106 results about "Viral nucleic acid" patented technology

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 crossing of biomedical engineering, molecular diagnosis and nano-optical sensing, and particularly discloses a virus nucleic acid variation amplification-free LSPR (Local Surface Plasmon Resonance) detection method and chip based on a gold nano-structure. The virus nucleic acid variation amplification-free LSPR detection chip based on the gold nanostructure comprises a sample injection port, a reaction area, a washing area, a detection area and a waste liquid area, the sample injection port is used for injecting a viral nucleic acid or protein sample, a plurality of groups of probe sequences are fixed in different reaction regions of the chip, non-amplification rapid detection of viral nucleic acid variation is realized, high sensitivity and specificity are ensured, and by utilizing the unique advantages of an MSNs template branched gold nanostructure, the detection sensitivity is improved. Refractive index change caused by single base mutation is directly amplified, the capability of completing femtomole-level detection within 30 minutes is achieved, the overall detection speed is increased, rapid typing detection of viral nucleic acid variation is achieved through LSPR spectral signals under the non-amplification condition, and a portable platform is formed by integrating the LSPR spectral signals and a micro-fluidic chip.

The invention belongs to the technical field of viral nucleic acid molecular diagnosis, and particularly relates to a kit for detecting cucumber green mottle mosaic virus based on enzyme-mediated dual-amplification nucleic acid amplification, a detection method and application. The detection system comprises a primer pair DNA upstream primer (SEQ ID NO.2) and a DNA downstream primer (SEQ ID NO.3) for amplifying a coat protein gene of the cucumber green mottle mosaic virus, and a probe (RNA primer (SEQ ID NO.1)) for detecting a CP gene, a nucleic acid amplification group enzyme system, a signal amplification group enzyme system, a nucleic acid scavenging enzyme, an activating enzyme NTP, a buffer solution and a freeze-drying protective agent, wherein the system is prepared into pre-subpackaged freeze-dried balls. The kit provided by the invention can realize RNA detection of aM concentration, and can realize early screening of cucumber green mottle mosaic virus due to high detection sensitivity.

The invention discloses a construction and verification method of an HEK293T cell line capable of stably overexpressing human ANT2, which comprises the following steps: cloning a human ANT2 gene, constructing the human ANT2 gene into a lentivirus expression plasmid pLVX-TRE3G, co-transfecting the lentivirus expression plasmid pLVX-TRE3G and a helper plasmid pLVX-Tet3G into an HEK293T-ACE2 cell, and carrying out puromycin and G418 pressurized screening and Western blot identification to obtain the HEK293T cell line capable of stably overexpressing human ANT2. A result shows that the expression quantity of the ANT2 is the highest when the HEK293T-ACE2-ANT2 cell is induced for 12 hours at the Doxycycline of 6mg / L; when the ANT2 is induced to express, the virus nucleic acid load and the N protein expression level after SARS-CoV-2 infection are obviously reduced, which indicates that the ANT2 has an inhibition effect on SARS-CoV-2 infection replication, and a cell model and an experimental basis are provided for exploring the effect of the ANT2 in virus infection resistance.

The invention provides a method and a kit for quantitative detection of recombinant redistribution of a Databa virus based on a multiple digital PCR (Polymerase Chain Reaction) technology, and belongs to the field of infectious disease monitoring. The method comprises the following steps: providing a to-be-detected sample containing segmented viruses and viral nucleic acid; detecting nucleic acid by using a plurality of multiple digital PCR reaction systems containing specific primer pairs and fluorescence labeled probes; amplifying according to a fluorescence coding rule, collecting and analyzing multi-channel fluorescence signals, and calculating the absolute copy number of each genotype; and based on copy number analysis genotype intra-fragment distribution proportion and inter-fragment combination mode, evaluating characteristics and frequency of virus recombination reconfiguration events. According to the detection method and the kit provided by the invention, accurate quantification of all main genotypes of the Databa virus can be realized.

