DNA Molecular Computation Using the CRISPR-Mediated Reaction and Surface Growth of Gold Nanoparticles.

DNA has emerged as a promising tool to build logic gates for biocomputing. However, prevailing methodologies predominantly rely on hybridization reactions or structural alterations to construct DNA logic gates, which are limited in simplicity and diversity. Herein, we developed simple and smart DNA-based logic gates for biocomputing through the DNA-mediated growth of gold nanomaterials without precise structure design and probe modification. Capitalizing on their excellent plasmonic properties, the surface growth of gold nanomaterials enables distinct wavelength shifts and unique shapes, which are modulated by the composition, length, and concentration of the DNA sequences. Combined with a CRISPR-mediated reaction, we constructed DNA circuits to achieve complicated biocomputing to modulate the surface growth of gold nanomaterials. By implementing logic functions controlled by input-mediated growth of gold nanomaterials, we established YES/NOT, AND/NAND, OR/NOR, XOR, and INHIBIT gates and further constructed cascade logic circuits, parity checker for natural numbers, and gray code encoder, which are promising for DNA biocomputing.

Novel Nonthermal Atmospheric Plasma Irradiation of Titanium Implants Promotes Osteogenic Effect in Osteoporotic Conditions.

Osteoporosis is a metabolic disease characterized by bone density and trabecular bone loss. Bone loss may affect dental implant osseointegration in patients with osteoporosis. To promote implant osseointegration in osteoporotic patients, we further used a nonthermal atmospheric plasma (NTAP) treatment device previously developed by our research group. After the titanium implant (Ti) is placed into the device, the working gas flow and the electrode switches are turned on, and the treatment is completed in 30 s. Previous studies showed that this NTAP device can remove carbon contamination from the implant surface, increase the hydroxyl groups, and improve its wettability to promote osseointegration in normal conditions. In this study, we demonstrated the tremendous osteogenic enhancement effect of NTAP-Ti in osteoporotic conditions in rats for the first time. Compared to Ti, the proliferative potential of osteoporotic bone marrow mesenchymal stem cells on NTAP-Ti increased by 180% at 1 day (P = 0.004), while their osteogenic differentiation increased by 149% at 14 days (P < 0.001). In addition, the results indicated that NTAP-Ti significantly improved osseointegration in osteoporotic rats in vivo. Compared to the Ti, the bone volume fraction (BV/TV) and trabecular number (Tb.N) values of NTAP-Ti in osteoporotic rats, respectively, increased by 18% (P < 0.001) and 25% (P = 0.007) at 6 weeks and the trabecular separation (Tb.Sp) value decreased by 26% (P = 0.02) at 6 weeks. In conclusion, this study proved a novel NTAP irradiation titanium implant that can significantly promote osseointegration in osteoporotic conditions.

Male sexual dysfunction in patients with chronic kidney disease: a cross-sectional study.

Sexual dysfunction is common in males with chronic kidney disease (CKD), but yet the prevalence and specific relationship between CKD and sexual dysfunction, especially premature ejaculation (PE), remain to be investigated in China; This study aims to examine the prevalence and association between CKD and sexual dysfunction in male patients in China; In this cross-sectional, non-interventional, observational study conducted at a single center. 72 male patients with CKD were enrolled. Data collection included socio-demographic information, assessments via the 5-item version of the International Index of Erectile Function (IIEF-5), the Chinese version of the Premature Ejaculation Diagnostic Tool, the Patient Health Quentionnnaire-9 and the General Anxiety Disorder-7. Data analysis was performed using R version 3.5.2 and SPSS software version 25.0; Among the 72 CKD patients, 56.9% experienced erectile dysfunction and 29.2% had PE. Various factors including estimated Glomerular Filtration Rate, Albumin-to-Creatinine Ratio, psychological aspects, medication use were found to be associated with sexual dysfunction in these CKD patients; Sexual dysfunction is prevalent in males with CKD and is, influenced by multiple factors. It is important for clinicians to focus on sexual dysfunction in this patient group and further investigate its underlying mechanisms.

