78 results about "Affitin" patented technology

The application provides a preparation method of a functionalized phage probe and application of the functionalized phage probe in detection of escherichia coli. The functionalized phage is a recombinant phage (M13KO7@SaBP) in which streptavidin binding peptide (SaBP) is fused and expressed at the N terminal of the main capsid protein P8 protein of M13K07 phage. In the application, the gene encoding SaBP (MDVEAWLGAR) is inserted into the N terminal of the P8 protein gene of M13K07 phage by site-directed mutagenesis to prepare M13K07@SaBP. The stable signal amplification is realized by using the abundant P8 protein (about 2700) of M13 phage and the super strong binding force of biotin-streptavidin, the interference of food sample matrix is reduced by combining the magnetic separation technology, and a detection method of escherichia coli with high sensitivity and small sample matrix interference is constructed.

This invention provides a double-antibody sandwich SA-ELISA detection kit for Fasciola hepatica infection in sheep. A double-antibody sandwich SA-ELISA method for detecting Fasciola hepatica CAg (cag) is established using mouse anti-Fasciola hepatica D38 monoclonal antibody and rabbit anti-Fasciola hepatica D38 polyclonal antibody. This detection uses monoclonal antibodies as coating antibodies, resulting in high specificity. Furthermore, the introduction of a biotin-streptavidin signal amplification system enhances sensitivity. It offers advantages such as high accuracy, good stability, simple operation, rapid detection, and easy interpretation, making it suitable for rapid clinical diagnosis and epidemiological surveys at the grassroots level.

This invention discloses a homogeneous microsphere based on singlet oxygen self-circulating aggregation-induced emission, its preparation, and a wash-free detection method, which improves the sensitivity and efficiency of wash-free luminescent oxygen channel immunoassay. This invention successfully prepared AIE receptor microspheres by embedding the photosensitizer AIER dye, ultra-bright AIEG dye, and singlet oxygen-responsive dye SO into carboxyl-modified polystyrene nanoparticles. Simultaneously, an antibody-1-labeled AIE receptor microsphere solution, an antigen-containing test solution, and biotinylated antibody-2 were added to a 96-well plate, followed by streptavidin-modified donor microspheres. After wash-free incubation, the fluorescence signal value was measured, and the concentration of antigen in the test solution was obtained according to a standard curve. This platform completes wash-free, highly sensitive detection within 5 minutes, innovatively combining the aggregation-induced emission characteristics of AIEgens and a singlet oxygen cycling strategy, providing an efficient and universal solution for wash-free diagnostics.

Disclosed herein are chromatographic materials comprising crosslinked streptavidin molecules. The crosslinked streptavidin chromatographic materials may be used to prepare affinity chromatographic materials using biotinylated affinity agents, such as a biotinylated antibody, biotinylated antigen-binding fragment, or biotinylated oligonucleotide. Due to the crosslinked nature of the streptavidin molecules, the materials provided herein have reduced to no streptavidin leachate, resulting in improved performance.

A kind of biotin or avidin labeled fat body and its preparation method, nano-carrier system. Biotin or avidin labeled fat body is prepared using specific amount of Bio-phospholipid or avidin-phospholipid, and the prepared fat body has the advantages of high purity, small particle size, good uniformity and high stability. Based on the biotin-avidin system, a nano-carrier system is constructed, which includes the biotin labeled fat body and the avidin modified target component. The system enables the fat body to carry any interested substance, such as viral recombinant protein, antibody and other protein drugs, nucleic acid drugs or small molecule drugs, etc., so as to be used for the treatment of diseases such as vaccines, cancer, infectious diseases and metabolic diseases.

The invention discloses a biosensor based on an ultraviolet light triggered bio-barcode release technology (OBCA) and construction and application of the biosensor. The biosensor comprises nucleic acid functionalized silver nanoparticles, nucleic acid functionalized magnetic beads and a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas12a detection system, the DNA bar code modified on the surface of the silver nanoparticle contains a light cleavable linker, a CRISPR / Cas12a activation sequence and a target capture sequence I, and a DNA capture probe containing a target capture sequence II is fixed on the surface of the magnetic bead through biotin-streptavidin action. During detection, target miRNA mediates two capture sequences to form a sandwich compound, after magnetic separation and thermal denaturation elution, 365nm ultraviolet radiation releases DNA bar code fragments, and the Cas12a trans-cleavage fluorescence quenching reporter probe is activated to generate a signal. According to the method, nucleic acid amplification and reverse transcription are not needed, the detection limits of miR-21, miR-155 and mi-210 respectively reach 158.94 aM, 763.56 aM and 197.82 aM, single-base mismatch and precursor miRNA can be distinguished, the serum sample detection result is highly consistent with that of RT-PCR, and the method is suitable for high-sensitivity quantitative detection of miRNA.