The invention provides a tsutsugamushi disease pathogen detection primer, a probe and a kit, the sequences of the primer and the probe in an asymmetric primer ERA detection system are designed and screened according to the target sequence of a tsutsugamushi disease pathogen gene, and the proportion of upstream and downstream primers and an amplification reaction system are optimized; the nucleic acid sequence of the tsutsugamushi disease pathogen can be specifically recognized, and non-specific binding with other irrelevant nucleic acid is reduced, so that the detection specificity is improved; meanwhile, efficient amplification of the tsutsugamushi disease pathogen nucleic acid is achieved, accurate detection can be achieved even under the condition that the virus nucleic acid content is extremely low, and therefore the detection sensitivity and the amplification efficiency are improved.

The application discloses a low-cost hepatitis B (HBV) virus nucleic acid rapid detection method based on a functionally modified paper base, and can realize rapid detection of HBV virus nucleic acid.The method comprises the following steps: (1) pre-amplifying HBV virus nucleic acid to obtain Texas red-labeled nucleic acid; (2) capturing and enriching the Texas red-labeled nucleic acid by using a functionally modified paper base; and (3) performing signal detection.The functionally modified paper base is modified with a fusion protein containing a TR512 polypeptide.The protein functionally modified paper base is simple in preparation, fast in nucleic acid detection time and low in detection cost.

The present application relates to rainbow trout nucleic acid vaccine adjuvant, the adjuvant includes polydisulfide compound and cyclic di-GMP delivered by polydisulfide compound, the present application also relates to the application of the rainbow trout nucleic acid vaccine adjuvant in the preparation of rainbow trout IHNV virus nucleic acid vaccine.A kind of rainbow trout nucleic acid vaccine adjuvant, i.e.c-di-GMP+CPD complex, is developed based on STING agonist-c-di-GMP and polydisulfide compound (CPD) delivery carrier, so as to realize the cost reduction and efficiency increase of rainbow trout nucleic acid vaccine, polydisulfide compound (CPD) can efficiently deliver cyclic di-GMP, and have good stability, c-di-GMP+CPD complex has the potential as rainbow trout immune adjuvant, can significantly enhance the immune protection effect of rainbow trout nucleic acid vaccine, reduces vaccine dosage.

The invention discloses a primer probe composition for detecting measles, parotitis, rubella and varicella virus nucleic acid and application, and belongs to the technical field of virus nucleic acid detection. The invention protects the real-time fluorescent PCR detection primer and probe composition for simultaneously or respectively detecting measles, parotitis, rubella and varicella viruses and different subtype strains, and plays a positive role in respectively or simultaneously performing clinical detection and prevention and diagnosis on different strains of several viruses. The detection method disclosed by the invention is simple, time-saving and labor-saving, wide in application range and convenient to use, and measles, rubella, parotitis and varicella viruses and different strains can be rapidly detected at the same time by amplifying a PCR reaction once. The primer probe composition provided by the invention is high in detection sensitivity, strong in specificity and good in repeatability, can realize rapid differential diagnosis on clinical samples, enables different patients to obtain timely targeted prevention and treatment, and has important significance for development of quadruple vaccines.

The invention discloses a kit for identifying a G2b gene subtype of a porcine epidemic diarrhea virus and application of the kit, and belongs to the field of virus nucleic acid detection. Through gene comparison, primers and probes are designed at specific sequence positions of the PEDV G2b gene subtype, and quantitative PCR identification detection can be directly carried out through an LNA technology. The detection method only comprises two steps of nucleic acid extraction and one-step fluorescent RT-PCR reaction, can complete identification within 2 hours at the soonest, has the characteristics of good specificity, high sensitivity and good stability, can rapidly, simply, conveniently and efficiently identify and detect PEDV G2b subtype nucleic acid, is low in cost and short in period, is especially suitable for high-throughput detection, and can be widely applied to detection of PEDV G2b subtype nucleic acid. And the method has important significance on rapid identification of PEDV G2b gene subtype strains.