Small object detection algorithm incorporating swin transformer for tea buds.

Accurate identification of small tea buds is a key technology for tea harvesting robots, which directly affects tea quality and yield. However, due to the complexity of the tea plantation environment and the diversity of tea buds, accurate identification remains an enormous challenge. Current methods based on traditional image processing and machine learning fail to effectively extract subtle features and morphology of small tea buds, resulting in low accuracy and robustness. To achieve accurate identification, this paper proposes a small object detection algorithm called STF-YOLO (Small Target Detection with Swin Transformer and Focused YOLO), which integrates the Swin Transformer module and the YOLOv8 network to improve the detection ability of small objects. The Swin Transformer module extracts visual features based on a self-attention mechanism, which captures global and local context information of small objects to enhance feature representation. The YOLOv8 network is an object detector based on deep convolutional neural networks, offering high speed and precision. Based on the YOLOv8 network, modules including Focus and Depthwise Convolution are introduced to reduce computation and parameters, increase receptive field and feature channels, and improve feature fusion and transmission. Additionally, the Wise Intersection over Union loss is utilized to optimize the network. Experiments conducted on a self-created dataset of tea buds demonstrate that the STF-YOLO model achieves outstanding results, with an accuracy of 91.5% and a mean Average Precision of 89.4%. These results are significantly better than other detectors. Results show that, compared to mainstream algorithms (YOLOv8, YOLOv7, YOLOv5, and YOLOx), the model improves accuracy and F1 score by 5-20.22 percentage points and 0.03-0.13, respectively, proving its effectiveness in enhancing small object detection performance. This research provides technical means for the accurate identification of small tea buds in complex environments and offers insights into small object detection. Future research can further optimize model structures and parameters for more scenarios and tasks, as well as explore data augmentation and model fusion methods to improve generalization ability and robustness.

Mn2+-Mediated Modulation of PfAgo Activity for Biosensing.

Argonaute (Ago) as a powerful enzyme has provided new insights into biosensing due to its programmability, high sensitivity, and user-friendly operation. However, current strategies mainly rely on phosphorylated guide DNA to modulate the cleavage activity of Ago, which is limited in versatility and simplicity. Herein, the authors report the Mn2+-enhanced cleavage activity of Ago and employ Mn-ions with variable valence to regulate the activity of Pyrococcus furiosus Ago (PfAgo) for biosensing applications. The conversion of Mn ions with different valence states through MnO2 nanoflowers enables the sensitive detection of ascorbic acid, alkaline phosphatase, and arsenic with limits of detection of 2.5 nmol L-1, 0.009 U L-1, and 0.4 ng mL-1, respectively. A PfAgo-based immunoassay is further developed that allows for the detection of diverse targets, thus providing a promising toolbox to broaden PfAgo-based sensors into versatile bioanalytical and biomedical applications.

Hierarchical Composite Scaffold with Deferoxamine Delivery System to Promote Bone Regeneration via Optimizing Angiogenesis.

A bone defect refers to the loss of bone tissue caused by trauma or lesions. Bone defects result in high morbidity and deformity rates worldwide. Autologous bone grafting has been widely applied in clinics as the gold standard of treatment; however, it has limitations. Hence, bone tissue engineering (BTE) has been proposed and developed as a novel therapeutic strategy for treating bone defects. Rapid and effective vascularization is essential for bone regeneration. In this study, a hierarchical composite scaffold with deferoxamine (DFO) delivery system, DFO@GMs-pDA/PCL-HNTs (DGPN), was developed, focusing on vascularized bone regeneration. The hierarchical structure of DGPN imitates the microstructure of natural bone and interacts with the local extracellular matrix (ECM), facilitating cell adhesion and proliferation. The addition of 1 wt% of halloysite nanotubes (HNTs) improved the material properties. Hydrophilic and functional groups conferred by polydopamine (pDA) modifications strengthened the scaffold bioactivity. Gelatin microspheres (GMs) protected the pharmacological activity of DFO, achieving local application and sustained release for seven days. DFO effectively promoted angiogenesis by activating the signaling pathway of hypoxia inducible factor-1 α (HIF-1 α). In addition, DFO synergized with HNTs to promote osteogenic differentiation and matrix mineralization. These results indicated that DGPN promoted bone regeneration and accelerated cranial defect healing. This article is protected by copyright. All rights reserved.