The application provides a skin interstitial fluid miRNA multicolor visualization semi-quantitative detection method, which comprises the following steps: incubating a skin interstitial fluid sample with magnetic nano probes of surface immobilized hairpin probes H1, opening H1 and triggering a subsequent hybridization chain reaction of hairpin probes H2 and H3 in the presence of target miRNA, and constructing a biotin-rich DNA complex on the surface of the magnetic beads; performing enzyme labeling by using an alkaline phosphatase-streptavidin coupling compound, and catalyzing ascorbic acid phosphate to generate ascorbic acid potassium iodate, so that the color of the solution changes, and high-sensitivity detection of the target miRNA is realized. The multicolor visualization detection method can produce near-full-spectrum color changes by combining the gold nano double-cone local surface plasmon resonance property change, and is beneficial to improving the portability of the sensor and the accuracy of the visual analysis.

This invention relates to an alanine aminotransferase (ALT) detection reagent and its preparation method, belonging to the field of medical testing. The ALT detection reagent comprises nanobodies AC1 and AT1 binding to different ALT binding sites. Nanobodies AC1 are biotinylated and immobilized on an avidin ELISA plate as a capture antibody, while nanobodies AT1 serve as the detection antibody. The ALT detection reagent and method prepared based on these nanobodies can meet the needs for rapid and accurate ALT detection.

The invention relates to ribozyme for labeling biotin on target RNA (Ribonucleic Acid) as well as a screening method and application thereof. The RNA labeling technology is one of core tools for modern molecular biology research and clinical diagnosis. A specific marker is covalently linked to a target RNA molecule, so that accurate tracking, efficient detection and functional analysis of RNA can be realized. A biotin-streptavidin system becomes one of the technical routes which are most widely applied due to extremely high affinity and signal amplification capability of the biotin-streptavidin system. At present, common biotin-labeled RNA technical methods mainly comprise a chemical solid-phase synthesis method, an enzymatic method and a chemical labeling method. However, the methods have the limitations of poor site specificity, low efficiency, complex manipulation and the like in the use process. The engineering ribozyme for catalyzing RNA biotinylation is obtained by taking 12: 0 biotin coenzyme A as a biotin donor through an in-vitro screening technology. The ribozyme provided by the invention can efficiently and specifically carry out biotin labeling on target RNA, the reaction process is simple, and the labeling activity is up to 90% or above.

The invention discloses an engineered exosome preparation with tumor targeting and specific gripper response release functions as well as a preparation method and application of the engineered exosome preparation. The preparation comprises a myocardial cell exosome, an NK cell biotinylation activation antibody, a tumor disease biotinylation antibody, streptavidin, a tumor fiber matrix FAP targeting peptide and a tumor matrix enzyme response peptide, and anchoring is carried out through an acetamide compound-polyethylene glycol anchor point platform. The engineering bispecific exosome preparation with good tumor targeting and barrier breakthrough ability and immune cell activation effect is prepared, and the preparation breaks a solid tumor compact fiber barrier and increases NK cell infiltration by utilizing the fibrosis treatment ability of the myocardial cell exosome and the capture and targeting effects of an anchor point platform; the natural killer cells are in close contact with tumor cells, the lasting tumor cell killing effect is achieved, and the tumor matrix environment is fundamentally changed.