The invention relates to the technical field of virus extraction kits, in particular to a large-volume sample virus nucleic acid rapid extraction kit and method.The large-volume sample virus nucleic acid rapid extraction kit comprises a protease K solution, magnetic beads, a lysis solution, a rinsing solution 1, a rinsing solution 2 and an eluent; the magnetic beads are hydroxyl magnetic beads; the lysate is prepared from guanidine isothiocyanate, Tris alkali, Tris hydrochloric acid, triton X-100, isopropyl alcohol and polyoxyethylene ether; the rinsing liquid 1 is composed of a guanidine hydrochloride solution and isopropanol; the rinsing liquid 2 is composed of absolute ethyl alcohol and purified water; the kit provided by the invention can easily obtain viral nucleic acid from a large-volume sample, is simple to operate, can extract a plurality of samples at one time, realizes rapid diagnosis, and makes up the vacancy of a kit for automatically extracting viral nucleic acid from a large-volume sample at high flux in the current market.

The invention discloses a macaque cytomegalovirus detection kit supporting multi-mode interpretation and a detection method, and particularly relates to the technical field of virus detection. The kit comprises a nucleic acid quick release reagent, a primer group, an isothermal amplification reaction system, a CRISPR (clustered regularly interspaced short palindromic repeats) detection system, a visual detection component, positive and negative controls, related buffer solutions, enzymes and the like. The detection method integrates nucleic acid extraction-free, ERA isothermal amplification and CRISPRCas12a technologies, realizes rapid amplification and specific detection of target virus nucleic acid, and supports three result interpretation modes of fluorescence, ultraviolet visualization and a lateral flow chromatography test strip. The method has high sensitivity and specificity, the whole process from sample treatment to result output can be completed within 50 min, complex instruments and nucleic acid purification steps are not needed, operation is easy and convenient, cost is low, and the method is suitable for on-site rapid screening of the macaque cytomegalovirus and basic-level facility application.

The invention provides a multiplex RPA (recombinase polymerase amplification) detection primer probe group, a test strip, a kit and a detection method for three prawn RNA (ribonucleic acid) viruses, and belongs to the technical field of biological detection. The invention provides a multiple RPA (recombinase polymerase amplification) detection primer probe group for three prawn RNA (ribonucleic acid) viruses. The multiple RPA detection primer probe group is designed by taking an IMNV MCP gene, a CMNV RdRp gene and a PvPV RdRp gene as target genes. The primer probe set designed by the invention meets the requirements of multiple RPA detection, and can specifically amplify corresponding viral nucleic acid. The invention also provides multiple RPA (recombinase polymerase amplification) detection capable of completing the three RNA viruses of prawns. When the triple RPA detection method constructed by the invention is used for detecting IMNV and CMNV in a sample, the result is completely consistent with the result of an RT-qPCR detection method. The triple RPA detection method established by the invention can be used for simultaneously detecting three viruses in a shorter time, and has a remarkable on-site detection advantage.

The invention provides a real-time fluorescent quantitative PCR (polymerase chain reaction) detection primer, kit and method for novel variant bocavirus, and belongs to the technical field of virus nucleic acid detection. A specific primer is designed and synthesized according to a conserved sequence of a novel variant bocavirus (BoV) VP gene, a recombinant plasmid pcDNA3.1 HA-BoV-VP is constructed, then the recombinant plasmid is used as a standard substance to establish a standard curve, a real-time fluorescent quantitative PCR detection system for the novel variant bocavirus is constructed, and specific detection of the novel variant bocavirus is realized. The detection method can realize quantitative detection of bocavirus in a to-be-detected sample, has the characteristics of high speed, high sensitivity, strong specificity and the like, plays an important role in protection of endangered wild animals such as disc sheep, and has good practicability.

