1604 results about "Nucleic acid detection" patented technology

The invention relates to an HPV16 type E6 oncoprotein monoclonal antibody as well as a preparation method and application thereof. The monoclonal antibody specifically bound with the HPV16E6 recombinant antigen is obtained through screening, the monoclonal antibody is applied to a biotin-avidin amplification ELISA system, the E6 oncoprotein of HPV16 can be specifically detected, the sensitivity is 100 pg / mL, the monoclonal antibody does not intersect with oncoprotein of other high-risk HPV, and the E6 oncoprotein of HPV16 can be specifically detected. High specificity and sensitivity are shown for cervical exfoliated cell sample detection of normal people, CIN3 people and cervical cancer people, and the method can be applied to early screening of cervical cancer in the future; compared with traditional exfoliative cytology examination, the method is faster and more accurate, meanwhile, the risk of false positive and misdiagnosis caused by too high sensitivity of nucleic acid detection can be avoided, and the method has very high application value.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody 4F6 for HPV16 type E7 protein detection and application. The heavy chain variable region CDR sequences and the light chain variable region CDR sequences of the antibody are SEQ ID NO.1-3 and SEQ ID NO.4-6 respectively, and the antibody can be specifically combined with HPV16 type E7 protein without cross reaction. A double-antibody sandwich ELISA detection system constructed based on the antibody is combined with a biotin-avidin amplification technology, the sensitivity reaches 100 pg / mL, and the antibody is suitable for rapid detection of the HPV16 type E7 protein in a cervical exfoliated cell sample. The invention further provides the HPV16 type E7 recombinant protein which is obtained through prokaryotic expression and has immunocompetence, and the HPV16 type E7 recombinant protein is applied to antibody screening and detection. Compared with nucleic acid detection, the method has a lower false positive rate, can be used for early screening of cervical cancer and precancerous lesions, and has a good application prospect.

Provided are a double-cavity micro-fluidic chip having a reaction cavity and a test cavity and used for nucleic acid test, and a test method thereof. A fluid channel is provided between the reaction cavity and the test cavity, and only one exhaust channel is provided. The combination of the fluid channel and the exhaust channel can automatically provide resistance for a sample entering the test cavity from the reaction cavity, thereby achieving a controllable transfer of the sample between the reaction cavity and the test cavity. A gravity micro-fluidic chip and a centrifugal micro-fluidic chip are designed. The gravity micro-fluidic chip has a multi-turning circuitous fluid channel of which the diameter is smaller than that of a sample injection channel, such that under the action of gravity, the sample can flow into a reaction cavity from a sample adding region but cannot flow into the fluid channel, and then under pressurization by a simple pressurization device, the sample can enter a test cavity from the reaction cavity. The centrifugal micro-fluidic chip provides an exhaust channel branched from a fluid channel, and makes the exhaust direction opposite to a rotary centrifugal direction and the position of an exhaust opening higher than that of the fluid channel; therefore, by combining the principle of internal and external balance of the atmospheric pressure and the centrifugal driving performance, the centrifugal micro-fluidic chip effectively controls the flowing of a gas and a liquid in reaction cavities and test cavities, so as to effectively control the amount of the sample entering each cavity, so that multiple nucleic acid tests can be completed simply by means of one or two times of centrifugation. In addition, for a chip which only has a reaction cavity, an exhaust channel is connected to a liquid storage cavity, and a gravity-driving chip and an asymmetric pressure-driving chip are designed, thereby effectively avoiding aerosol pollution.

The invention relates to the field of nucleic acid detection, in particular to a multifunctional dumbbell type nucleic acid probe, a kit, a target detection method and application thereof. The multifunctional dumbbell type nucleic acid probe provided by the invention comprises two or more of the following functional areas: a promoter area used for being combined with RNA polymerase; the target recognition area is used for recognizing and combining a target sequence and opening the dumbbell type nucleic acid probe; the dumbbell type nucleic acid probe can be transcribed and combined with a rolling ring, a specific sequence is subjected to mass transcription, amplification of the specific sequence is achieved, finally, signal amplification is achieved during detection, and the dumbbell type nucleic acid probe has the advantages of being simple in reaction system, low in background signal and high in detection specificity; and the dumbbell type nucleic acid probe can be designed according to different target sequences, so that the controllable detection of different targets can be realized.