The invention relates to the field of single-molecule gene sequencing, in particular to a composite nucleotide probe based on a nucleic acid nanostructure and application of the composite nucleotide probe. The structure of the composite fluorescent nucleotide probe provided by the invention is a multi-level structure, the multi-level structure respectively comprises a nano nucleic acid molecule, streptavidin and a biotin-nucleoside polyphosphate compound from inside to outside, and the nano nucleic acid molecule, the streptavidin and the biotin-nucleoside polyphosphate compound are connected through biotin. The nucleic acid chain, biotin modification and fluorescent dye molecule modification assembled into the nucleic acid nanostructure can be ordered and synthesized by commercial synthesis companies, the price is low, and synthesis is rapid and convenient; the three-dimensional branched connecting molecules can fully disperse the dendritic macromolecules with large volumes in a three-dimensional space. The planar branched connecting molecule is used for connecting the nucleoside polyphosphate and the polyethylene glycol dispersing element, so that the distance between the nucleoside polyphosphate and the polyethylene glycol is relatively short, the polyethylene glycol can conveniently separate and protect the nucleoside polyphosphate, and the diffusion effect of the nucleoside polyphosphate in an aqueous solution is improved.

The present invention belongs to the technical field of genetic engineering, and specifically relates to a copepod luciferase mutant and the use thereof. The mutant is obtained by means of performing one or more mutations on a wild-type copepod luciferase with the signal peptide removed, and can be expressed in a prokaryotic expression system, resulting in an increased luminescence half-life or a signal rise within a certain time range, and the luminescence pattern of the luciferase is converted to glow-type luminescence or luminescence with a sustained increase in luminescence intensity. Some copepod luciferase mutants exhibit enhanced luminescence intensity. The mutant is widely used in biological research for real-time tracking and continuous monitoring, such as for labeling and tracking activities or processes in living organisms. On the basis of the advantage of increased luminescence half-life of the copepod luciferase mutant, the mutant can be used as and / or in the preparation of a glow-type luciferase. The copepod luciferase mutant is more suitable as a reporter gene for in-vivo detection in scientific research, and is conjugated with avidin, a digoxin antibody, etc., for sequencing, thereby shortening the sequencing time and improving the sequencing quality.

The application discloses a kit for determining anti-sperm antibody IgG and application thereof, and comprises sperm membrane antigen coated carboxyl magnetic beads, an analysis buffer and an enzyme working solution, wherein the sperm membrane antigen coated carboxyl magnetic beads are labeled with biotin, streptavidin and streptavidin antibody; in the application, the streptavidin antibody is added in the coating process of the carboxyl magnetic beads, a carboxyl magnetic bead-streptavidin antibody-streptavidin-biotinized antigen is formed, the connecting length between the antigen and the magnetic beads is increased, the steric hindrance is reduced, and the interference of some sample detection is avoided.

Disclosed herein are chromatographic materials comprising streptavidin molecules, wherein the streptavidin molecules are bound to biotin and one or more biotinylated affinity agents. The biotinylated affinity agent may be a biotinylated antibody, biotinylated antigen-binding fragment, or biotinylated oligonucleotide. The streptavidin chromatographic materials may be used to prepare affinity chromatographic materials. Due to biotin binding to the conjugated streptavidin, the materials provided herein have reduced to no streptavidin leachate, resulting in improved performance.

The invention relates to a preparation method and application of a'split 'bionic preparation based on zombie macrophages, biological targeting and mechanical response are combined to improve the drug delivery efficiency of damaged endothelium in a complex blood flow environment so as to efficiently treat AS, and by preparing the zombie macrophages with freeze shock, the treatment effect of the zombie macrophages is improved, and the treatment effect of the zombie macrophages is improved. The preparation method comprises the following steps: taking zombie macrophages as a template, modifying Anti-CD45-biotin-streptavidin on the surface of the zombie macrophages, and utilizing an affinity reaction between the streptavidin and Anti-CD45-biotin to obtain a split type bionic preparation based on the zombie macrophages; according to the split type bionic preparation, firstly, drug-loaded LNP is precisely delivered to AS focus inflammatory blood vessels through the natural targeting homing capacity of a zombie macrophage carrier which is subjected to freezing shock, the zombie macrophage rolls and adheres to the surface of endothelial cells, so that the drug-loaded LNP effectively makes contact with the damaged endothelial cells, and by means of the force sensitivity characteristic of dsDNA, the drug-loaded LNP and the damaged endothelial cells can be effectively released. Finally, the release of the drug-loaded LNP from the macrophages is realized.