The Potential Value of Probiotics after Dental Implant Placement.

Dental implantation is currently the optimal solution for tooth loss. However, the health and stability of dental implants have emerged as global public health concerns. Dental implant placement, healing of the surgical site, osseointegration, stability of bone tissues, and prevention of peri-implant diseases are challenges faced in achieving the long-term health and stability of implants. These have been ongoing concerns in the field of oral implantation. Probiotics, as beneficial microorganisms, play a significant role in the body by inhibiting pathogens, promoting bone tissue homeostasis, and facilitating tissue regeneration, modulating immune-inflammatory levels. This review explores the potential of probiotics in addressing post-implantation challenges. We summarize the existing research regarding the importance of probiotics in managing dental implant health and advocate for further research into their potential applications.

A multitask model for realtime fish detection and segmentation based on YOLOv5.

The accuracy of fish farming and real-time monitoring are essential to the development of "intelligent" fish farming. Although the existing instance segmentation networks (such as Maskrcnn) can detect and segment the fish, most of them are not effective in real-time monitoring. In order to improve the accuracy of fish image segmentation and promote the accurate and intelligent development of fish farming industry, this article uses YOLOv5 as the backbone network and object detection branch, combined with semantic segmentation head for real-time fish detection and segmentation. The experiments show that the object detection precision can reach 95.4% and the semantic segmentation accuracy can reach 98.5% with the algorithm structure proposed in this article, based on the golden crucian carp dataset, and 116.6 FPS can be achieved on RTX3060. On the publicly available dataset PASCAL VOC 2007, the object detection precision is 73.8%, the semantic segmentation accuracy is 84.3%, and the speed is up to 120 FPS on RTX3060.

Gold Nanomaterials-Implemented CRISPR-Cas Systems for Biosensing.

Due to their superiority in the simple design and precise targeting, clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have attracted significant interest for biosensing. On the one hand, CRISPR-Cas systems have the capacity to precisely recognize and cleave specific DNA and RNA sequences. On the other hand, CRISPR-Cas systems such as orthologs of Cas9, Cas12, and Cas13 exhibit cis-cleavage or trans-cleavage activities after recognizing the target sequence. Owing to the cleavage activities, CRISPR-Cas systems can be designed for biosensing by degrading tagged nucleic acids to produce detectable signals. To meet the requirements of point-of-care detection and versatile signal readouts, gold nanomaterials with excellent properties such as high extinction coefficients, easy surface functionalization, and biocompatibility are implemented in CRISPR-Cas-based biosensors. In combination with gold nanomaterials such as gold nanoparticles, gold nanorods, and gold nanostars, great efforts are devoted to fabricating CRISPR-Cas-based biosensors for the detection of diverse targets. This review focuses on the current advances in gold nanomaterials-implemented CRISPR-Cas-based biosensors, particularly the working mechanism and the performance of these biosensors. CRISPR-Cas systems, including CRISPR-Cas9, CRISPR-Cas12a, and CRISPR-Cas13a are discussed and highlighted. Meanwhile, prospects and challenges are also discussed in the design of biosensing strategies based on gold nanomaterials and CRISPR-Cas systems.

Nanotechnology-based diagnostic methods for coronavirus: From nucleic acid extraction to amplification.

