Continuous in vivo monitoring of biomarkers remains challenging due to limited sensitivity, integration, and biocompatibility. Here, we report an integrated microneedle-based electrochemiluminescence device (MN-ECLD) for real-time detection of protein biomarkers in interstitial fluid. Leveraging hydrogen-bonded organic frameworks with ultrabright, biocompatible electrochemiluminescence, the emitters were incorporated into porous gold-coated microneedle arrays and regulated via interface-specific Y-shaped probes, enabling efficient coreactant-free signal generation. The device achieved ultrasensitive protein detection in vitro with a linear range of 100 fg/mL to 10 ng/mL, a detection limit of 21.3 fg/mL, and stability over 12 days, delivering an 87-fold sensitivity enhancement over conventional emitters. In vivo, MN-ECLD enabled real-time monitoring of cardiac biomarkers, achieving early warning of acute myocardial infarction in rats and pigs, with biomarker trends consistent with serum ELISA. This work establishes a versatile platform for continuous in vivo diagnostics of acute cardiovascular and metabolic disorders.

Personalized health management aims to promote, maintain, and restore the health of individuals. Despite the ever-lasting research efforts involved in personalized healthcare bioelectronics, current healthcare platforms still face barriers such as costly facilities, specialized operations, and resource-limited applications. Therefore, personalized and user-friendly healthcare bioelectronics are urgently needed. Among emerging solutions, the integration of artificial intelligence (AI) and advanced bioelectronics is a pivotal approach that merges intelligent algorithms with multi-functional healthcare design. This review summarizes the latest advances in AI-assisted bioelectronics, aiming to provide a possible strategy for personalized healthcare applications. Initially, a brief survey is provided to discuss the material design, device fabrication, AI-hardware integration, and performance assessment of AI-assisted bioelectronics. The subsequent contents focus on the implementation of AI-assisted healthcare bioelectronics across health monitoring, early diagnosis, therapeutic treatment, and rehabilitation. Finally, we discuss the current challenges and prospective future developments in closed-loop healthcare bioelectronics, ultimately empowering individuals with control over their own health.

Nucleic acid amplification tests are widely used for molecular diagnostics. However, primer-dimer formation and nonspecific amplification may compromise analytical accuracy and increase the risk of false-positive results. Here, temperature-gated PNA@AuNRs (peptide nucleic acid-modified gold nanorods) were introduced to improve amplification specificity. A microwave-assisted modification method efficiently immobilized PNA onto AuNRs, enabling directional primer capture at low temperature and controlled release near polymerase-optimal conditions. PNA@AuNRs-LAMP achieved a limit of quantification (LOQ) of 102 copies/μL with strong linearity (R2 = 0.991), while PNA@AuNRs-qPCR reached 10 copies/μL with excellent linearity (R2 = 0.998). Mechanistic analysis demonstrated strong PNA-primer affinity and a distinct thermal-start effect at 60 °C, absent in traditional LAMP. In spiked equine serum and nasal samples, PNA@AuNRs effectively eliminated nonspecific amplification and improved detection accuracy. These results highlight PNA@AuNRs-gated amplification as a promising strategy for high-specificity molecular diagnostics.

Advancing clinical diagnostics requires platforms that combine catalytic efficiency, biocompatibility, and real-time, in vivo accessibility. Herein, this study reports a structurally integrated FePc-ZIF-8-MX nanozyme that combines the redox activity of FePc, the porous confinement of ZIF-8, and the electrical conductivity of MX. Synthesized via a low-energy, ambient-condition process, this hybrid enables efficient electron transfer, enhanced analyte enrichment, and sustained catalytic activity in physiological environments. To translate this functionality into a wearable diagnostic format, the hybrid is seamlessly incorporated into a microneedle array, offering minimally invasive access to interstitial fluid for continuous L-cysteine (L-Cys) monitoring. The resulting platform exhibits high selectivity and sensitivity across complex biological matrices, including serum, urine, cultured cells, and a murine model of myocardial infarction. This study presents a multifunctional electrochemical platform that enables on-body metabolite monitoring through a microneedle-integrated nanozyme interface. To the best of our knowledge, it constitutes the first realization of real-time, in vivo L-Cys sensing in this format, setting a new benchmark for precision biosensing in translational healthcare.

Wearable theranostics hold great promise in precision medicine and real-time monitoring of diseases, which are capable of performing both predictive analysis and therapeutics concurrently. This review, for the first time, provides a detailed description of wearable device applications in theranostic studies, including smart contact lenses, smart bandages, smart dressing, and wearable theranostic dental patch, as well as the practical use of wearable therapeutic devices in clinical settings. In addition, wearable technologies enhanced by machine learning techniques, which are capable of enhancing the precision of theranostics through autonomous learning in early stage diagnosis and treatment, image analysis, and predictive analytics, are analyzed. Third, the situation of commercialization of these bioelectronics is summarized. Finally, the existing challenges and future directions for translation and commercialization of the wearable theranostics are discussed in detail. AI-assisted wearable theranostic systems are transitioning from laboratory innovations to clinical applications, enabling intelligent and convenient clinical translation and deployment models, as a paradigm shift in modern personalized medicine.