The invention discloses a nano antibody for detecting glycinin, an ELISA kit and application thereof, and belongs to the technical field of glycinin detection. Wherein the nano antibody comprises a nano antibody 1 and a nano antibody 2 which can be respectively used as a coating antibody and a detection antibody (biotin label) in the ELISA kit; the ELISA kit can also comprise reagents such as enzyme-labeled streptavidin, a color developing solution and the like, and the quantitative analysis of the glycinin in a to-be-detected sample is realized by specifically utilizing a sandwich ELISA method and a color developing reaction. According to the scheme, the specificity is high, the detection limit is low, glycinin residues in products such as fermented soybeans, enzymolysis soybeans, soybean meal and feed can be accurately and sensitively detected, meanwhile, the defects that in traditional instrument detection, sample pretreatment is complex, consumed time is long and the like are overcome, and the detection cost is greatly saved.

The invention belongs to the technical field of biology, and particularly relates to a reagent for detecting pathogenic genes as well as a preparation method and application thereof. The invention finds that a biological sample is enriched through broad-spectrum captured protein, the captured protein is labeled with biotin, pathogenic microorganisms are separated from the biological sample through high-density streptavidin coated magnetic beads, the pathogenic microorganisms are split after pure culture, pathogenic genes are detected after nucleic acid is extracted, and the turnover time is shortened to 4 hours. The precise distribution of nucleic acid is realized through an annular array reaction cavity preloaded with 32 freeze-dried LAMP reagents in combination with centrifugal force driving. The single chip synchronously detects more than or equal to 30 major pathogen genes / drug-resistant genes, covers common bloodstream infection pathogens, and breaks through the existing POCT flux bottleneck. According to the present invention, with the kit, the blood matrix inhibition can be eliminated, the pathogen concentration can be rapidly increased (the detection threshold value can be reached after culture), the positive detection rate can be increased to the clinical requirement level (gt, 90%), and the pathogen detection limit in the blood sample can meet the clinical critical infection early diagnosis requirement of less than or equal to 10 CFU / mL;

The application belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting human pregnancy-associated plasma protein A and application thereof. The combination comprises monoclonal antibodies 4H5 and 1G8, and CDR amino acid sequences of heavy chain and light chain variable regions of the antibodies are shown in SEQ ID NO. 1-SEQ ID NO. 12. The combination can be used to construct a double antibody sandwich ELISA detection system, 4H5 is used as a coating antibody, 1G8 is biotin-labeled, signal amplification is realized by combining biotin-avidin, and the detection sensitivity is significantly improved. The method can effectively distinguish serum samples of patients with acute coronary syndrome and healthy people, and is suitable for rapid and sensitive detection of biomarkers related to cardiovascular diseases in primary clinical practice.

The invention relates to a polar aprotic organic solvent for dissociating a streptavidin-biotin conjugate as well as a screening method and application of the polar aprotic organic solvent. Aiming at the problems that a streptavidin-biotin conjugate system is difficult to elute quickly under a mild condition and a carrier is difficult to regenerate, a solvent molecule descriptor database is constructed; a polar aprotic organic solvent which meets specific molar refractive index and polarity parameters and is a non-hydrogen bond donor is screened from a large number of candidate solvents by utilizing a machine learning model. The solvent can realize rapid reversible dissociation of biotin conjugates from streptavidin at room temperature, and the carrier still keeps high activity after multiple rounds of binding-elution cycles. The solvent is further used for purification of biotinylated small molecules in a crude reaction solution, enrichment and elution of mitochondria and cell surface samples and the like, topological resolution proteomics analysis is achieved, a high-purity product can be obtained in a high-flux mode, and site level topological analysis can be completed.

The present disclosure is generally directed to the use of biomaterial scaffolds engineered with SA-FasL for the transplantation of stem cell derived β-cells as a treatment for Type I diabetes. Early engraftment post-transplantation and subsequent maturation of these β-cells may be limited by the initial inflammatory response, which impacts the ability to sustain normoglycemia at long times. The survival and development of immature hPSC-derived β-cells transplanted on poly(lactide-co-glycolide) (PLG) microporous scaffolds into the peritoneal fat, a site being considered for clinical translation, was investigated. The scaffolds were modified with biotin for binding of a streptavidin-FasL (SAFasL) chimeric protein to modulate the local inflammatory microenvironment. The presence of FasL impacted infiltration of monocytes and neutrophils and altered their phenotypic response. Conditioned media generated from scaffolds explanted at day 4 did not impact hPSC-derived β-cell survival and maturation in vitro, which was not observed with unmodified scaffolds. Following transplantation, β-cell viability and differentiation were improved with SA-FasL modification. A sustained increase in insulin positive cell ratio was observed with SA-FasL modified relative to unmodified scaffolds. These results demonstrate that SA-FasL-modified scaffolds can mitigate initial inflammatory response and enhance β-cell engraftment and differentiation.