The recent emergence of human coronaviruses (CoVs) causing severe acute respiratory syndrome (SARS) is posing a great threat to global public health. Therefore, the rapid and accurate identification of pathogenic viruses plays a vital role in selecting appropriate treatments, saving people's lives and preventing epidemics. Nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are natural biopolymers composed of nucleotides that store, transmit, and express genetic information. Applications of nucleic acid detection range from genotyping and genetic prognostics, to expression profiling and detection of infectious disease. The nucleic acid detection for infectious diseases is widely used, as evidenced by the widespread use of COVID-19 tests for the containment of the pandemic. Nanotechnology influences all medical disciplines and has been considered as an essential tool for novel diagnostics, nanotherapeutics, vaccines, medical imaging, and the utilization of biomaterials for regenerative medicine. In this review, the recent advances in the development of nanotechnology-based diagnostic methods for coronavirus, and their applications in nucleic acid detection are discussed in detail. The techniques for the amplification of nucleic acids are summarized, as well as the use of magnetic nanoparticles for nucleic acid extraction. Besides, current challenges and future prospects are proposed, along with the great potential of nanotechnology for the effective diagnosis of coronavirus.

Construction of a portable immunosensor for the sensitive detection of carbendazim in agricultural products using a personal glucose meter.

Portable and sensitive detection of carbendazim (CBD) is highly desirable for food safety and environmental protection. Herein, a portable immunosensor for the sensitive detection of CBD is proposed based on alkaline phosphatase (ALP)-labeled and secondary antibody-modified gold nanoparticles (AuNPs). The quantification is based on ALP catalyzing the dephosphorylation of glucose-1-phosphate disodium salt to generate glucose, thus converting the concentration of CBD into glucose, thereby realizing the portable detection of CBD by personal glucose meter. Benefiting from signal amplification strategy that integrates the large specific surface area of AuNPs, the enzymatic reactions of terminal deoxynucleotidyl transferase and ALP, a low detection limit of 0.37 ng/mL for CBD is achieved. When this portable method is used to analyze citrus fruit, canned citrus, and cabbage, good-consistency results are obtained with the UPLC-MS/MS method. The good performance demonstrates the great potential of this portable method for CBD monitoring in resource-poor settings.

Comparison of early osseointegration of non-thermal atmospheric plasma-functionalized/ SLActive titanium implant surfaces in beagle dogs.

Background: Hydrophilic dental implants are gaining increasing interest for their ability to accelerate bone formation. However, commercially available hydrophilic implants, such as SLActive™, have some major limitations due to their time-dependent biological aging and lower cost-effectiveness. The non-thermal atmospheric plasma (NTAP) treatment is a reliable way to gain a hydrophilic surface and enhance osseointegration. However, a few studies have been carried out to compare the osseointegration of NTAP-functionalized titanium implants and commercially available hydrophilic implants. Purpose: In this study, we compare the osseointegration abilities of the NTAP-functionalized titanium implant and Straumann SLActive. Material and methods: The NTAP effectiveness was examined using in vitro cell experiments. Then, six beagle dogs were included in the in vivo experiment. Straumann SLActive implants, SLA implants, and SLA implants treated with NTAP were implanted in the mandibular premolar area of dogs. After 2 w, 4 w, and 8 w, the animals were sacrificed and specimens were collected. Radiographic and histological analyses were used to measure osseointegration. Results: NTAP treatment accelerated the initial attachment and differentiation of MC3T3-E1 cells. In the in vivo experiment, bone parameters (e.g., BIC value and BV/TV) and volume of new bone of NTAP groups were close to those of the SLActive group. Additionally, although there was no statistical difference, the osseointegration of SLActive and NTAP groups was evidently superior to that of the SLA group. Conclusion: NTAP-functionalized implants enhanced cell interaction with material and subsequent bone formation. The osseointegration of the NTAP-functionalized implant was comparable to that of the SLActive implant at the early osseointegration stage.

