RESUMEN
The development of sensitive and efficient analytical methods for multiple biomarkers is crucial for cancer screening at early stage. MicroRNAs (miRNAs) are a kind of biomarkers with diagnostic potential for cancer. However, the ultrasensitive and logical analysis of multiple miRNAs with simple operation still faces some challenges. Herein, a photonic crystal (PC)-enhanced fluorescence biosensor with logic gate operation based on one-pot cascade amplification DNA circuit was developed for enzyme-free and ultrasensitive analysis of two cancer-related miRNAs. The fluorescence biosensor was performed by biochemical recognition amplification module (BCRAM) and physical enhancement module (PEM) to achieve logical and sensitive detection. In the BCRAM, one-pot cascade amplification circuit consisted of the upstream parallel entropy-driven circuit (EDC) and the downstream shared catalytic hairpin assembly (CHA). The input of target miRNA would trigger each corresponding EDC, and the parallel EDCs released the same R strand for triggering subsequent CHA; thus, the OR logic gate was obtained with minimization of design and operation. In the PEM, photonic crystal (PC) array was prepared easily for specifically enhancing the fluorescence output from BCRAM by the optical modulation capabilities; meanwhile, the high-throughput signal readout was achieved by microplate analyzer. Benefiting from the integrated advantages of two modules, the proposed biosensor achieved ultrasensitive detection of two miRNAs with easy logic gate operation, obtaining the LODs of 8.6 fM and 6.7 fM under isothermal and enzyme-free conditions. Hence, the biosensor has the advantages of high sensitivity, easy operation, multiplex and high-throughput analysis, showing great potential for cancer screening at early stage.
Asunto(s)
Técnicas Biosensibles , MicroARNs , Técnicas de Amplificación de Ácido Nucleico , Técnicas Biosensibles/métodos , MicroARNs/análisis , Humanos , Técnicas de Amplificación de Ácido Nucleico/métodos , Fluorescencia , ADN/química , ADN/genética , Límite de Detección , Fotones , Lógica , Espectrometría de FluorescenciaRESUMEN
With the emergence of microRNA (miRNA) as a promising biomarker in cancer diagnosis, it is significant to develop multiple analyses of miRNAs. However, it still faces difficulties in ensuring the sensitivity and accuracy during multiplex detection owing to the low abundance and experimental deviation of miRNAs. In this work, a flexible-arranged biomimetic array integrated with parallel entropy-driven circuits (EDCs) was developed for ultrasensitive, multiplex, reliable, and high-throughput detection of miRNAs. The biomimetic array was fabricated by arrangement of various photonic crystals (PCs) for adjustable photonic band gaps (PBGs) and specific fluorescence enhancement. Meanwhile, two cancer-related miRNAs and one reference miRNA were introduced as multiple analytes as a proof-of-concept. The parallel EDCs with negligible crosstalk were designed based on the modular property. Because of the one-to-one match between the emitted fluorescence of parallel EDCs and the PBGs of the flexible-arranged biomimetic array, the generated fluorescence signal triggered by target miRNAs can be enhanced on the corresponding domain of the array. Furthermore, the amplified signal of the array was detected with high-throughput scanning, which could reveal specific information on cancer-related miRNAs as well as reference miRNA, enhancing the abundance and reliability of the analysis. The proposed array has the merits of a modular design, flexible deployment, simple operation (nonenzymatic and isothermal), improved accuracy, high sensitivity, and multiplex analysis, showing potential in disease diagnosis.
Asunto(s)
MicroARNs , Neoplasias , Humanos , MicroARNs/análisis , Entropía , Reproducibilidad de los Resultados , Biomimética , Neoplasias/diagnósticoRESUMEN
Circulating cell-free DNA (cfDNA) is a promising biomarker of liquid biopsy, but it still faces some difficulties in achieving sensitive and convenient detection. Herein, an Ω-shaped fiber optic localized surface plasmon resonance (FO-LSPR) biosensor based on hybridization chain reaction (HCR) coupled with gold nanoparticles (AuNPs) was developed, and applied in simple and sensitive detection of cfDNA. Specifically, one-base mismatch was designed in HCR hairpins (H1 and H2) to obtain high reaction efficiency, and AuNPs was introduced onto H1 through poly-adenine to construct HCR coupled with AuNPs strategy. Meanwhile, target cfDNA was designed into two domains: one could trigger HCR to generate dsDNA concatemer carrying numerous AuNPs, and the other could hybridize with capture DNA on the surface of Ω-shaped fiber optic (FO) probes. Thus, the presence of target cfDNA would initiate HCR, and bring the formed dsDNA concatemer and AuNPs to approach the probe surface, resulting in dramatically amplified LSPR signal. Besides, HCR required simple isothermal and enzyme-free condition, and Ω-shaped FO probe with high refractive index sensitivity just needed to be immersed into HCR solution directly for signal monitoring. Benefiting from the synergetic amplification of mismatched HCR and AuNPs, the proposed biosensor exhibited high sensitivity with a limit of detection of 14.0 pM, and therefore could provide a potential strategy for biomedical analysis and disease diagnosis.