The invention discloses a method for promoting coupling of DNA oligonucleotides and macromolecules by using ice-phase confinement and application of the method, and belongs to the technical field of biological material preparation. The method comprises the following steps: freezing single-stranded DNA containing a terminal reaction group and a buffer solution of macromolecules to be coupled under the condition of-20 DEG C, and enriching solutes in a micro water sac area between ice crystals, so that local concentration and reaction site accessibility are remarkably improved, and various coupling chemistry (SPAAC, CuAAC, biotin-streptavidin and the like) can be quickly and freely completed by a template at low temperature. The method is high in universality, mild in condition and low in reagent consumption, high-efficiency construction of long-chain DNA, 3 '-3' / 5 '-5' inverted connection and internal site modification can be achieved, the coupling efficiency of antibody-DNA and BSA-DNA can be remarkably improved under the condition that protein conformation and functions are maintained, a valence-controllable nucleic acid-protein composite structure is achieved, and the method is suitable for industrial production. The method is suitable for the fields of nucleic acid probe development, protein engineering, nano material construction, drug delivery and the like.

The invention provides an anti-CD4 antibody as well as a microsphere flow immunofluorescence luminescence detection kit and application thereof, and provides two monoclonal antibodies for recognizing different binding sites of a CD4 antigen, an antibody coupling system based on magnetic fluorescent microspheres, a biotinylation detection antibody and phycoerythrin labeled streptavidin. The invention also discloses a corresponding detection method and a preparation method of the kit. The CD4 antigen detection kit and the antibody combination thereof provided by the invention can significantly improve the sensitivity, stability, specificity and flux capacity of CD4 detection, and are suitable for multiple fields of immune function evaluation, autoimmune disease evaluation, inflammatory disease research, HIV infection auxiliary diagnosis and the like.

The invention discloses a nano antibody for detecting beta-conglycinin, an ELISA kit and application of the nano antibody and the ELISA kit, and belongs to the technical field of beta-conglycinin detection. Wherein the nano antibody comprises a nano antibody 1 and a nano antibody 2 which can be respectively used as a coating antibody and a detection antibody (biotin label) in the ELISA kit; the ELISA kit can also comprise reagents such as enzyme-labeled streptavidin, a color developing solution and the like, and the quantitative analysis of the beta-conglycinin in a to-be-detected sample is realized by specifically utilizing a sandwich ELISA method and a color developing reaction. According to the scheme, the specificity is high, the detection limit is low, beta-conglycinin residues in products such as fermented soybeans, enzymolysis soybeans, soybean meal and feed can be accurately and sensitively detected, meanwhile, the defects that in traditional instrument detection, the sample pretreatment process is tedious, operation is complex and time-consuming are overcome, and the detection cost is greatly saved.

The present application relates to the technical field of protein detection, and particularly relates to a detection method and kit for detecting anti-Nephrin antibody. The detection method comprises the following steps: using biotinylated Nephrin protein to capture anti-Nephrin antibody in a sample to be detected on a SA matrix to obtain a first complex; performing acidolysis desorption, neutralization and Protein G directed antibody immobilization on the first complex to obtain a second complex; using a biotin-streptavidin-HRP system to detect the second complex; the acidolysis desorption comprises the following steps: using an acid solution with a pH of 2-3 to treat the first complex, and treating at 35-40 DEG C and 500-800 rpm for 10-20 minutes. The method provided by the present application can accurately and quickly detect the anti-Nephrin antibody level in a sample, and has important application value.

Disclosed herein are antigen chromatography columns. Said antigen chromatography columns utilize streptavidin-functionalized particles, wherein the streptavidin binding sites are occupied with one or more biotinylated antigens. In this way, said antigen chromatography columns may be used to isolate, purify, and characterize antibodies (or other proteins) that specifically bind to said antigens.

