151 results about "Viral nucleic acid" patented technology

The invention belongs to the technical field of medical treatment, and discloses a blood product virus inactivation system based on a synergistic effect of nano-catalysis photochemical inactivation and low-temperature plasma, the system comprises: a nano-photocatalysis inactivation module adopting titanium dioxide TiO2 and graphite phase carbon nitride g-C3N4 nano-materials as photocatalysts; the low-temperature plasma inactivation module adopts a low-temperature plasma generator to generate plasma cloud above the blood product and release active particles; the dynamic fluid stirring module is used for enabling the blood product to uniformly receive photocatalysis and plasma inactivation effects through a microfluid mixing technology so as to ensure the uniformity of virus inactivation; the sensing monitoring and intelligent feedback module is used for monitoring the residual quantity of viral nucleic acid and the activity of protein in the blood in real time by adopting a spectrum sensor and a biochemical sensor. The method can be widely applied to the fields of clinical blood transfusion, blood product manufacturing, blood disease treatment and the like, promotes the progress of a blood safety guarantee technology, reduces the virus infection risk and improves the patient safety.

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 provides a method for detecting a virus inactivation effect by an ELISA (Enzyme-Linked Immunosorbent Assay) method, which comprises the following steps: acquiring a to-be-detected inactivated virus sample, extracting an antigen component from the sample, and detecting an antigen structure change characteristic value through a high-throughput mass spectrometry analysis technology to obtain antigen degradation degree data; obtaining a sample subjected to preliminary inactivation, and detecting the antigen content change trend through an enzyme-linked immunosorbent assay to obtain content change curve data; detecting the residual quantity of viral nucleic acid by a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) technology aiming at the immunogenicity retained sample to obtain nucleic acid degradation degree data; acquiring nucleic acid degradation degree data, classifying the relationship between nucleic acid residues and infectivity by adopting a support vector machine algorithm, and judging that the inactivation effect is completely completed if the classification result shows that the infectivity is lost; according to the antigen content change trend, the immunogenicity index and the nucleic acid degradation degree data, a weighted fusion algorithm is adopted to integrate multi-dimensional detection results, and a comprehensive inactivation effect score is obtained.

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 relates to the technical field of genetic engineering, and particularly provides a multi-linked quality control product for quality control of a novel coronavirus nucleic acid detection laboratory and a preparation method of the multi-linked quality control product. The multi-linked quality control product is a multi-component composite system and is formed by mixing three substances, namely pseudovirus, target gene RNA (Ribonucleic Acid) molecules and internal standard plasmid DNA (Deoxyribose Nucleic Acid). Wherein the pseudovirus is pseudovirus particles of novel coronavirus ORF1ab gene RNA (Ribonucleic Acid) wrapped by a vesicular stomatitis virus vector and is used for monitoring virus nucleic acid extraction; the target gene RNA molecule is a novel coronavirus N gene RNA molecule and can monitor the reverse transcriptase activity; the internal standard plasmid is plasmid DNA of a human housekeeping gene RPP30, and whether inhibition or failure exists in an amplification link is monitored. According to the method, the whole process of nucleic acid extraction, reverse transcription and amplification is synchronously and hierarchically monitored, the monitoring blind area of a traditional single-component quality control product is solved, and the accuracy and efficiency of laboratory detection quality control are remarkably improved.

The invention relates to a nucleic acid detection platform based on an SHERLOCK technology, which is used for diagnosing CAV-2. The SHERLOCK consisting of recombinase polymerase nucleic acid pre-amplification (RPA) and CRISPR-Cas13a is mainly used for specifically recognizing a conserved sequence of CAV-2RNA (Ribonucleic Acid) through crRNA (Complementary Ribonucleic Acid) so as to activate the non-specific'side cutting activity 'of lwCas13a, so that the report RNA is degraded. A real-time fluorescent quantitative PCR (qPCR) system or a lateral flow test strip is used for collecting fluorescent signals generated by cracking of the reporter molecules, and a visual result is displayed. In addition, through HUDSON treatment, RPA-CRISPR can be amplified at room temperature without viral nucleic acid extraction. The detection limit of the method is 1.1 * 10 < 2 > copies / [mu] L, and the method has no cross reaction with other canine infectious viruses. From sample collection to result output, the whole reaction of RPA-CRISPR can be completed within two hours, and the RPA-CRISPR kit is suitable for rapid test of clinical samples and can work well in the field or in primary laboratories. The method can get rid of the limitation of a detection instrument and complex operation, and is suitable for on-site rapid batch detection. The method is preliminarily verified and further optimized and applied.

