Nucleic acid and nanomaterial-assisted signal-amplified strategies in fluorescent analysis of circulating tumor cells and small extracellular vesicles.

As two main types of liquid biopsy markers, both circulating tumor cells (CTCs) and small extracellular vesicles (sEVs) play important roles in the diagnosis and prognosis of cancers. CTCs are malignant cells that detach from the original tumor tissue and enter the circulation of body fluids. sEVs are nanoscale vesicles secreted by normal cells or pathological cells. However, CTCs and sEVs in body fluids are scarce, leading to great difficulties in the accurate analysis of related diseases. For the sensitive detection of CTCs and sEVs in body fluids, various types of nucleic acid and nanomaterial-assisted signal amplification strategies have been developed. In this review, we summarize the recent advances in fluorescent detection of CTCs and sEVs in liquid biopsy based on nucleic acid and nanomaterial-assisted signal amplification strategies. We also discuss their advantages, challenges, and future prospects.

Recent Advances in Biological Applications of Aptamer-Based Fluorescent Biosensors.

Aptamers have been spotlighted as promising bio-recognition elements because they can be tailored to specific target molecules, bind to targets with a high affinity and specificity, and are easy to chemically synthesize and introduce functional groups to. In particular, fluorescent aptasensors are widely used in biological applications to diagnose diseases as well as prevent diseases by detecting cancer cells, viruses, and various biomarkers including nucleic acids and proteins as well as biotoxins and bacteria from food because they have the advantages of a high sensitivity, selectivity, rapidity, a simple detection process, and a low price. We introduce screening methods for isolating aptamers with q high specificity and summarize the sequences and affinities of the aptamers in a table. This review focuses on aptamer-based fluorescence detection sensors for biological applications, from fluorescent probes to mechanisms of action and signal amplification strategies.

Nanomaterial-Mediated Immunosensors and Their Performance in Detecting Tumor Markers.

The detection of tumor markers is crucial for assessing the progression of specific cancers. Numerous research studies have shown that immunosensors can convert immune-specific response biosignals into visual signals, enabling the highly sensitive tracking and detection of tumor markers. This offers a promising solution for early cancer diagnosis. However, most tumor markers are inert molecules that are challenging to detect at low concentrations in the early stages of cancer. Therefore, there is a need to develop immunosensor analysis platforms with a higher sensitivity. Nanomaterials, with their advantages of high stability, low cost, and versatility in design, have emerged as ideal candidates for enhancing the performance of immunosensor analysis. In this paper, we review the design ideas of nanomaterials in antibody-based electrochemical, electrochemiluminescent, and photoelectrochemical immunosensors, including electrode interface modification, signaling probes for stimulating sensing signals, and design strategies of modified materials in signaling mechanisms. In addition, we have thoroughly analyzed the performance, advantages and disadvantages of different immunosensors. Therefore, the aim of this paper is to review the recent advances in advanced nanomaterial strategies for different immunosensors and their biomedical applications, and to point out the challenges and prospects of immunosensors in future clinical applications.

Signal amplification strategies in biosensing of extracellular vesicles (EVs).

Extracellular vesicles (EVs) are membrane-enclosed vesicles secreted from mammalian cells. EVs act as multicomponent delivery vehicles to carry a wide variety of biological molecular information and participate in intercellular communications. Since elevated levels of EVs are associated with some pathological states such as inflammatory diseases and cancers, probing circulating EVs holds a great potential for early diagnostics. To this end, several detection methods have been developed in which biosensors have attracted great attentions in identification of EVs due to their simple instrumentation, versatile design and portability for point-of-care applications. The concentrations of EVs in bodily fluids are extremely low (i.e. 1-100 per µl) at early stages of a disease, which necessitates the use of signal amplification strategies for EVs detection. In this way, this review presents and discusses various amplification strategies for EVs biosensors based on detection modalities including surface plasmon resonance (SPR), calorimetry, fluorescence, electrochemical and electrochemiluminescence (ECL). In addition, microfluidic systems employed for signal amplification are reviewed and discussed in terms of their design and integration with the detection methods.

RNA-Cleaving DNAzyme-Based Amplification Strategies for Biosensing and Therapy.

Since their first discovery in 1994, DNAzymes have been extensively applied in biosensing and therapy that act as recognition elements and signal generators with the outstanding properties of good stability, simple synthesis, and high sensitivity. One subset, RNA-cleaving DNAzymes, is widely employed for diverse applications, including as reporters capable of transmitting detectable signals. In this review, the recent advances of RNA-cleaving DNAzyme-based amplification strategies in scaled-up biosensing are focused, the application in diagnosis and disease treatment are also discussed. Two major types of RNA-cleaving DNAzyme-based amplification strategies are highlighted, namely direct response amplification strategies and combinational response amplification strategies. The direct response amplification strategies refer to those based on novel designed single-stranded DNAzyme, and the combinational response amplification strategies mainly include two-part assembled DNAzyme, cascade reactions, CHA/HCR/RCA, DNA walker, CRISPR-Cas12a and aptamer. Finally, the current status of DNAzymes, the challenges, and the prospects of DNAzyme-based biosensors are presented.