The application discloses a kind of RNA virus nucleic acid preservative in fecal sample, including denaturant, chelating agent, pH buffering agent, osmotic pressure regulator and reducing agent;The denaturant includes guanidine salt and surfactant;The application further additionally provides a kind of RNA virus nucleic acid preservative in fecal sample preparation method, preparation above-mentioned one kind of RNA virus nucleic acid preservative in fecal sample, including the following steps: each component is mixed with ratio, pH is adjusted with pH buffering agent, with deionized water constant volume;Filter, obtain the preservative solution described in this application.The preservative solution of the application can stably store human cell DNA and viral RNA in fecal sample at room temperature for at least 7 days, which is beneficial for long-distance transportation at room temperature;Meanwhile, the preservative solution of the application also releases nucleic acid, inactivates virus, so as to prevent biological leakage and infection phenomenon;The de-inhibitor component in the preservative solution can directly combine and precipitate with various inhibitors in feces, reducing the interference to nucleic acid detection.

This application relates to the field of biomedical detection technology, specifically to a combination of biomarkers for assessing the severity of viremia in Chikungunya virus patients and its application. The combination of biomarkers for assessing the severity of viremia in Chikungunya virus patients includes one or more of the following biomarkers: TAP1, ISG15, CMPK2, GBP4, OTOF, and SIGLEC1. The biomarker combination provided in this application can objectively and quantitatively reflect the activity of viral replication and the strength of the immune response in the patient, thereby achieving accurate assessment of the severity of viremia. This solves the problem in existing technologies that mainly rely on qualitative or semi-quantitative detection of viral nucleic acids and lack assessment methods closely related to the host's pathological state.

This disclosure concerns methods of analysing and sequencing viral genomic DNA, in particular Epstein-Barr virus (EBV) genomic DNA, using viral-specific nucleic acid capture probes and long read sequencing.

The invention provides a primer probe group and a kit for detecting an Acaspot virus, an Oya virus and a Peaton virus based on triple RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction), and belongs to the technical field of molecular biology and nucleic acid detection. According to the present invention, by designing the primers and the probes for the Acaspot virus S fragment, the Oya virus S fragment and the Peaton virus M fragment, the Ct value is determined through the RT-qPCR method, and the existence of the virus nucleic acid and the virus type are determined according to the determination standard, such that the method has advantages of high sensitivity, good specificity, low detection cost, convenience, rapidness and the like, and is suitable for promotion and application in actual detection.

The invention discloses a pH-responsive isoGPBG hydrogel and an application of the pH-responsive isoGPBG hydrogel in miRNA delivery, and belongs to the technical field of biopesticides. The pH-responsive isoGPBG hydrogel disclosed by the invention is formed by deprotonating isoguanosine and guanosine under an alkaline condition and cross-linking the deprotonated isoguanosine and guanosine with a boric acid group of PBA to form a pH-sensitive phenylboronic acid ester bond. The hydrogel delivery system can break through plant miRNA transmembrane transport obstacles, and efficient delivery and controllable release of antiviral nucleic acid drugs are achieved. When the hydrogel delivery system is used for delivering miRNA, in-vitro degradation of miRNA can be prevented, and the hydrogel delivery system has an important application prospect in prevention and treatment of plant viral diseases and bacterial diseases.

This invention discloses a feline calicivirus nucleic acid detection kit and detection method. The detection kit includes: (1) a nucleic acid enrichment module and (2) a TaqMan fluorescent PCR detection module. The beneficial effects are that the sensitivity is significantly improved, the operation is simplified, the functions are integrated, the pathogenicity is classified in one step (distinguishing between highly pathogenic strains and classic strains), the stability is excellent, and the specificity is strong.

The invention discloses an injector-based nucleic acid purification device, an extraction method and a viral nucleic acid lysate. The purification device comprises an injector assembly and a filter membrane assembly, wherein the injector assembly comprises an injector tube barrel and a push rod matched with the injector tube barrel; the filter membrane assembly is connected to the outlet end of the injector tube barrel, and the filter membrane assembly is configured to be of a detachable structure. The nucleic acid purification device provided by the invention perfects the processes of nucleic acid release, nucleic acid washing and elution, can remove components which interfere with subsequent PCR (Polymerase Chain Reaction), fluorescent quantitative PCR and constant-temperature detection in the nucleic acid release process, and can improve the detection efficiency; the viral nucleic acid lysate is suitable for common clinical samples such as nasal swabs, anus swabs, serum and ascites, and is suitable for nucleic acid extraction and purification of DNA and RNA viruses by optimizing the proportion of guanidine isothiocyanate, polyvinylpyrrolidone (PVP-40) and lauryl sodium sulfate (SDS).