Provided are an apolipoprotein E detection reagent and the use thereof. The reagent comprises at least one of an ApoE2 protein detection reagent, an ApoE3 protein detection reagent and an ApoE4 protein detection reagent. One of a capture antibody and a detection antibody used for detecting ApoE2 protein or ApoE4 protein is a specific monoclonal antibody, and the capture antibody and the detection antibody used for detecting ApoE3 are both specific monoclonal antibodies. Genotyping of ApoE and quantitative detection of different genotypes of proteins are performed on the basis of an immunological detection method. The detection method has the advantages of being simple, easy to operate, short in time and cheap. A genotype detection result is highly consistent with a fluorescence PCR result. Genotyping of ApoE (6 types) can be performed, which is used to replace the nucleic acid detection and guide medication, and clarify the correlation of the ApoE4 genotype homozygosity / heterozygosity and the protein concentration of ApoE4 with AD.

The invention provides a nucleic acid composition for respiratory tract pathogen detection and application thereof, and relates to the technical field of biology. The invention provides a nucleic acid detection composition which has high specificity and can be used for detecting various respiratory pathogens. The composition can be used for detecting influenza B virus, influenza A virus H1N1, influenza A virus H3N2, novel coronavirus N gene, novel coronavirus O gene, respiratory syncytial virus, I-type parainfluenza virus, II-type parainfluenza virus, III-type parainfluenza virus, IV-type parainfluenza virus, rhinovirus, adenovirus, haemophilus influenzae, and the like. The kit has 15 pathogens including streptococcus pneumoniae and staphylococcus aureus, the lower detection limit can be as low as 20 copies / reaction, and the kit plays an important role in rapid detection of respiratory tract pathogens.

The invention relates to the technical field of deep-sea microorganism nucleic acid detection, in particular to a long-term experimental platform for deep-sea microorganism in-situ nucleic acid detection and a detection method. According to the invention, a water sample is subjected to tangential flow filtration and concentration through the filtration and sampling module, so that a concentrated microorganism sample is obtained. The method comprises the following steps of: transferring a sample into an amplification detection module under reduced pressure, transferring into a lysis purification module for cell lysis, injecting the cracked sample into a chromatographic column, filtering impurities and intercepting DNA by the chromatographic column, then washing by using an eluent to obtain a high-purity DNA sample, loading a suction head through a transfer washing liquid gun, mixing the high-purity DNA sample with an enzyme solution, adding into the amplification detection module for reaction, and detecting the DNA in the amplification detection module. And carrying out fluorescence labeling on the amplified gene, obtaining an amplification curve by using a fluorescence detector, and detecting the in-situ nucleic acid of the deep-sea microorganisms according to the change of the amplification curve. An automatic cleaning assembly is designed in the filtering and sampling module, and the cracking and purifying module is used for sampling and cleaning a cracking chamber and switching pipelines, so that cross contamination caused by deep sea in-situ multiple detection is prevented.

The invention provides an intracellular mycobacterium tuberculosis in-situ nucleic acid detection method based on CRISPR / Cas and application, and the method comprises the following steps: S1, designing crRNA and a primer pair of a mycobacterium tuberculosis target gene sequence, and synthesizing a single-stranded DNA fluorescent probe; s2, preparing a standard solution containing a target gene sequence, and incubating with a CRISPR / Cas12a system; s3, extracting nucleic acids of different types of bacteria, uniformly mixing the nucleic acids of different types of bacteria with the CRISPR / Cas12a system, and incubating; s4, incubating an attenuated strain of mycobacterium tuberculosis and the DNA fluorescent probe, infecting macrophages, and incubating to obtain infected cells; s5, electrically transferring a CRISPR / Cas12a system into the infected cells in S4 by utilizing an electroporation technology, incubating, and detecting the intensity of a fluorescence signal; and S6, performing quantitative and positioning analysis on the detected fluorescence signal to complete detection, and directly introducing a CRISPR / Cas12a system into the macrophage by adopting an electroporation technology to realize intracellular in-situ detection, thereby avoiding the operation complexity of a nucleic acid extraction link and the loss of detection sensitivity in the prior art.