Asunto(s)
Técnicas Biosensibles , Ácidos Nucleicos Libres de Células , Nanopartículas del Metal , Oro , Hibridación de Ácido Nucleico/métodos , ADN/genética , ADN/análisis , Límite de DetecciónRESUMEN
Catalytic hairpin assembly (CHA) appears to be a particularly appealing nucleic acid circuit because of its powerful amplification capability, simple protocols, and enzyme-free and isothermal conditions, and can combine with various signal output modes for the biosensing of various analytes. Especially in the last five years, vast CHA related studies have sprung up. With the deep exploration of the CHA mechanism, some novel and excellent CHA strategies have been proposed; meanwhile the CHA cascade strategies with various amplification techniques further improve the analysis performance. Furthermore, diverse CHA based biosensors have been tactfully engineered and extensively employed in imaging applications in living cells and in vivo ascribed to its gentle reaction, efficient amplification and universality. Hence, we present a comprehensive and systematic summary of the progress in CHA and its application in bioimaging and biomedicine to date. At first, we introduced the mechanism and diversification of CHA in detail, including the newly developed CHA and its ingenious combination with a variety of other technologies. Concurrently, we summarized the latest application progress of different CHA strategies in bioimaging and biomedicine, highlighting the merits and drawbacks of representative approaches. Finally, we put forward some views on the challenges and prospects of CHA in bioimaging and biomedicine in the future.
Asunto(s)
Técnicas Biosensibles , Técnicas Biosensibles/métodos , Catálisis , Hibridación de Ácido NucleicoRESUMEN
Fragrance allergens (FAs) refer to these volatile or semi-volatile fragrance compounds that can cause irritation and negative reactions. A large number of emerging FAs are widely used in household goods, and cause contact allergy or allergic contact dermatitis in eczema population and the general population. It shows an increasing prevalence and is regarded as a concern to public health. Recently, more and more studies have focused on the analytical methods of FAs in a variety of samples with different matrixes. Therefore, a systematic and comprehensive overview of recent progress of analysis of FAs in various samples is needed. In this review, the physical and chemical properties, applications, hazards, and the recent advances of sample preparation and determination methods of common FAs in personal care products, toys, and water samples are systematically and comprehensively summarized. Meanwhile, this review also discusses the advantages and limitations of different sample pretreatment and detection methods, thus offering a deep-going discussion of the development and future trends in this area.
Asunto(s)
Cosméticos , Perfumes , Alérgenos/análisis , Cosméticos/análisis , Humanos , Odorantes , Perfumes/análisis , Perfumes/química , AguaRESUMEN
The development of a universal, sensitive, and rapid assay platform to achieve detections of heavy metal, nucleic acid and bacteria is of great significance but it also faces a thorny challenge. Herein, a novel and universal array platform was developed by combining photonic crystals (PCs) and DNA nanomachine. The developed array platform integrated the physical and biological signal amplification ability of PCs and DNA nanomachine, resulting in ultrasensitive detections of Hg2+, DNA, and Shigella sonnei with limits of detection (LODs) of 22.1 ppt, 31.6 fM, and 9 CFU/mL, respectively. More importantly, by utilizing a microplate reader as signal output device, the array achieved high-throughput scanning (96 samples/3 min) with only 2 µL loading sample, which is advantageous for the detection of infectious dangerous targets. In addition, the PCs array could be obtained easily and rapidly based on self-assembly of colloidal nanospheres, and the DNA nanomachine was operated with enzyme-free and time-saving features. Benefiting from these merits, the proposed PCs array offered a powerful universal platform for large-scale detection of various analytes in the fields of pollution monitoring, epidemic control, and public health.
Asunto(s)
Técnicas Biosensibles , Mercurio , Ácidos Nucleicos , Bacterias , ADN , Límite de DetecciónRESUMEN
Sensitive and efficient monitoring of food-borne bacteria is of great importance for food safety control. Herein, a novel biosensor for highly sensitive detection of Staphylococcus aureus (S. aureus) was constructed by combining hybridization chain reaction (HCR) and nicking enzyme. Different from the upstream-downstream based circuit, the proposed biosensor integrated HCR circuit and three-way DNA junction nicking enzyme assisted signal amplification (3WJ-NEASA) into a virtuous circle of promotion. In the HCR-mediated 3WJ-NEASA sensing strategy, target DNA of S. aureus initiated the self-assembly between HCR hairpins (H1 and H2), which exposed the gap to capture molecular beacon (MB) and construct the 3WJ structure. Meanwhile, MB increased the stability of HCR nanowires and enhanced the efficiency of the HCR circuit, and thus more 3WJ-NEASA circuits were generated in HCR nanowires. Benefiting from the synergistic amplification coupling HCR and 3WJ-NEASA, this isothermal biosensor can detect as low as 6.7 pM of target DNA in one step within only 30 min. Furthermore, the HCR-mediated 3WJ-NEASA assay has been applied in the detection of S. aureus with a limit of detection (LOD) as low as 1.2 × 101 cfu mL-1, and has exhibited reliable practicability in spiked milk. It is the first time that a DNA biosensor combining HCR and 3WJ-NEASA for dual signal amplification was developed and has been adopted to the sensitive analysis of food-borne bacteria. Additionally, this strategy can serve as a universal platform for monitoring other analytes, and therefore possesses broad application prospects in food safety and environmental monitoring.
Asunto(s)
Técnicas Biosensibles , Staphylococcus aureus , ADN , Desoxirribonucleasa I , Límite de Detección , Técnicas de Amplificación de Ácido Nucleico , Hibridación de Ácido Nucleico , Staphylococcus aureus/genéticaRESUMEN
Basal cell nevus syndrome (BCNS), also known as Gorlin-Goltz syndrome, is a rare autosomal dominant genetic disease. It is thought to be caused by a mutation in the PTCH1 gene, and its incidence is 1/57 000 to 1/256 000. The case of a 7-year-old patient with BCNS and Duchenne muscular dystrophy was reported in this paper.