Aptamer-based signal amplification strategies coupled with microchips for high-sensitivity bioanalytical applications: A review.

With their advantages in specificity, high stability and easy screening, aptamers are becoming increasingly popular recognition elements for biosensor platforms. At the same time, microchips as the new analytical detection platforms have achieved significant growth in the past decades. At present, with the intersection of aptamer and microfluidic technology, aptamer-based high-sensitivity bioanalysis on microchips exhibits a great application potential in biomedical science and environmental fields. In this review, we highlight the recent progress in high-sensitivity bioanalytical applications based on aptamer signal amplification strategies on microchips. Furthermore, the main challenges in the practical application are discussed, and the development in the future is prospected.

Recent Progresses in Electrochemical DNA Biosensors for MicroRNA Detection.

MicroRNAs (miRNAs), as the small, non-coding, evolutionary conserved, and post-transcriptional gene regulators of the genome, have been highly associated with various diseases such as cancers, viral infections, and cardiovascular diseases. Several techniques have been established to detect miRNAs, including northern blotting, real-time polymerase chain reaction (RT-PCR), and fluorescent microarray platform. However, it remains a significant challenge to develop sensitive, accurate, rapid, and cost-effective methods to detect miRNAs due to their short size, high similarity, and low abundance. The electrochemical biosensors exhibit tremendous potential in miRNA detection because they satisfy feature integration, portability, mass production, short response time, and minimal sample consumption. This article reviewed the working principles and signal amplification strategies of electrochemical DNA biosensors summarized the recent improvements. With the development of DNA nanotechnology, nanomaterials and biotechnology, electrochemical DNA biosensors of high sensitivity and specificity for microRNA detection will shortly be commercially accessible.

Electrochemical Signal Amplification Strategies and Their Use in Olfactory and Taste Evaluation.

Biosensors are powerful analytical tools used to identify and detect target molecules. Electrochemical biosensors, which combine biosensing with electrochemical analysis techniques, are efficient analytical instruments that translate concentration signals into electrical signals, enabling the quantitative and qualitative analysis of target molecules. Electrochemical biosensors have been widely used in various fields of detection and analysis due to their high sensitivity, superior selectivity, quick reaction time, and inexpensive cost. However, the signal changes caused by interactions between a biological probe and a target molecule are very weak and difficult to capture directly by using detection instruments. Therefore, various signal amplification strategies have been proposed and developed to increase the accuracy and sensitivity of detection systems. This review serves as a reference for biosensor and detector research, as it introduces the research progress of electrochemical signal amplification strategies in olfactory and taste evaluation. It also discusses the latest signal amplification strategies currently being employed in electrochemical biosensors for nanomaterial development, enzyme labeling, and nucleic acid amplification techniques, and highlights the most recent work in using cell tissues as biosensitive elements.

Recent advances in catalytic hairpin assembly signal amplification-based sensing strategies for microRNA detection.

Accumulative evidences have indicated that abnormal expression of microRNAs (miRNAs) is closely associated with many health disorders, making them be regarded as potentialbiomarkers for early clinical diagnosis. Therefore, it is extremely necessary to develop a highly sensitive, specific and reliable approach for miRNA analysis. Catalytic hairpin assembly (CHA) signal amplification is an enzyme-free toehold-mediated strand displacement method, exhibiting significant potential in improving the sensitivity of miRNA detection strategies. In this review, we first describe the potential of miRNAs as disease biomarkers and therapeutics, and summarize the latest advances in CHA signal amplification-based sensing strategies for miRNA monitoring. We describe the characteristics and mechanism of CHA signal amplification and classify the CHA-based miRNA sensing strategies into several categories based on the "signal conversion substance", including fluorophores, enzymes, nanomaterials, and nucleotide sequences. Sensing performance, limit of detection, merits and disadvantages of these miRNA sensing strategies are discussed. Moreover, the current challenges and prospects are also presented.

Recent advances in DNA walker machines and their applications coupled with signal amplification strategies: A critical review.

DNA walkers, a type of dynamic nanomachines, have become the subject of burgeoning research in the field of biology. These walkers are powered by driving forces based on strand displacement reactions, protein enzyme/DNAzyme reactions and conformational transitions. With the unique properties of high directionality, flexibility and efficiency, DNA walkers move progressively and autonomously along multiple dimensional tracks, offering abundant and promising applications in biosensing, material assembly and synthesis, and early cancer diagnosis. Notably, DNA walkers identified as signal amplifiers can be combined with various amplification approaches to enhance signal transduction and amplify biosensor sensing signals. Herein, we systematically and comprehensively review the walking principles of various DNA walkers and the recent progress on multiple dimensional tracks by presenting representative examples and an insightful discussion. We also summarized and categorized the diverse signal amplification strategies with which DNA walkers have coupled. Finally, we outline the challenges and future trends of DNA walker machines in emerging analytical fields.