The invention relates to a temperature calibration method of a nucleic acid amplification analyzer, and relates to the field of instrument calibration, a temperature probe of a standard temperature measuring instrument is placed on the instrument, and point value temperatures of 40 DEG C, 50 DEG C, 60 DEG C, 70 DEG C, 80 DEG C, 90 DEG C and 100 DEG C are set; starting the data acquisition instrument, respectively operating and heating to temperatures of different target temperature values, calculating an average temperature value of each hole site, and calculating an absolute value of a difference value between the average temperature value and the target temperature value; a mycoplasma pneumoniae quality control product inspection solution with a standard concentration is used, a matched mycoplasma pneumoniae nucleic acid detection kit is added and then put into an instrument to detect a nucleic acid sample, a temperature drift compensation module is arranged, and the deviation of circuit change in long-term use is corrected through periodic self-inspection; the method has the advantages that the difference between the target temperature value and the actual detection temperature value caused by various factors is eliminated, the temperature drift of the instrument in the long-time use process is solved, and the temperature control accuracy of the instrument in the long-time use process is improved.

The invention relates to the technical field of biomedicine, in particular to a primer probe combination for broad-spectrum screening of common respiratory pathogens and application of the primer probe combination. Through screening and optimization, the respiratory pathogen nucleic acid detection primer probe combination for broad-spectrum screening of common acute respiratory infection and community-acquired pneumonia is obtained; the primer probe combination covers 18 respiratory tract pathogens, has the advantages of high accuracy, good specificity, strong anti-interference capability, good inclusiveness, high sensitivity, hyperjoint detection and the like, can realize parallel detection of 20 targets in cooperation with totally-closed single-tube PCR amplification, greatly improves early diagnosis of respiratory tract diseases with complex pathogen structures, and has wide application prospects. The method has very important significance on prevention, control and treatment of infectious diseases.

The invention relates to the technical field of human papilloma virus detection, in particular to a primer probe combination, a reagent or kit for human papilloma virus nucleic acid detection and application. The purpose of detecting different types is achieved by adding different specific probes, 18 common human papilloma virus DNAs which are internationally recognized to be closely related to cervical cancer can be detected at a time, and HPV16 and HPV18 types are subjected to typing detection. The application provides 6 universal primers (including 4 target primers and 2 internal standard primers) and 19 specific probes (including 18 target probes and 1 internal standard probe), the 4 target primers can amplify DNA of 18 human papilloma viruses, and the 18 probes are specific probes designed for the 18 human papilloma viruses. And meanwhile, by optimizing the primer probe and the reaction liquid, the purpose of detection is achieved.

The invention discloses a method for detecting dengue virus based on RPA / RAA / ERA / MIRA-CRISPR / Cas12a, which comprises the following steps: extracting nucleic acid to obtain a sample to be detected, putting the extracted sample in a sterile and disinfected sample container, contacting the sample to be detected with a composition, a product combination, a reagent, a kit, a system or a system, determining the signal change after the sample to be detected is contacted with different substances, and determining the detection result of the dengue virus based on RPA / RAA / ERA / MIRA-CRISPR / Cas12a. In the same container, a to-be-detected sample is in contact with a composition, a product combination, a reagent, a kit, a system or a part used for RPA / RAA / ERA / MIRA nucleic acid amplification in the system, and then is in contact with a part used for CRISPR nucleic acid detection, the RPA / RAA / ERA / MIRA and CRISPR / Cas12a detection methods are combined, a rapid, simple, sensitive and specific dengue virus detection method is realized, the detection sensitivity is high, and the detection time is short. The reaction conditions of virus detection are further optimized.