Loop-mediated isothermal amplification (LAMP) holds great promise for rapid nucleic acid detection. However, its application is hindered by the notable occurrence of false positives. In this work, gold nanorod (AuNR)-mediated hot start effect, which adsorbs primers and controls the release of single-stranded DNA (ssDNA) primers at about 50 °C, was demonstrated. The AuNR-mediated LAMP reaction for detecting African horse sickness virus (AHSV) showed high selectivity against other DNA fragments, even at high magnesium concentrations. Furthermore, the assay achieves excellent sensitivity with a detection limit of 100 copies/μL. Additionally, the method displays good repeatability from 50 to 65 °C (optimal), and the coefficient of variation of Tt at the same concentration of nucleic acids is less than 5%. Our study optimizes the LAMP reaction using AuNRs, thereby reducing false positives and offering great potential for clinical diagnosis.

Continuous measurement of macromolecular biomarkers in vivo could enable diverse implantable applications in personalized medicine. However, technical obstacles remain: current technologies are limited to only one-way tracking of increases or decreases in macromolecule levels and lack real-time feedback on disease progression. Here, we propose an integrated diagnosis-therapy sensing system for dynamic tracking of cell-free DNA and drug delivery, based on a semi-implantable indwelling needle modified with clustered regularly interspaced short palindromic repeats (CRISPR)-dCas9. The specific binding-dissociation sensing mechanism of CRISPR-dCas9 with target DNA on the surface under fluctuating blood flow is discussed in detail, with various reaction equilibrium constants. Owing to the matched mechanical properties and geometrical structure of the semi-implantable device, it shows the ability to withstand interference of 60 % fetal bovine serum, sensitivity of 300 fM, 3-day stability, and real-time feedback on target DNA level in animal models. For sepsis patients bearing Staphylococcus aureus, the biosensor exhibited clinical sensitivity and specificity of 92.3 % and 100 % respectively. It could enable dynamic monitoring of cell-free DNA in vivo and timely intervention for patients with acute syndromes such as sepsis in the intensive care unit (ICU).

Real-time and continuous monitoring of nucleic acid biomarkers with wearable devices holds potential for personal health management, especially in the context of pandemic surveillance or intensive care unit disease. However, achieving high sensitivity and long-term stability remains challenging. Here, we report a tetrahedral nanostructure-based Natronobacterium gregoryi Argonaute (NgAgo) for long-term stable monitoring of ultratrace unamplified nucleic acids (cell-free DNAs and RNAs) in vivo for sepsis on wearable device. This integrated wireless wearable consists of a flexible circuit board, a microneedle biosensor, and a stretchable epidermis patch with enrichment capability. We comprehensively investigate the recognition mechanism of nucleic acids by NgAgo/guide DNA and signal transformation within the Debye distance. In vivo experiments demonstrate the suitability for real-time monitoring of cell-free DNA and RNA with a sensitivity of 0.3 fM up to 14 days. These results provide a strategy for highly sensitive molecular recognition in vivo and for on-body detection of nucleic acid.

In this experiment, we present a microfluidic-based molecular fluorescence spectroscopy method for analyzing nucleic acids to identify transgenic soybeans. This method is integrated into a General Chemistry Experiment course tailored for freshmen. The primary goals of this course are to deepen student's understanding of some important knowledge points of general chemistry, introduce students to experimental techniques at the micro- and nanoscale, help students understand the principles of molecular fluorescence spectroscopy and enzyme reaction kinetics, elucidate the relationship between chemistry and its practical applications, stimulate their interest in chemistry, and provide multidisciplinary perspectives and thinking. Over the period from 2021 to 2023, more than 180 freshmen enrolled in this course, and over 30 universities in China have initiated the advancement of this course. Most students successfully completed the experiment, achieving high completion rate and promising results. Participating students improved their practical skills and the ability to work collaboratively in a laboratory setting, which led to numerous innovative ideas and insights in related areas. Positive feedback from the students confirmed that the predetermined learning objectives were successfully achieved.