A dual-model "on-super off" photoelectrochemical/ratiometric electrochemical biosensor for ultrasensitive and accurate detection of microRNA-224.

A dual-model "on-super off" photoelectrochemical (PEC)/ratiometric electrochemical (EC) biosensor based on signal enhancing and quenching combining three-dimensional (3D) DNA walker strategy was designed for the ultrasensitive and accurate detection of microRNA-224 (miRNA-224). The "signal on" PEC state was achieved by methylene blue labeled hairpin DNA (MB-DNA) for sensitizing CdS QDs. Then numerous transformational ferrocene labeled DNAs (Fc-DNAs) converted by target-induced 3D DNA walker amplification with the help of Ag nanocubes (NCs) label DNA (Ag-DNA) were introduced to open hairpin MB-DNA. Such configuration change would relocate the sensitizer MB and the quencher Fc, whereas energy transfer placed between Ag NCs and CdS QDs, thereby significantly quenching the PEC signal to obtain "super off" state. Meanwhile, these changes resulted in a decreased oxidation peak current of MB (IMB) and an increased that of Fc (IFc). MiRNA-224 was also detected on basis of the dual-signaling EC ratiometric method for complementary PEC detection. Benefiting from different mechanisms and relatively independent signal transduction, this approach not only avoided interference from difficult assembly but also outstandingly increased sensitivity by distance-controllable signal enhancing and quenching strategies. As a result, the detection ranges of 0.1-1000 fM with a low detection limit of 0.019 fM for PEC, and 0.52 to 500 fM with a low detection limit of 0.061 fM for EC, were obtained for miRNA-224, which opens a new avenue for designing numerous elegant biosensors with potential utility in bioanalysis and early disease diagnosis.

Advanced sensitivity amplification strategies for voltammetric immunosensors of tumor marker: State of the art.

Immunosensors are molecular recognition-based solid-state biosensing devices, in which the immunochemical reactions are coupled with transducers. As biologic or biochemical substances produced by tumor cells, tumor marker plays an important role in clinical diagnosis and treatment of cancer because its concentration is related to tumor size, clinical stage, and predicting prognosis. Voltammetric immunosensors based on the electrochemical analysis technique provide a sensitive electroanalytical approach for quantitatively detecting tumor markers by measuring the current as a function of the potential. To satisfy the need for accurate monitoring of tumor markers in low-concentration and their slight changes in concentration, the primary aim of developing a novel voltammetric immunosensor is to improve its sensitivity and limit of detection. Compared with traditional immunoassay, the advanced sensitivity-amplified immunosensors have applied appropriate amplification strategies to convert the bio-signal of antigen-antibody recognition events to the high electrochemical signal of redox species. Building on the significant concepts, sensitivity and limit of detection, we describe how the performance of voltammetric immunosensors can be improved by various sensitivity amplification mechanisms: (1) construction of labels with a high loading of signal species; (2) introduction of interfacial reaction initiated by functionalized nanomaterials; (3) building a synergistic connection between labels and substrate. The review ends with a summary of the shortage of current sensitivity amplified immunosensors and the perspective of enhancement strategies for more simple, efficient, and reliable voltammetric immunosensors.

Recent advances in duplex-specific nuclease-based signal amplification strategies for microRNA detection.

In recent years, miRNAs had emerged as promising biomarkers for diagnosis and prognosis of the health-threatening diseases (e.g., cancers), which associated with a broad range of pathological and biological processes, including drug resistance, apoptosis, metastasis, and proliferation. Therefore, accurate detection of the levels of miRNA shown excellent prospects for early diagnosis of the health-threatening diseases. Considering that only trace miRNA existed in biological fluids, many newly developed biosensors for miRNA detection mainly focused on introducing various of signal amplification strategies for improving the detection sensitivity. Duplex-specific nuclease (DSN), a nuclease purified from hepatopancreas of Kamchatka crab, was capable of specifically cleaving double-stranded DNA or DNA in DNA-RNA heteroduplexes and was inactive toward single-stranded oligonucleotides or double-stranded RNA, endowing a possibility for construction of newly miRNA biosensors. Recently, many newly developed DSN-based biosensors architectures for miRNA analysis were reported. In this review, we explained the great potential of miRNAs as promising biomarkers by overviewing DSN-based signal amplification strategies for miRNA detection in the last decades.

