeta potential and molecular dynamics simulation demonstrated that the detection mechanism of the probe RP is associated with the electrostatic adsorption interaction between the multi-positively charged sites of RP and the negatively charged triphosphate structure of ATP. Our study provides new insights into improving charge site identification in small molecule detection. Furthermore, the successful detection of ATP on meat surfaces indicates that RP has the potential to assess meat freshness.

Organic photoelectrochemical transistor based on cascaded DNA network structure modulated ZnIn2S4/MXene Schottky junction for sensitive ATP detection

Organic photoelectrochemical transistor (OPECT) biosensor is now appearing in perspective of public, which characterized by amplified the grating electrode potential by ion transport. In this study, the DNA network formed by the hybridization chain reaction (HCR) detects the target adenosine triphosphate (ATP) by adjusting the surface potential of the new heterojunction of ZnIn2S4/MXene. The formation of DNA network amplifies the detection signal of ATP. Significantly, OPECT biosensor could further amplify the signal, which calculated the gain achieved 103, which is consistent with the gain signal of the previously reported OPECT biosensor. Furthermore, the OPECT biosensor achieved a highly sensitivity detection of the target ATP, which the linear detection range is 0.03 pM–30 nM, and the detection limit is 0.03 pM, and illustrated a high selectivity to ATP. The proposed OPECT biosensor achieved signal amplification by adjusting the surface potential of ZnIn2S4/MXene through cascade DNA network, which provides a new direction for the detection of biomolecules.

Modulation by Co (II) Ion of Optical Activities of L/D-glutathione (GSH)-modified Chiral Copper Nanoclusters for Sensitive Adenosine Triphosphate Detection.

Chiral nanoscale enantiomers exhibit different biological effects in living systems. However, their chirality effect on the detection sensitivity for chiral biological targets still needs to be explored. Here, we discovered that Co2+ can modulate the luminescence performance of L/D-glutathione (GSH)-modified copper nanoclusters (L/D-Cu NCs) and induce strong chiroptical activities as the asymmetric factor was enhanced 223-fold with their distribution regulating from the ultraviolet to visible region. One Co2+ coordinated with two GSH molecules that modified on the surface of Cu NCs in the way of CoN2O2. On this basis, dual-modal chiral and luminescent signals of Co2+ coordinated L/D-Cu NCs (L/D-Co-Cu NCs) were used to detect the chiral adenosine triphosphate (ATP) based on the competitive interaction between surficial GSH and ATP molecules with Co2+. The limits of detection of ATP obtained with fluorescence and circular dichroism intensity were 9.15 μM and 15.75 nM for L-Co-Cu NCs, and 5.35 μM and 4.69 nM for D-Co-Cu NCs. This demonstrated that selecting suitable chiral configurations of nanoprobes effectively enhances detection sensitivity. This study presents not only a novel method to modulate and enhance the chiroptical activity of nanomaterials but also a unique perspective of chirality effects on the detection performances for bio-targets.

Modulation by Co (II) Ion of Optical Activities of L/D‐glutathione (GSH)‐modified Chiral Copper Nanoclusters for Sensitive Adenosine Triphosphate Detection

AbstractChiral nanoscale enantiomers exhibit different biological effects in living systems. However, their chirality effect on the detection sensitivity for chiral biological targets still needs to be explored. Here, we discovered that Co2+ can modulate the luminescence performance of L/D‐glutathione (GSH)‐modified copper nanoclusters (L/D−Cu NCs) and induce strong chiroptical activities as the asymmetric factor was enhanced 223‐fold with their distribution regulating from the ultraviolet to visible region. One Co2+ coordinated with two GSH molecules that modified on the surface of Cu NCs in the way of CoN2O2. On this basis, dual‐modal chiral and luminescent signals of Co2+ coordinated L/D−Cu NCs (L/D−Co−Cu NCs) were used to detect the chiral adenosine triphosphate (ATP) based on the competitive interaction between surficial GSH and ATP molecules with Co2+. The limits of detection of ATP obtained with fluorescence and circular dichroism intensity were 9.15 μM and 15.75 nM for L−Co−Cu NCs, and 5.35 μM and 4.69 nM for D−Co−Cu NCs. This demonstrated that selecting suitable chiral configurations of nanoprobes effectively enhances detection sensitivity. This study presents not only a novel method to modulate and enhance the chiroptical activity of nanomaterials but also a unique perspective of chirality effects on the detection performances for bio‐targets.

Macrocyclic Conformational Switch Coupled with Pyridinium-Induced PET for Fluorescence Detection of Adenosine Triphosphate.