This invention belongs to the field of molecular biology detection technology, specifically relating to a Nipah virus (NiV) nucleic acid detection kit combining RT-LAMP and immunochromatography, and its application. This kit combines the high sensitivity and specificity of loop-mediated isothermal amplification (LAMP) technology with the rapid and visual interpretation advantages of immunochromatographic test strips, enabling on-site, rapid, and accurate detection of NiV. The core components of this invention include: a set of specific primers designed for conserved regions of the NiV gene, an optimized RT-LAMP reaction system, and specially treated immunochromatographic test strips for detecting LAMP amplification products. This kit eliminates the need for reverse transcription of RNA virus nucleic acids and expensive thermal cycling equipment. The entire detection process is completed within 20–30 minutes, with a sensitivity of up to 300 copies / μL. This solution combines the high efficiency of RT-LAMP technology with an intuitive, equipment-free detection method, achieving truly rapid and accurate on-site diagnosis of NiV.

The application belongs to the technical field of virus nucleic acid molecule diagnosis, and particularly relates to a kit for detecting cucumber green mottle mosaic virus based on enzyme-mediated double amplification of nucleic acid, a detection method and application. The detection system of the application comprises a primer pair for amplifying the coat protein gene of the cucumber green mottle mosaic virus, i.e. a DNA upstream primer (SEQ ID NO. 2) and a DNA downstream primer (SEQ ID NO. 3), a probe for detecting the CP gene (an RNA primer (SEQ ID NO. 1)), a nucleic acid amplification enzyme system, a signal amplification enzyme system, a nucleic acid clearing enzyme, an activating enzyme NTP, a buffer, and a freeze-drying protective agent, which are prepared into a pre-dispensed freeze-dried ball. The kit can realize RNA detection at aM concentration, and early screening of the cucumber green mottle mosaic virus can be realized due to high detection sensitivity.

The invention relates to the technical field of gene detection, in particular to omega RNA for detecting bovine nodular skin disease virus, a fluorescence detection kit, a visual kit and application thereof. Based on a detection method combining an isothermal amplification technology and a TnpB-omega RNA gene editing system, a novel kit for detecting bovine nodular skin disease virus nucleic acid is developed. The kit is simple and convenient to operate and low in manufacturing cost, has the detection advantages that the reaction speed is high, the sensitivity is high, the specificity is good, and the result can be observed by naked eyes, can realize rapid and efficient detection of bovine nodular skin disease virus nucleic acid, and has a relatively good application prospect.

The invention relates to the technical field of virus detection, in particular to a primer probe combination, a kit and a method for detecting G3 gene group pig bocavirus based on a constant-temperature RAA fluorescence method in a non-diagnostic purpose mode. Aiming at a conserved region of a PBoV G3 virus genome, a probe which is specially modified and designed is adopted and has a synergistic effect with an upstream primer as shown in SEQ ID NO.4 and a downstream primer as shown in SEQ ID NO.7. The primer probe combination is used for detecting the G3 gene group pig bocavirus by a constant-temperature RAA fluorescence method, the detection sensitivity and specificity can be remarkably improved, viral nucleic acid as low as 10.5 copies / mu L can be detected in an extremely short time, meanwhile, cross reaction with other pig bocavirus gene groups or common pig pathogens is effectively avoided, and the detection sensitivity and specificity are improved. And a dual detection mode of instrument quantification and blue light naked eye interpretation can be supported, so that the detection efficiency and the convenience of field application are effectively improved.