On-site diagnostic tests that accurately identify disease biomarkers lay the foundation for self-healthcare applications. However, these tests routinely rely on single-mode signals and suffer from insufficient accuracy, especially for multiplexed point-of-care tests (POCTs) within a few minutes. Here, this work develops a dual-mode multiclassification diagnostic platform that integrates an electrochemiluminescence sensor and a field-effect transistor sensor in a microfluidic chip. The microfluidic channel guides the testing samples to flow across electro-optical sensor units, which produce dual-mode readouts by detecting infectious biomarkers of tuberculosis (TB), human rhinovirus (HRV), and group B streptococcus (GBS). Then, machine-learning classifiers generate three-dimensional (3D) hyperplanes to diagnose different diseases. Dual-mode readouts derived from distinct mechanisms enhance the anti-interference ability physically, and machine-learning-aided diagnosis in high-dimensional space reduces the occasional inaccuracy mathematically. Clinical validation studies with 501 unprocessed samples indicate that the platform has an accuracy approaching 99%, higher than the 77%-93% accuracy of rapid point-of-care testing technologies at 100% statistical power (>150 clinical tests). Moreover, the diagnosis time is 5 min without a trade-off of accuracy. This work solves the occasional inaccuracy issue of rapid on-site diagnosis, endowing POCT systems with the same accuracy as laboratory tests and holding unique prospects for complicated scenes of personalized healthcare.

Prokaryotic Argonautes (pAgos) have been recently used in many nucleic acid biosensing applications but have rarely been used for regulating the isothermal amplification system. Herein, we reported Thermus thermophilus Argonaute (TtAgo)-mediated background-suppressed exponential isothermal amplification (EXPAR) as the first example to explore the binding activity of pAgos toward regulation of the amplification template. It was demonstrated that thermophilic pAgos efficiently eliminated nonspecific hybridization between templates by their binding affinity with the template, resulting in greatly enhancing the specificity of EXPAR. TtAgo-mediated, background-suppressed EXPAR was employed to detect miRNA with a detection limit of 10-15 M, which was 1000 times and 100 times more sensitive than that of traditional RT-PCR and EXPAR, respectively. This method further showed good performance in discriminating cancer patients from healthy individuals, indicating its potential for practical clinical applications.

In this paper,we designed a protein immunoassay using microfluidic technology to quantitatively detect procalcitonin(PCT).The experiment involved various steps such as solution preparation,microfluidic chip assembly,standard curve drawing,testing of unknown liquid samples,and other experimental processes.By conducting these experiments,students can gain a deeper understanding of the emerging scientific research field of microfluidics.Furthermore,this training helps to foster their scientific thinking,ability to innovate,and integrate knowledge.

IntroductionDifferent pathogens causing mixed infection are now threatening the pig industry in the context of the African Swine Fever (ASF) circulating especially in China, and it is crucial to achieving the early diagnosis of these pathogens for disease control and prevention.MethodsHere we report the development of a rapid, portable, sensitive, high-throughput, and accurate microfluidic-LAMP chip detection system for simultaneous detection and differentiation of gene-deleted type and wild-type African swine fever virus (ASFV), pseudorabie virus (PRV), porcine parvovirus (PPV), porcine circovirus type 2 (PCV2), and porcine reproductive and respiratory syndrome (PRRSV).Results and discussionThe newly developed system was shown to be sensitive with detection limits of 101 copies/μl for ASFV-MGF505-2R/P72, PPV, and PCV2, 102 copies/μl for ASFV-CD2v, PRV, and PRRSV. The system was highly specific (100%) and stable (C.V.s < 5%) in its ability to detect different pathogens. A total 213 clinical samples and 15 ASFV nucleic acid samples were collected to assess the performance of the detection system, showing highly effective diagnosis. Altogether, the developed microfluidic-LAMP chip system provides a rapid, sensitive, high-throughput and portable diagnostic tool for the accurate detection of multiple swine pathogens.

A cytokine storm may be the last attack of various diseases, such as sepsis, cancer, and coronavirus disease 2019, that can be life threatening. Real‐time monitoring of cytokines in vivo is helpful for assessing the immune status of patients and providing an early warning of a cytokine storm. In this study, a functional carbon nanotube biointerface‐based wearable microneedle patches for real‐time monitoring of a cytokine storm in vivo via electrochemical analysis are reported. This wearable system has sensitivity with a detection limit of 0.54 pg mL −1 , high specificity, and 5 days of stability with a coefficient of variation of 4.0%. The system also has a quick response of several hours (1–4 h) to increasing cytokines. This wearable microneedle patch may offer a promising route for real‐time biomolecule wearables construction. The patch is also the first reported integrated capture and monitoring system that is capable of real‐time measurement of protein markers in interstitial fluid.