The binding of nucleotides is crucial for signal transduction as it induces conformational protein changes, leading to downstream cellular responses. Synthetic receptors that bind nucleotides and transduce the binding event into global conformational rearrangements are highly challenging to design, especially those that operate in an aqueous solution. Much work is focused on evaluating functionalized dyes to detect nucleotides, whereas coupling of a nucleotide-induced conformational switching to a sensing event has not been reported to date. We disclose synthetic receptors that undergo a global conformational rearrangement upon nucleotide binding. Integrating naphthalimide and the pyridinium ion into the structure enables stabilization of the folded conformation and efficient fluorescence quenching. The binding of a nucleotide rearranges the receptor conformation and alters the strong fluorescence enhancement. The methylpyridinium-containing receptor demonstrated high sensing selectivity for adenosine 5'-triphosphate (ATP) and a record 160-fold fluorescence enhancement. It can detect fluctuations of ATP in HeLa cells and possesses low cytotoxicity. The developed systems present an attractive approach for designing ATP-responsive artificial molecular switches that operate in water and integrate a strong fluorescence response.

Prognostic gene biomarkers for c-Src inhibitor Si162 sensitivity in melanoma cells.

Early detection and treatment are crucial in combating malignant melanoma. Src is an important therapeutic target in melanoma due to its association with cancer progression. However, developing effective Src-targeting drugs remains challenging and personalized medicine relies on biomarkers and targeted therapies for precise and effective treatment. This study focuses on Si162, a newly synthesized c-Src inhibitor, to identify reliable biomarkers for predicting Si162 sensitivity and explore associated biological characteristics and pathways in melanoma cells. Primary melanoma cells (M1, M21, M24, M84, M133, M307, and M2025) were obtained from patients diagnosed with melanoma. Si162 cytotoxicity tests were performed using luminescent adenosine triphosphate detection and the half-maximal inhibitory concentration (IC50) values were calculated. Gene expression profiles were analyzed using microarray-based gene expression data. Differentially expressed genes between the resistant and sensitive groups were identified using Pearson correlation analysis. Gene coexpression, interactions, and pathways were investigated through clustering, network, and pathway analyses. Biological functions were examined using the Database for Annotation, Visualization, and Integrated Discovery. Molecular pathways associated with different responses to Si162 were identified using gene set enrichment analysis. The gene expressions were validated using reverse transcription-quantitative polymerase chain reaction. The cells revealed significant differences in response to Si162 based on the IC50 values (p < 0.05). A total of 36 differentially expressed genes associated with Si162 susceptibility were identified. Distinct expression patterns between the sensitive and resistant groups were observed in 9 genes (LRBA, MGMT, CAND1, ADD1, SETD2, CNTN6, FGF18, C18orf25, and RPL13). Coexpression among the differentially expressed genes was highlighted, and 9 genes associated with molecular pathways, including EMT, transforming growth factor-beta (TGF-β) signaling, and ribosomal protein synthesis, between groups. Genes involved in dysregulated immune response were observed in the resistant group. The involvement of 5 genes (ADD1, CNTN6, FGF18, C18orf25, and RPL13) in Si162 resistance was confirmed through qRT-PCR validation. These findings contribute to our understanding of the underlying biological differences among melanoma cells and suggest potential biomarkers and pathways associated with Si162 response and resistance.

Biosensor for ATP detection via aptamer-modified PDA@POSS nanoparticles synthesized in a microfluidic reactor

This study introduces aptamer-functionalized polyhedral oligomeric silsesquioxane (POSS) nanoparticles for adenosine triphosphate (ATP) detection where the POSS nanoparticles were synthesized in a one-step, continuous flow microfluidic reactor utilizing thermal polymerization. A microemulsion containing POSS monomers was generated in the microfluidic reactor which was designed to prevent clogging by using a continuous oil flow around the emulsion during thermal polymerization. Surfaces of POSS nanoparticles were biomimetically modified by polydopamine. The aptamer sequence for ATP was successfully attached to POSS nanoparticles. The aptamer-modified POSS nanoparticles were tested for affinity-based biosensor applications using ATP as a model molecule. The nanoparticles were able to capture ATP molecules successfully with an affinity constant of 46.5 μ\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\upmu $$\\end{document}M. Based on this result, it was shown, for the first time, that microfluidic synthesis of POSS nanoparticles can be utilized in designing aptamer-functionalized nanosystems for biosensor applications. The integration of POSS in biosensing technologies not only exemplifies the versatility and efficacy of these nanoparticles but also marks a significant contribution to the field of biorecognition and sample preparation.

A Controlled Release Aptasensor Utilizing AIE-Active MOFs as High-Efficiency ECL Nanoprobe for the Sensitive Detection of Adenosine Triphosphate.

Improving the sensitivity in electrochemiluminescence (ECL) detection systems necessitates the integration of robust ECL luminophores and efficient signal transduction. In this study, we report a novel ECL nanoprobe (Zr-MOF) that exhibits strong and stable emission by incorporating aggregation-induced emission ligands into Zr-based metal-organic frameworks (MOFs). Meanwhile, we designed a high-performance signal modulator through the implementation of a well-designed controlled release system with a self-on/off function. ZnS quantum dots (QDs) encapsulated within the cavities of aminated mesoporous silica nanoparticles (NH2-SiO2) serve as the ECL quenchers, while adenosine triphosphate (ATP) aptamers adsorbed on the surface of NH2-SiO2 through electrostatic interaction act as "gatekeepers." Based on the target-triggered ECL resonance energy transfer between Zr-MOF and ZnS QDs, we establish a coreactant-free ECL aptasensor for the sensitive detection of ATP, achieving an impressive low detection limit of 0.033 nM. This study not only demonstrates the successful combination of ECL with controlled release strategies but also opens new avenues for developing highly efficient MOFs-based ECL systems.