The invention relates to the technical field of pathogen detection, relates to a POCT (point-of-care testing) rapid nucleic acid detection system, and in particular relates to a double-stranded probe-assisted loop-mediated isothermal amplification system for detecting brucella. According to the system, an amplification primer and a detection double-stranded probe are designed aiming at a Brucella specific gene bcsp31, and loop-mediated isothermal amplification (LAMP) and a double-stranded probe (TP) based on foot-mediated strand displacement are integrated to form a TP-LAMP unit of the scheme. The amplification result can be monitored in real time through a PCR luminoscope, and can also be visually identified through a nano biosensor (AuNPs-LFB). The method is rapid, sensitive, specific and easy to use, can solve the technical problems of non-ideal detection speed, specificity and sensitivity of a brucella POCT detection technology in the prior art, can be used as a valuable brucella rapid identification tool, and has ideal application and popularization prospects.

The invention relates to the technical field of biosensing, in particular to a method for synergistically activating a CRISPR / Cas12a system based on a T-type structure and application of the method, the CRISPR / Cas12a system can be activated by utilizing two split DNA activation chains and crRNA to form the T-type structure, the activated CRISPR / Cas12a system has trans-cleavage activity, an FQ-ssDNA probe modified with an FAM fluorophore and a BHQ1 quenching group can be subjected to trans-cleavage, a fluorescence signal is recovered, and the fluorescence signal is recovered. Through reasonable design of sources of activation chains corresponding to different targets, detection targets can be expanded from traditional nucleic acid detection to non-nucleic acid targets such as proteins and metal ions, so that biosensing application of multiple targets is realized, in addition, design based on a T-type CRISPR / Cas12a synergistic activation strategy has great flexibility and universality, and the application prospect is wide. The method can be used together with various amplification strategies and detection platforms, and the sensitivity of the analysis method is effectively improved.

The invention provides an RPA-CRISPR-Cas12a system-based detection reagent for the drug resistance of a chilo suppressalis organophosphorus insecticide, a detection method and application, and belongs to the technical field of molecular detection. The invention provides a detection reagent for chilo suppressalis organophosphorus insecticide resistance, the reagent comprises a specific RPA2 primer pair designed for chilo suppressalis acetylcholin esterase ace-1 gene A314S resistance mutation site and crRNA, a double screening mechanism of RPA amplification and CRISPR-Cas12a is combined, wild type and mutant type genes can be accurately distinguished, and the detection reagent can be used for detecting the insecticide resistance of the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis organophosphorus insecticide resistance to the chilo suppressalis. The rapid, sensitive and visual nucleic acid detection of the chilo suppressalis ace-1 gene A314S resistance mutation site is realized, and a visual detection means which is high in specificity, simple and convenient to operate, economical and efficient is provided for field on-site determination of chilo suppressalis resistance populations.

The invention discloses a liquefying agent, application thereof and a nucleic acid detection system, the liquefying agent comprises strong base, acetylcysteine and 2-(N-morpholine) ethanesulfonic acid; in the liquefying agent, the concentration of the strong base is 0.2 mol / L to 1 mol / L; the concentration of the acetylcysteine is 3 to 300 mmol / L; the concentration of the 2-(N-morpholine) ethanesulfonic acid ranges from 2 mmol / L to 5 mmol / L. Through the synergistic effect of the strong base, the acetylcysteine and the 2-(N-morpholine) ethanesulfonic acid, the liquefaction time can be remarkably shortened, the viscous biological sample treatment efficiency and speed can be improved, and the viscous biological sample treated by the liquefier can be directly used for nucleic acid amplification; and the viscous biological sample treated by the liquefying agent can be stored for a long time, the accuracy and reliability of nucleic acid detection are ensured by long-time stability, and the credibility of a detection result is improved.

The invention discloses application of iron ruthenium nano-enzyme in preparation of nucleic acid detection products, and belongs to the technical field of biological detection and nano-materials. The RIN has stable peroxidase-like (POD) activity, and can catalyze a chromogenic reaction between 3, 3 ', 5, 5'-tetramethyl benzidine (TMB) and hydrogen peroxide (H2O2). The catalytic activity of the nano-enzyme can be selectively inhibited by pyrophosphate (PPi) generated in a nucleic acid amplification reaction, so that visual colorimetric analysis on whether target nucleic acid exists or not is realized. In combination with a polymerase chain reaction (PCR) system, the invention constructs a new nucleic acid detection strategy which does not need fluorescence labeling, is simple and convenient to operate and has high sensitivity. Through verification, the system can be widely applied to rapid on-site screening and visual interpretation of related nucleic acid of drug-resistant pathogens (such as ESKAPE flora) and the like, and has a good application prospect.