Seed-mediated in situ growth of photothermal reagent gold nanostars: Mechanism study and preliminary assay application.

Photothermal reagent-mediated portable detection platforms using thermometers as signal readers have received extensive attention due to their simplicity, low cost, and practicality. However, exploitation photothermal reagent with excellent photothermal conversion effect, convenient to synthesize, preferably without any modification for biosensing application, is still challenging. Herein, a simple and rapid seed-mediated in situ synthesis strategy has been developed for the preparation of gold nanostars (AuNSs) with remarkable photothermal conversion effect. By simply changing the seed size and component concentrations involved in the in situ synthesis process, AuNSs have adjustable geometries, allowing the photothermal conversion to be tuned to a high level optimal for biosensing. Meanwhile, an accurate understanding of the photothermal conversion mechanism is obtained by studying the relationship between the morphology of AuNSs and the photothermal effect. Subsequently, using ascorbic acid (AA) as a model target, the preliminary application of AuNSs in constructing a portable photothermal detection platform has been demonstrated. This in situ preparation strategy of AuNSs not only exhibits remarkable photothermal conversion effect, but also avoids complicated and time-consuming synthesis and modification. Therefore, it has great potential to be extended to portable detection of other targets by simply converting the concentration of the target to that of AA.

A CRISPR/Cas12a-based photothermal platform for the portable detection of citrus-associated Alternaria genes using a thermometer.

The outbreak of citrus brown spot because of Alternaria is one of the most destructive citrus diseases. Additionally, Alternaria species produce highly toxic mycotoxins. Mass screening is a valid method to control the spread of Alternaria. Morphological analysis and polymerase chain reaction combined with gene-sequencing technique are the most commonly used techniques for detecting Alternaria. However, they are limited by either low convenience and accuracy or low instrument accessibility and high cost. To balance the convenience, accuracy, test availability, and low cost, we develop a CRISPR/Cas12a-based photothermal platform for the portable detection of Alternaria genes using a thermometer. Using this platform, the Alternaria genes from the synthetic sequences and cultured fungus of citrus, tomato, and apple can be detected using a thermometer with a detection limit of 1.5 pM. With the aid of the CRISPR/Cas12a system, citrus-associated Alternaria can be specifically differentiated from other citrus disease-associated microorganisms. When the photothermal platform is applied to analyze the citrus fruit samples collected in the field, good-consistency results are obtained with the gene-sequencing technology. The excellent performance of this portable method shows that it can be applied to screen for Alternaria in resource-poor settings.

Involvement of PI3K/Akt signaling pathway in promoting osteogenesis on titanium implant surfaces modified with novel non-thermal atmospheric plasma.

Non-thermal atmospheric plasma (NTAP) modification to induce a hydrophilic titanium (Ti) surface with less carbon contamination, has been demonstrated to boost the osteogenic responses. In this study, we investigated the underlying bone formation mechanism of NTAP-Ti, and the involvement of PI3K/Akt signaling pathway in regulating osteogenic activities on NTAP-Ti surfaces. NTAP was employed for Ti activation, and PI3K inhibitor, LY294002, was applied to the suppression of PI3K/Akt pathway. We systematically and quantitatively detected the cell morphology, attachment, proliferation, osteogenic differentiation and mineralization of MC3T3-E1 mouse preosteoblasts, and molecular expressions involved in osteogenesis and PI3K/Akt signaling pathway in vivo and in vitro. A descent in osteoblast proliferation on Ti surfaces in relation to LY294002. Alkaline phosphatase (ALP) activity, as well as matrix mineralization, was mitigated by PI3K inhibitor in NTAP-Ti. Likewise, the expression levels of osteogenesis-related genes [ALP, osteocalcin (Ocn), osteopontin (Opn) and runt-related transcription factor 2 (Runx2)] on NTAP-Ti were notably attenuated by LY294002, as confirmed by the results of osteogenesis-related proteins (ALP, and Runx2) expression analysis. In addition, the expression of PI3K/Akt signal pathway proteins further verified the inhibition of LY294002 on Ti surfaces modified by NTAP. Collectively, the PI3K/Akt signal pathway was involved in the amelioration of osteogenesis induced by NTAP modification. NTAP treatment for Ti activation is promising in augmented osteogenic potential through the activation of PI3K/Akt signal pathway.