The invention provides an RNA (Ribonucleic Acid) virus nucleic acid enrichment method which comprises the following steps: selecting an enrichment target viridae, and designing an enrichment primer based on the enrichment target viridae to obtain a target primer group; the method comprises the following steps: acquiring a to-be-detected animal tissue sample, extracting total RNA from the to-be-detected animal tissue sample, and performing reverse transcription to obtain sample cDNA; enriching the RNA virus nucleic acid in the sample cDNA by adopting the target primer group to obtain an enriched product; wherein the target primer group comprises upstream primers from the first to the ninety-sixth and downstream primers from the first to the ninety-sixth, the sequences of the upstream primers from the first to the ninety-sixth are as shown in SEQ ID NO.1-SEQ ID NO.96, and the sequences of the downstream primers from the first to the ninety-sixth are as shown in SEQ ID NO.97-SEQ ID NO.192. The primer group disclosed by the invention has the characteristics of low cost and stable performance.

The invention discloses a complete virus particle detection quantification system and method based on droplet microfluidics, and belongs to the field of micro-nano technology, the system comprises a single virus adsorption microsphere and two subsystems; the single virus adsorption microspheres are coupled with an antibody through a streptavidin chemical bond, and single viruses are adsorbed by virtue of antigen-antibody specific binding; the single virus nucleic acid detection subsystem wraps microsphere liquid drops with viruses, water-phase liquid containing microsphere-virus samples and a fluorescent probe reagent are subjected to RT-PCR amplification, single virus nucleic acid is detected in the liquid drops, and fluorescent probes are used for calibrating and comparing fluorescence intensity. And the high-throughput liquid drop screening subsystem analyzes the size of the liquid drop and the intensity of the fluorescence signal, identifies the liquid drop containing the single virus, and accurately counts the fluorescence liquid drops, so that quantitative analysis on the single virus level is realized. Complete virus particles are identified by adopting a method of combining nucleic acid detection and protein detection, and high-sensitivity detection and absolute quantitative analysis on a single virus level are realized by adopting a droplet microfluidic technology.

This invention belongs to the field of rapid detection technology, specifically relating to a rapid, real-time detection method for influenza A based on electrochemical biosensors. This invention provides an aptamer that specifically recognizes the influenza A virus, and uses a pairing affinity reagent to enrich the aptamer onto magnetic beads to prepare a probe that specifically recognizes the influenza A virus. Then, using this probe, MB solution, and screen-printed electrodes, an electrochemical biosensor that specifically recognizes the influenza A virus is fabricated. Qualitative or quantitative detection of influenza A virus using the electrochemical biosensor of this invention eliminates the need for complex pretreatment and viral nucleic acid amplification, greatly simplifying the operation. The results are stable and reliable, making it suitable for real-time detection applications.

The application relates to application of dihydroergotamine mesylate in preparation of anti-influenza virus drugs and belongs to the technical field of medicines. The application provides application of dihydroergotamine mesylate in preparation of anti-influenza virus drugs. The dihydroergotamine mesylate can play a role in an early replication stage after influenza virus enters a host cell, can inhibit synthesis of influenza virus nucleic acid, can significantly improve lung symptoms caused by influenza A, reduces virus titer in lung tissues, has good cell safety, biological safety and effectiveness, and has a good anti-virus application prospect.

The present invention relates to the technical field of virus detection, and discloses a sewage diarrhea virus typing detection system, which comprises a pathogen enrichment unit, an improved polyethylene glycol precipitation method is adopted to carry out pathogen enrichment on a sewage sample; and the nucleic acid extraction unit is used for extracting nucleic acid in the enriched sample by using a magnetic bead extraction method. According to the typing detection system for the diarrhea viruses in the sewage, simultaneous detection and typing of various diarrhea viruses such as norovirus, rotavirus, adenovirus, astrovirus, parareovirus and fiveleaf virus in the sewage can be realized, the detection range is widened, powerful support is provided for comprehensive monitoring of virus pollution conditions in the sewage, and the system is suitable for popularization and application. According to the present invention, the multiple fluorescent RT-PCR technology is adopted, and the efficient sample enrichment and nucleic acid extraction method is combined, such that the detection sensitivity is improved, the low-concentration viral nucleic acid can be detected, and the potential viral threat is not missed;