The invention relates to a chemiluminiscence detection kit for a blood marker pTau217 of an Alzheimer's disease and application of the chemiluminiscence detection kit. The kit comprises a magnetic microsphere suspension coated with a pTau217 capture antibody; a pTau217 detection antibody labeled by PEG12 (Polyethylene Glycol) < 12 > biotin; streptavidin marked by acridinium ester; an interference eliminating liquid; a washing liquid; pre-excitation liquid and excitation liquid. A PEG12-biotin-streptavidin system is introduced, a large amount of acridinium ester is coupled through streptavidin, a total luminescence signal is amplified, and the detection sensitivity and precision of the chemiluminescence pTau217 are greatly improved. Meanwhile, interference elimination liquid containing dioxane and Proclin300 is adopted and mixed with a plasma sample, then antibody capturing microspheres are added, non-specific adsorption of lipid and binding protein thereof in plasma on the surfaces of the microspheres is weakened, and the detection accuracy is greatly improved.

The invention provides an anti-ADGRG1 antibody as well as a microsphere flow type immunofluorescence luminescence detection kit and application thereof, and provides two monoclonal antibodies for recognizing different binding sites of an ADGRG1 antigen, an antibody coupling system based on magnetic fluorescent microspheres, a biotinylation detection antibody and phycoerythrin labeled streptavidin. The invention also discloses a corresponding detection method and a preparation method of the kit. The ADGRG1 antigen detection kit and the antibody combination thereof provided by the invention can significantly improve the sensitivity, stability, specificity and flux capacity of ADGRG1 detection, and are suitable for multiple fields of immune function evaluation, autoimmune disease evaluation, inflammatory disease research, HIV infection auxiliary diagnosis and the like.

The application relates to a protein sample processing method, comprising the following steps: mixing a protein sample with an alkylating agent to seal free cysteine, obtaining a sealed sample; centrifuging the sealed sample through an ultrafiltration membrane, obtaining a membrane sample; the pore size of the ultrafiltration membrane ranges from 10 kDa to 30 kDa; performing first reduction on the membrane sample, obtaining a first reduced sample; adding a biotin labeling agent to the first reduced sample, obtaining a labeled sample; enzymatically digesting the labeled sample, obtaining an enzymatically digested sample; adding avidin microbead agarose to the enzymatically digested sample to adsorb biotin-labeled proteins, obtaining an enriched sample; performing reduction and alkylization on the enriched sample, obtaining a processed sample.

The invention provides a full-automatic immunodetection method for amplifying a signal, which comprises the following steps: controlling a pipetting device to transfer a sample to be detected, a reagent 1 containing a solid phase carrier coated with a first ligand, a reagent 2 containing a second ligand labeled with biotin and a reagent 3 containing streptavidin labeled with a signal substance into a reaction container; a second compound is formed, and the second compound is a solid phase carrier-first ligand-object to be detected-second ligand-biotin-streptavidin-signal substance compound; the first ligand and the second ligand are specifically combined with different structural domains of an object to be detected; in the second compound, the molar ratio of the biotin to the second ligand is greater than or equal to 2: 1, the molar ratio of the signal substance to the streptavidin is greater than or equal to 2: 1, and the molar ratio of the streptavidin to the second ligand is greater than or equal to 1: 1. According to the detection method, cascade amplification of detection signals is achieved, the detection sensitivity is greatly improved, and accurate detection of the to-be-detected substance with extremely low concentration is achieved.

This invention discloses a freeze-dried RAA-CRISPR / Cas13a one-pot system and its application for on-site detection of Mycobacterium tuberculosis. The freeze-dried RAA-CRISPR / Cas13a one-pot system includes Cas13a protein and crRNA, or a complex of the two, and a lateral flow test strip for use with it. The crRNA includes an anchoring sequence for binding to the Cas13a protein and a guide sequence for targeting the Mycobacterium tuberculosis target sequence. The Mycobacterium tuberculosis target sequence is shown in SEQ ID No. 1. The lateral flow test strip sequentially includes a sample plate, a gold nanoparticle anti-FITC antibody, a streptavidin line, an antibody capture line, and an absorption plate. The antibody coated on the antibody capture line is goat anti-rabbit IgG. This invention has good sensitivity and specificity, and a simple and rapid detection of Mycobacterium tuberculosis nucleic acid can be achieved by a single-step reaction at room temperature. Moreover, the reaction process does not require opening the cap, solving the problem of aerosol contamination that is prone to occur in two-step detection methods.