Portable and Recyclable Luminescent Lanthanide Coordination Polymer Film Sensors for Adenosine Triphosphate in Urine.

Adenosine triphosphate (ATP) is a small molecule that is released to the urine from bladder urothelial cells and the bladder mucosal band of the human body. In certain cases, ATP can serve as a biomarker in bladder disease. For the practical applicability of luminescent sensors for ATP in urine, it is significant to find a new strategy for making the detection progress simple and available for in-field urine analysis. Here, a novel luminescent lanthanide coordination polymer (Tb-BPA) was designed and synthesized for quick and sensitive detection of ATP through luminescence quenching with a quenching constant of 4.90 × 103 M-1 and a detection limit of 0.55 × 10-6 M. Besides, Tb-BPA has excellent anti-interference ability and can detect ATP in simulated urine with a small relative standard deviation (<4%). Moreover, the luminescent polyacrylonitrile nanofiber films modified by Tb-BPA were prepared by electrospinning and were used for ATP visual detection. Notably, this film is easy to recover and reuse, and maintains good detection performance after at least 7 cycles.

Dual-signal viscosity flow paper sensor for ATP detection based on bio-recognition and nanozyme activity regulation of ZIF-90

Adenosine triphosphate (ATP) is an important biological biomarker, but its rapid extracellular decomposition poses challenges for on-site detection. We report a paper-based dual-readout viscosity flow sensor enabling rapid, equipment-free quantification of ATP. The assay integrates molybdenum disulfide nanozymes for signal amplification with zeolitic imidazolate framework-90 (ZIF-90) for selective ATP recognition and nanozyme regulation. In the presence of ATP, the intact chitosanase-encapsulated ZIF-90 structure was disintegrated to release chitosanase and thus hydrolyzed chitosan hydrogel, decreasing the viscosity of chitosan solution. Moreover, elimination of ZIF-90 inhibitory effect reactivated the catalytic activity of MoS2 nanozyme, resulting in the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to produce obvious color change. The analytical signals, combining the distance- and colorimetric- readout, were directly displayed on filter paper without further processed. The proposed dual-readout viscosity flow sensor exhibited remarkable sensitivity for ATP detection with detection limits down to 0.038 nM with the distance-readout and 0.057 nM for the colorimetric-readout. Furthermore, we developed an integrated paper kit with a printed scale and color card for standardized on-site analysis without instrumentation. Successful application for ATP quantification in bacterial biofilms and blood samples demonstrates the sensor’s strong performance and potential for decentralized testing.

Evaluation of ATP bioluminescence for rapid determination of cleanliness of livestock trailers after a commercial wash.

Pathogens such as porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome (PRRSV), and E. coli are known to spread by contaminated vehicles and equipment. Pork producers have adopted trailer wash policies where each trailer is washed, disinfected, and dried before it can return to a farm. Cleanliness of livestock trailers after washing is determined by visual inspection rather than any objective method. Adenosine triphosphate (ATP) bioluminescence is used in many industries to provide real-time feedback on surface cleanliness through the detection of ATP from organic sources. That same technology may provide trailer wash facilities a way of objectively characterizing a livestock trailer's suitability to return to a farm after washing. Two ATP luminometers (3M Clean-Trace and Neogen AccuPoint) were used to estimate the correlation between ATP bioluminescence readings and aerobic bacterial plate counts (APCs) from sampled surfaces and to determine locations within a livestock trailer that can accurately estimate surface cleanliness. Five locations in livestock trailers were evaluated. Those locations included the nose access door (NAD), back door flush gate, rear side access door (RSAD), belly flush gate (BFG), and belly side access door (BSAD). There was a positive log-log association between the two luminometers (r = 0.59, P < 0.01). Every log unit increase in one unit, resulted in a 0.42 log increase (P < 0.01) in the other unit. ATP can come from bacteria, yeasts, molds, and manure. There was a poor association (r ≥ 0.10, P ≥ 0.02) between APCs and the ATP luminometers. Still, an increase in relative light units (RLUs) resulted in a corresponding increase in colony-forming units. The greatest area of surface contamination measured by APC was the NAD. RLUs were also greater in the NAD compared to the RSAD, the BFG, and the BSAD (P ≤ 0.01). Because APCs and luminometer RLUs provided similar outcomes, statistical process control charts were developed to determine control limits for RLUs. This provides real-time feedback to trailer wash workers in determining cleanliness outcomes for livestock trailers. These data suggest that ATP bioluminescence can be a reliable method to monitor cleaning effectiveness in livestock trailers. Bioluminescence is a monitoring tool that should be used in conjunction with microbial methods to monitor procedures for cleaning and disinfection.