The utility model belongs to the technical field of micro-fluidic chips, and particularly provides a digital MIRA real-time diagnosis chip and an analysis device.The digital MIRA real-time diagnosis chip comprises a top glass slide, a micro-pattern layer and a bottom glass slide which are sequentially arranged from top to bottom; a water-soluble film is embedded in a position where the micro-pattern layer is matched with the water filling nozzle; the sample injection port is suitable for sequentially injecting a reaction reagent and sealing oil. Through the design of a chip structure without an outlet, when reaction reagents are introduced from a sample injection port, all the reaction reagents simultaneously, equivalently and uniformly enter each micro-chamber, and the reaction reagents are separated into each micro-chamber by adopting an oil seal mode, so that the chip has the advantage of 100% sample discretization in the digital nucleic acid detection process, and no sample or reagent is lost. Meanwhile, each part of reaction reagent is accurately separated into each micro-chamber and cannot be pushed out of the chip, so that each molecule enters the corresponding micro-chamber, and the detection requirement of a low-concentration template is met.

The invention relates to the technical field of nucleic acid detection, in particular to a method for regulating and controlling CRISPR-Cas protein activity through temperature and application of the method, a CRISPR reaction system contains Cas protein, guide RNA, a nucleic acid reporter probe and a buffer solution, and then the cis-cleavage activity or trans-cleavage activity of the Cas protein is activated or inactivated by regulating and controlling the temperature of the reaction system; the guide RNA includes a guide RNA targeting a target nucleic acid sequence that does not contain a PAM site. Multiple detection can be realized only by using the same type of Cas protein, the design of PAM-free sgRNA or crRNA facilitates the selection of sgRNA or crRNA, meanwhile, the activity of PAM-free sgRNA or crRNA can be regulated and controlled by utilizing temperature rise, and the Cas protein with lower reaction temperature can be inactivated, so that the Cas protein cannot cause interference to the detection of the second stage.

The invention discloses a portable nucleic acid detection micro-fluidic chip, a nucleic acid detection device and a nucleic acid detection method, and belongs to the field of micro-fluidic chips, a detection module comprises a quantitative chamber, a micro-channel, a reaction chamber, a first connecting channel and a color developing module; the reaction chamber comprises an RPA reaction chamber in which an RPA amplification reagent is pre-embedded, a porous filter membrane and a CRISPR reaction chamber in which a CRISPR detection component is pre-embedded, and the RPA reaction chamber, the porous filter membrane and the CRISPR reaction chamber are sequentially positioned at the same position of the three adjacent chip layers; the aperture of the porous filter membrane is larger than the particle size of an RPA amplification product and smaller than the particle size of a core enzyme in a CRISPR detection component; two ends of the micro-channel are respectively communicated with the quantitative chamber and the RPA reaction chamber; two ends of the first connecting channel are respectively communicated with the CRISPR reaction chamber and the color developing module; the flow resistance of the first connecting channel is smaller than that of the micro-channel. According to the invention, the sequential reaction of RPA amplification and CRISPR cleavage can be completed without depending on artificial transfer of amplification products.

The invention relates to the technical field of nucleic acid detection, in particular to a product and a method for detecting bifidobacterium longum subsp. Longum 6-1. Bioinformatics analysis is carried out based on a whole genome sequence of the bifidobacterium longum subspecies longum 6-1, the primer pair and the primer probe combination are obtained through optimization, and the primer pair and the primer probe combination have good specificity and sensitivity in the aspect of detecting the bifidobacterium longum subspecies longum 6-1; the bifidobacterium longum subsp. Longum 6-1 and other strains (including other same subsp. Longum strains, other same strains and other strains) can be quickly and accurately distinguished, and the strain level quantitative detection of the strain in a complex biological sample (such as excrement) can be realized.