A sandwich-type electrochemical aptasensor for Mycobacterium tuberculosis MPT64 antigen detection using C60NPs decorated N-CNTs/GO nanocomposite coupled with conductive PEI-functionalized metal-organic framework.

The present work described a novel sandwich-type electrochemical aptasensor for rapid and sensitive determination of Mycobacterium tuberculosis MPT64 antigen. Herein, a novel carbon nanocomposite composed of fullerene nanoparticles, nitrogen-doped carbon nanotubes and graphene oxide (C60NPs-N-CNTs/GO) was facilely synthesized for the first time, which not only possessed a large specific surface area and excellent conductivity, but also exhibited outstanding inherent electroactive property, and therefore served as nanocarrier and redox nanoprobe simultaneously. Gold nanoparticles (AuNPs) was then uniformly anchored onto the surface of such nanocomposite via Au-N bonds to bind with MPT64 antigen aptamer Ⅱ (MAA Ⅱ), forming the tracer label to realize generation and amplification of electrochemical signal. Additionally, conductive polyethyleneimine (PEI)-functionalized Fe-based metal-organic framework (P-MOF) was used as a sensing platform to absorb bimetallic core-shell Au-Pt nanoparticles (Au@Pt), which could accelerate electron transfer and increase the immobilization of MPT64 antigen aptamer Ⅰ (MAA Ⅰ). After the typical sandwich-type protein-aptamer recognition, the inherent electroactivity of the tracer label was provoked by tetraoctylammonium bromide (TOAB), leading to a well-defined current response. Under the optimum condition, the proposed aptasensor showed a wide linear range for MPT64 detection from 1 fg/mL to 1 ng/mL with a limit of detection (LOD) as low as 0.33 fg/mL. More importantly, it was successfully used for MPT64 antigen detection in human serum, exhibiting a promising prospect for TB diagnosis in clinical practice.

Research advances in the detection of miRNA.

MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA biological function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and specificity, and applicability to disease diagnosis and therapy.

Liquid biopsy of circulating tumor DNA and biosensor applications.

Circulating tumor DNA (ctDNA) as a class of liquid biopsy is a type of gene fragment that contains tumor-specific gene changes in body fluids such as human peripheral blood. More and more evidences show that ctDNA is an excellent tumor biomarker for diagnosis, prognosis, tumor heterogeneity and so on. ctDNA is a tumor code in the blood. Liquid biopsy of ctDNA is firstly summarized. Compared with the traditional detection technologies of ctDNA, the biosensor is an excellent choice for the detection of ctDNA because of its portability, sensitivity, specificity and ease of use. This review mainly evaluates various biosensors applied to the detection of ctDNA. We discuss the most commonly used bioreceptors to specifically identify and bind ctDNA, including complementary DNA (cDNA), peptide nucleic acid (PNA) and anti-5 MethylCytosines, and the biotransducers which convert biological signals to analysable signs. The review also discusses signal amplification strategies in biosensors to detect ctDNA.

Recent advances in signal amplification strategy based on oligonucleotide and nanomaterials for microRNA detection-a review.

MicroRNAs (MiRNAs) play multiple crucial regulating roles in cell which can regulate one third of protein-coding genes. MiRNAs participate in the developmental and physiological processes of human body, while their aberrant adjustment will be more likely to trigger diseases such as cancers, kidney disease, central nervous system diseases, cardiovascular diseases, diabetes, viral infections and so on. What's worse, for the detection of miRNAs, their small size, high sequence similarity, low abundance and difficult extraction from cells impose great challenges in the analysis. Hence, it's necessary to fabricate accurate and sensitive biosensing platform for miRNAs detection. Up to now, researchers have developed many signal-amplification strategies for miRNAs detection, including hybridization chain reaction, nuclease amplification, rolling circle amplification, catalyzed hairpin assembly amplification and nanomaterials based amplification. These methods are typical, feasible and frequently used. In this review, we retrospect recent advances in signal amplification strategies for detecting miRNAs and point out the pros and cons of them. Furthermore, further prospects and promising developments of the signal-amplification strategies for detecting miRNAs are proposed.

Gene detection: An essential process to precision medicine.

Gene detection plays an important role in public health and consumer finances cause its prominent functions in precision medicine, pathogens detection and species identification. What's more, the announcement of Precision Medicine Initiative would strengthen its status further in many aspects. Therefore, rapid and simplified gene detection techniques are in urgent demand to meet the practical use. During the last decades, gene detection techniques have advanced considerably by integrating various strategies such as microassay, nanotechnologies and electrochemical methods. Herein, we summerized the recent innovations of gene detection methods based on the signal read-out modalities, and highlight those techniques with the potentials to realize Point-of-care (POC) tests. Additionally, gene detection principle, amplification strategies and their applications are also discussed.

Research advances in the detection of miRNA / 药物分析学报

MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA bio-logical function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and spec-ificity, and applicability to disease diagnosis and therapy.