The rapid and accurate detection of peanuts and soybeans allergen is important to the food safety.In this study,Cu-TCPP nanosheet,a kind of ultra-thin metal-organic framework(MOF)was synthesized and applied in loop-mediated isothermal amplification(named Cu-TCPP@LAMP),which can inhibit the non-specific amplification by absorbing and precise temperature releasing of single primer.As thus,Cu-TCPP@LAMP can achieve high sensitivity and specific amplification of the target gene.As a result,peanut and soybean allergens genes contained in food were successfully detected with a favorable detection sensitivity(5 ng/μL for peanuts and 10 ng/μL for soybeans)and reliable repeatability(The coefficient of variation was 3.38％for peanuts and 3.33％for soybeans).Moreover,the established method was utilized for detection of several commercial products,and had a high consistency with the standard method.Apart from food allergens,this novel assay can be widely used in other areas,such as pathogen detection,tumor nucleic acid detection and so on.

Sensitive, simple and visual detection methods have become the focus of research because they can realize realtime detection without rely on the various detection instruments, so it is of great significance to apply them to the detection of African swine fever virus (ASFV). In this work, functionalized bismuth quantum dots (Bi QDs) were prepared by hydrothermal method and used in loop-mediated isothermal amplification (LAMP) reaction to replace the fluorescent dyes. Due to the overlap of the spectra of Bi QDs and titanium carbide (Ti3C2) nanosheet, the fluorescence resonance energy transfer (FRET) phenomenon could occurred between the Bi QDs and Ti3C2 nanosheet, and Ti3C2 nanosheets were used as energy acceptor and Bi QDs were used as fluorescence donors, which resulted in the quench of Bi QDs fluorescence signal. When the target ASFV was present, the LAMP reaction was activated due to the specific binding between the primer and the target result the separation of the Bi QDs from the surface of the Ti3C2 nanosheets, which leading to fluorescence recovery. Thus a target-driven fluorescence recovery sensing system for ASFV detection was developed. The detection results of the sensing system showed that the visual blue fluorescence recovered gradually in the range of 5.0 x 10-12-1.0 x 10-6 g/L with the detection limit of 1.5 x 10-12 g/L (2.4 copies/mu L) and satisfactory detection results were obtained in the infected ASFV samples. The proposed fluorescent visual sensing detection strategy opens up a new way for sensitive, simple and visual nucleic acid detection.

Abstract Early diagnosis of acute diseases is restricted by the sensitivity and complex process of sample treatment. Here, an ultrasensitive, rapid, and portable electrochemiluminescence‐microfluidic (ECL‐M) system is described via sandwich‐type immunoassay and surface plasmonic resonance (SPR) assay. Using a sandwich immunoreaction approach, the ECL‐M system employs cardiac troponin‐I antigen (cTnI) as a detection model with a Ru@SiO2 NPs labeled antibody as the signal probe. For miR‐499‐5p detection, gold nanoparticles generate SPR effects to enhance Ru(bpy)32+ ECL signals. The system based on alternating current (AC) electroosmotic flow achieves an LOD of 2 fg mL−1 for cTnI in 5 min and 10 aM for miRNAs in 10 min at room temperature. The point‐of‐care testing (POCT) device demonstrated 100% sensitivity and 98% specificity for cTnI detection in 123 clinical serum samples. For miR‐499‐5p, it exhibited 100% sensitivity and 97% specificity in 55 clinical serum samples. Continuous monitoring of these biomarkers in rats' saliva, urine, and interstitial fluid samples for 48 hours revealed observations rarely documented in biotic fluids. The ECL‐M POCT device stands as a top‐performing system for ECL analysis, offering immense potential for ultrasensitive, rapid, highly accurate, and facile detection and monitoring of acute diseases in POC settings.

In this study, we developed an innovative highly specific nucleic acid isothermal detection assay based on prokaryotic DNA polymerase I with exquisitely designed fluorescent probes, achieving high sensitivity and 100% specificity within 30 min. The fluorescent nucleic acid probe was designed and constructed based on the specific flap cleavage endonuclease activity of prokaryotic DNA polymerase I (including the Bst, Bsu, Bsm, and Klenow DNA polymerases). The flap endonuclease activity depends on the length of the flap DNA and polymerization activity, which greatly reduces the false-positive rate caused by primer dimerization. This robust assay was also validated by the detection of rotavirus with great specificity and sensitivity. It could be a great alternative to qPCR in the field of point-of-care detection of pathogens.

In this study, we developed a fluorescent nucleic acid probe-based assay for the rapid and specific detection of nucleic acids, achieving high sensitivity and 100% specificity within 30 minutes. The nucleic acid probe was designed and constructed based on the specific flap cleavage endonuclease activity of prokaryotic DNA polymerase I (including the Bst, Bsu, Bsm, and Klenow DNA polymerases). The flap endonuclease activity depends on the length of the flap DNA and upon polymerization activity. This fluorescent probe could be valuable for the detection of nucleic acids, such as for accurate genetic analysis, or the rapid and point-of-care detection of pathogens.