The invention relates to the technical field of droplet digital nucleic acid detection and biological materials, in particular to microgel based on acrylic anhydride gelatin and nucleic acid to be detected as well as preparation and application of the microgel. The preparation method comprises the following steps: firstly, dissolving a photoinitiator and acrylic anhydride gelatin, and uniformly mixing to obtain a pre-polymerized solution; uniformly mixing the pre-polymerized solution with a to-be-detected nucleic acid mixed solution to obtain a dispersion phase; oil incompatible with the dispersed phase is used as a continuous phase; building a constant-temperature device, and enabling the dispersed phase to form liquid drops by using a liquid drop microfluidic method; and finally, performing amplification treatment on the liquid drops, and then performing photo-initiation crosslinking to obtain the microgel based on the acrylic anhydride gelatin and the nucleic acid to be detected. The prepared microgel based on the acrylic anhydride gelatin and the nucleic acid to be detected is further applied to detection of target nucleic acid.

The invention discloses application of programmable TnpB nuclease gene editing, and relates to the technical field of gene editing. The TnpB nuclease is any one of (A1), (A2) and (A3): (A1) TnpB nuclease, the amino acid sequence of which is as shown in SEQ ID NO.1; (A2) carrying out substitution, deletion and / or addition of one or more amino acid residues on the amino acid sequence of the TnpB nuclease to obtain a protein which has 80% or more of identity with the TnpB nuclease and has the same biological function with the TnpB nuclease; and (A3) a fusion protein obtained by connecting a protein tag to the N terminal or / and C terminal of (A1) or (A2). The TnpB nuclease provided by the invention has cis-cleavage activity and trans-cleavage activity, and has a wide application prospect in the fields of nucleic acid detection and gene editing.

The invention relates to a microfluidic nucleic acid detection device, and relates to the field of detection. Comprising a bottom plate, a chip inlet and outlet module erected on one side above the bottom plate, a chip table erected on the chip inlet and outlet module and used for bearing a micro-fluidic chip with a liquid bag, a displacement module erected above the chip table, and a crushing module erected on the displacement module and used for squeezing liquid in the liquid bag into the micro-fluidic chip, the liquid transferring module is erected on one side of the crushing module and is used for transferring liquid in the micro-fluidic chip to flow; the pneumatic module is erected above the front end of the bottom plate and is used for controlling a flow channel of the micro-fluidic chip to be opened and closed; the heating module is erected on the other side above the bottom plate and is used for locally heating the micro-fluidic chip and automatically detecting the temperature; and the fluorescent module is used for acquiring a fluorescent signal of the micro-fluidic chip. The micro-fluidic chip detection device has the advantages that different types of micro-fluidic chips can be rapidly detected, and an accurate result can be obtained.

The invention discloses a rapid detection method for fusarium wilt in corn based on an RPA-CRISPR / Cas12b technology, and belongs to the technical field of nucleic acid detection. The invention provides a specific reagent combination, a kit and a detection method in order to solve the problems of high amplicon pollution risk, complicated operation and the like in the existing technologies such as RPA (recombinase polymerase amplification). The reagent combination comprises an RPA (recombinase polymerase amplification) primer pair with sequences as shown in SEQ ID NO.1 and SEQ ID NO.2, and sgRNA (single guide ribonucleic acid) with a sequence as shown in SEQ ID NO.3. The method comprises the following steps: carrying out RPA amplification by taking nucleic acid of a sample to be detected as a template, carrying out mixed incubation on an amplification product, sgRNA, Cas12b protein and fluorescent reporter molecules, and judging a result by detecting a fluorescent signal. The optimized RPA and CRISPR systems are integrated on the centrifugal micro-fluidic chip, so that the full-flow closed tube and automatic detection is realized, the detection can be completed within 30 minutes, the detection sensitivity reaches 1E2 copies / T, the risk of aerosol pollution is greatly reduced, and the centrifugal micro-fluidic chip is suitable for port quarantine and field on-site rapid detection.