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 provides a method for detecting an indicator of a non-viral nucleic acid vector composition. The method comprises the following steps: mixing a permeable nucleic acid dye with a sample containing a non-viral nucleic acid vector composition for marking; detecting the mixture by adopting flow type particle detection equipment, and recording the fluorescence intensity and the particle number of the single-particle-level non-viral nucleic acid vector with positive nucleic acid signals, the particle number of the non-viral nucleic acid vector with negative nucleic acid signals, the fluorescence intensity of free nucleic acid, the particle size of the non-viral nucleic acid vector with positive nucleic acid signals and the like; and calculating to obtain indexes such as empty shell rate, nucleic acid copy number distribution, nucleic acid copy number-particle size distribution and the like of the non-viral nucleic acid vector composition.

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 invention provides systems and methods for analyzing viruses by representing viral genetic diversity with a directed acyclic graph (DAG), which allows genetic sequencing technology to detect rare variations and represent otherwise difficult-to-document diversity within a sample. Additionally, a host-specific sequence DAG can be used to effectively segregate viral nucleic acid sequence reads from host sequence reads when a sample from a host is subject to sequencing. Known viral genomes can be represented using a viral reference DAG and the viral sequence reads from the sample can be compared to viral DAG to identify viral species or strains from which the reads were derived. Where the viral sequence reads indicate great genetic diversity in the virus that was infecting the host, those reads can be assembled into a DAG that itself properly represents that diversity.

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.

A method for measuring drug content indexes of a non-viral nucleic acid vector composition, comprising performing labeling by enabling a permeable nucleic acid dye to be in contact with a non-viral nucleic acid vector (for example, an mRNA-LNP). The drug content indexes are calculated by measuring, at a single-particle level, fluorescent light generated by free nucleic acids of a nucleic acid vector, and fluorescent light generated by externally adsorbed nucleic acids and internally entrapped nucleic acids of the non-viral nucleic acid vector. The drug content indexes include a drug loading capacity, entrapment efficiency and the like. The method optionally further comprises: treating a sample with a nuclease before or after performing labeling by using the permeable nucleic acid dye, and when performing treatment with the nuclease, measuring the fluorescence condition of the non-viral nucleic acid vector composition; or enabling the permeable nucleic acid dye to be in contact with the sample, and then measuring the fluorescence condition of the non-viral nucleic acid vector composition.

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 present invention relates to a rapid, safe, and accurate method for preparing samples containing microbial or viral nucleic acids for amplification and for detecting the presence of highly pathogenic viruses such as Ebola. The method involves heating the sample in the presence of reagents including a solvent and a detergent. Reagents and compositions are also provided.

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.

Provided is a coronavirus detection method which is suitable for a coronavirus disease 2019 (COVID-19) detection. The method includes the following steps. A field-effect transistor-based biosensor (BioFET) platform is provided, wherein the BioFET platform includes a BioFET and a sensor card. The sensor card is detachably connected to the BioFET, wherein the sensor card includes a plurality of sensors and each of the plurality of sensors includes a response electrode. A nucleic acid probe specific to a nucleic acid sequence of COVID-19 virus is immobilized on a surface of the response electrode. A test solution is placed on the response electrode of the sensor card. A pulse voltage is applied to the response electrode, and a detection current generated from the sensor card is measured.

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 application belongs to the technical field of RNA rapid detection, and particularly relates to a rapid amplification and lateral chromatographic detection method for virus RNA and application. The crude virus nucleic acid extract is subjected to a single-tube composite enzyme reaction system reverse transcription, restriction endonuclease digestion and one-way isothermal strand displacement amplification, and then a large amount of specific sequence single-stranded DNA with a marker is generated; in the lateral chromatographic process, the single-stranded DNA is captured by a nucleic acid probe fixed on a detection area of a virus RNA detection test paper, at the same time, avidin labeled on a colloidal gold particle is combined with biotin labeled on the fixed single-stranded DNA, and the colloidal gold particle is indirectly fixed to show a color indication result. The method provided by the application is simple to operate, does not need complicated nucleic acid extraction, does not need large equipment in the whole detection process, and the result can be judged by naked eyes, so that on-site rapid detection can be realized, and the method has the advantages of simplicity, rapidness, high sensitivity and low cost.