CRISPR-Cas12a based fluorescence assay for organophosphorus pesticides in agricultural products.

Herein, we propose a sensitive fluorescent assay for organophosphorus pesticides (OPs) detection based on a novel strategy of activating the CRISPR-Cas12a system. Specifically, acetylcholinesterase (AChE) hydrolyzes acetylthiocholine into thiocholine (TCh). Subsequently, TCh induces the degradation of MnO2 nanosheets and generates sufficient Mn2+ ions to activate the Mn2+-dependent DNAzyme. Then, as the catalytic product of activated DNAzyme, the short DNA strand activates the CRISPR-Cas12a system to cleave the fluorophore-quencher-labeled DNA reporter (FQ) probe effectively; thus, increasing the fluorescence intensity (FI) in the solution. However, in the presence of OPs, the activity of AChE is suppressed, resulting in a decrease in FI. Under optimized conditions, the limits of detection for paraoxon, dichlorvos, and demeton were 270, 406, and 218 pg/mL, respectively. Benefiting from the outstanding MnO2 nanosheets properties and three rounds of enzymatic signal amplification, the proposed fluorescence assay holds great potential for the detection of OPs in agricultural products.

The application of DNA-HRP functionalized AuNP probes in colorimetric detection of citrus-associated Alternaria genes.

The monitoring of the fungal genus Alternaria, which causes destructive brown spot disease in citruses worldwide and produces highly toxic mycotoxins, is extremely important to protect citrus and human health. In this work, we describe an ultrasensitive colorimetric method for the detection of genomic DNA of Alternaria from citrus fruit samples, using a system consisting of five groups of reporter probes. Each reporter probe is prepared by coupling recognition DNA and horseradish peroxidase (HRP) on the surface of gold nanoparticle (AuNP) through a convenient and low-cost freezing-assisted method. Meanwhile, the capture DNA is immobilized on magnetic bead (MB) via biotin-streptavidin reaction. Then, the capture DNA, target DNA, and five groups of AuNP-based reporter probes form a stable DNA-heptamer sandwich structure on the MB, and then HRP generates a blue signal for the subsequent colorimetric detection. It should be noted that AuNP with a large specific surface area drives abundant HRP anchoring, resulting in significant signal amplification. In addition, there are five groups of AuNP-based reporter probes, which further amplify the detection signal. As a result, the detection limit of the artificial target DNA is as low as 15.6 pM. Because the detection signal can be recorded visually without any special equipment, and its sensitivity is high, this method represents a suitable diagnostic tool for Alternaria genetic detection.

Transcriptomic profile of human erythroleukemia cells in response to Sargassum fusiforme polysaccharide and its structure analysis.

Sargassum fusiforme (S. fusiforme) has been used as an ingredient in Chinese herbal medicine for thousands of years. However, there are a limited number of studies concerning its therapeutic mechanism. High performance gel permeation chromatography (HPGPC) analysis showed that the average molecular weight of the S. fusiforme polysaccharide, SFPS 191212, is 43 kDa. SFPS 191212 is composed of mannose, rhamnose, galactose, xylose, glucose, and fucose (at a molar ratio: 2.1 : 2.9 : 1.8 : 15.5 : 4.6 : 62.5) with α- and ß-configurations. The present research evaluated the anti-tumor potential of the S. fusiforme polysaccharide in human erythroleukemia (HEL) cells in vitro. To explore the SFPS 191212's apoptosis mechanism in HEL cells, transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212. The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed. It was found that SFPS 191212 inhibited HEL cell proliferation, reduced cell viability in a concentration-dependent manner, and induced an insignificant toxic effect on normal human embryonic lung (MRC-5) cells. Compared with the control group, transcriptome analysis identified a total of 598 differentially expressed genes (DEGs), including 243 up-regulated genes and 355 down-regulated genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on all DEGs, and 900 GO terms and 52 pathways were found to be significantly enriched. Finally, 23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction (qRT-PCR). Moreover, SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway. Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S. fusiforme.