The invention discloses a multi-subtype influenza A virus nucleic acid detection kit and a detection method, and relates to the technical field of medical detection.The kit comprises a group of reverse transcription RT-RPA amplification primer pairs used for amplifying influenza A virus M gene target sequences, the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9, and the reverse transcription RT-RPA amplification primer pairs are designed for specific areas of subtypes H1N1, H2N2, H3N2, H5N1 and H7N9; the crRNA library comprises a plurality of crRNA molecules which are respectively complementary with different conserved regions in the M gene target sequence and is used for guiding Cas12a protein to carry out specific recognition; a Cas12a protein reagent; the reporter molecule can be subjected to non-specific cleavage by the Cas12a protein; the enzyme preparation, the buffer solution and the nucleotide substrate are required by RT-RPA reaction. The invention aims to realize synchronous, rapid, high-sensitivity and high-specificity detection of various subtype influenza A viruses.

The invention discloses a method for synthesizing a protein-RNA compound in vivo for nucleic acid detection, and relates to the technical field of biology. The technical key points are as follows: a series of recombinant plasmids capable of realizing the simultaneous expression of CRISPR / Cas effect proteins [including all sgRNA-guided Cas proteins such as Cas9, Cas12 (a, b, g, f and the like), Cas13, Cas14 and the like] and crRNA / tracrRNA (sgRNA) thereof in a series of expression strains of escherichia coli and assembling in the escherichia coli body are constructed, wherein the recombinant plasmids can be used for realizing the simultaneous expression of the CRISPR / Cas effect proteins [including all sgRNA-guided Cas proteins such as Cas9, Cas12 (a, b, g, f and the like); according to the present invention, the His tag is added to the 3'end of the compound gene, such that the compounds can be purified by using the nickel column so as to be used for subsequent nucleic acid detection, and the CRISPR / Cas protein nucleic acid compound assembled in vivo has characteristics of high enzyme activity, high sensitivity and high specificity so as to create good conditions for the CRISPR system to establish the nucleic acid detection platform;

The invention discloses a respiratory infectious disease multi-pathogen nucleic acid detection kit based on a fluorescent PCR detection technology and application of the respiratory infectious disease multi-pathogen nucleic acid detection kit. The invention provides two detection kits aiming at 20 and 6 respiratory tract detection, the two kits can specifically detect respiratory tract pathogens, the detection sensitivity is high, the whole detection system is short in time consumption and simple and convenient in use process, and detection results of 20 pathogenic microorganisms can be reported within 40 min at the shortest; the whole set of detection kit and the detection method are high in sensitivity and good in specificity; the normal-temperature storage and transportation of the reagent can be realized.

The invention discloses a system for detecting viral nucleic acid on the basis of a CRISPR (clustered regularly spaced short palindromic repeat) technology and an FET (field effect transistor) chip and application of the system, and relates to a method for detecting the viral nucleic acid on the basis of clustered CRISPR (clustered regularly spaced short palindromic repeat) and related proteins (Cas) and a field effect transistor (FET). The CRISPR-FET (clustered regularly interspaced short palindromic repeats-field effect transistor) detection system comprises a non-amplified CRISPR system, a reporter RNA (ribonucleic acid) and gold nanoparticles (AuNPs) connection system, a separation system and a field effect transistor chip detection system. The CRISPR-FET detection method provided by the invention has high sensitivity and high specificity on virus nucleic acid detection, and the lower limit of detection reaches a single copy (100 copies / [mu] L).

The invention relates to the technical field of phytopathogen detection, and particularly discloses a primer, a kit and a method for detecting crop root rot fusarium solani and application, the primer comprises a specific RPA primer RPA-F and a specific RPA primer RPA-R; the specific RPA primer RPA-F is as shown in SEQ ID NO. 1, and the specific RPA primer RPA-R is as shown in SEQ ID NO. 2; the kit further comprises a guide RNA primer crRNA, and the guide RNA primer crRNA is as shown in SEQ ID NO. 3. The kit comprises a specific RPA primer RPA-F, a specific RPA primer RPA-R, a Cas12 protein, a T7 transcriptase, a crRNA DNA template, a DNA polymerase and a Cas12 / 13 nucleic acid detection test strip. The kit has the characteristics of good sensitivity, relatively good specificity and enough reliability.