A multicolor enzyme-linked immunoassay method for visual readout of carbendazim.

Enzyme-linked immunosorbent assay (ELISA) with high specificity and sensitivity is one of the most popular techniques for detecting carbendazim (CBD), a commonly used benzimidazole fungicide in agriculture. However, the traditional ELISA based on the horseradish peroxidase (HRP)-3,3',5,5'-tetramethylbenzidine (TMB) system for CBD only displays the yellow color of TMB2+ from deep to light, making it difficult for the naked eye to judge whether CBD in fruits and vegetables exceeds the maximum residue limit. In this article, we intend to improve the traditional ELISA method to establish a multicolor signal output ELISA to achieve visual semiquantitative detection of CBD. This method is based on the optical properties of gold nanorods (AuNRs). After introducing AuNRs into TMB2+ solution, which was produced by the HRP-TMB system of traditional ELISA, AuNRs were quickly etched by TMB2+. Consequently, the longitudinal localized surface plasmon resonance peak of AuNRs shows a clear blue shift and a vivid color change. Different concentrations of CBD generate different amounts of TMB2+, which in turn leads to different etching degrees of AuNRs, and ultimately results in a rainbow-like color change. As a result, CBD from 0.08 to 100 ng mL-1 can be easily distinguished by the naked eye, which does not require any large instruments. Moreover, the colors displayed by 0.49 ng mL-1 (purple) and 0 ng mL-1 (pink) are significantly different from each other. It should be noted that 0.49 ng mL-1 is far below the most stringent maximum residue limit of CBD in the world. Additionally, the quantitative determination of CBD spiked in canned citrus, citrus fruits, chives, and cabbage samples showed satisfactory recoveries. The good performance of the AuNR-based ELISA makes it have a wide range of application prospects in food safety and international trade.

Integrating multiple hybridization chain reactions on gold nanoparticle and alkaline phosphatase-mediated in situ growth of gold nanobipyramids: An ultrasensitive and high color resolution colorimetric method to detect the mecA gene of Staphylococcus aureus.

Colorimetry has been considered as a potential instrument-free platform for point-of-care genomic detection. However, it is limited by the poor sensitivity and low color resolution. Herein, we report a high-resolution colorimetric biosensor based on multiple hybridization chain reactions (HCRs) on gold nanoparticle (AuNP) and alkaline phosphatase (ALP)-mediated in situ growth of gold nanobipyramids (AuNBPs) for ultrasensitive detection of the Staphylococcus aureus (S. aureus) mecA gene. In our design, target DNA is hybridized with capture hairpin DNA on magnetic beads and then amplified by multiple HCRs on AuNP. Since biotin-labeled hairpin-structured nucleic acids are utilized to conduct HCRs, together with the large specific surface area of AuNP, the biotin- and streptavidin- based reaction results in a large amount of ALP on AuNP. With the aid of NADPH, ALP-mediated in situ growth of AuNBPs is observed, and a series of rainbow-like colors are associated with different target DNA concentrations. Through the multiple-amplification strategy produced by AuNP, HCRs, and enzymatic reactions, the target DNA as low as 2.71 pM can be detected with high specificity. Moreover, this method has been successfully applied to detect the mecA gene extracted from S. aureus. Therefore, the proposed method holds great potential in clinical diagnosis.