21 results about "Virus load" patented technology

The invention belongs to the technical field of aquatic pathogen detection, and provides a primer and probe combination with strong specificity and high sensitivity, a kit and a cdPCR detection method in order to solve the problems of dependence of absolute quantification of an existing qPCR technology on a standard curve and low repeatability and stability caused by uncontrollable quality of a standard substance. The primer probe can generate specific amplification on LYCIV and has no cross reaction on other common aquatic pathogens (such as NNV, DIV1 and the like), so that the specificity of a detection result is fundamentally ensured, a false positive result caused by the cross reaction is effectively avoided, and the diagnosis accuracy is improved. The method has the characteristics of extremely high sensitivity and absolute quantification, the lower limit of detection reaches 6.2 copies / mu L, the method has more advantages in quantification of samples with extremely low concentration, early diagnosis and detection of extremely low virus load can be realized, viruses with extremely low content in fish bodies can be detected earlier, precious time is provided for disease early warning and early intervention, and the method is worthy of popularization and application. Disease outbreak is effectively prevented.

The application discloses a single-tube quantitative detection HBV viral load real-time fluorescent quantitative PCR kit and belongs to the technical field of molecular biological detection. The kit comprises a primer probe combination for single-tube quantitative detection HBV viral load real-time fluorescent quantitative PCR, wherein the primer probe combination comprises a primer pair and a probe for detecting a target gene, and a probe IC-P1 and a probe IC-P2 for detecting an internal standard. The kit can cover various genotypes of HBV viruses, and the minimum detectable viral load is 2.5 IU / mL, the detection sensitivity is high, the repeatability is good, and the quantitative standard curve is constructed without additional quantitative reference products, so that the operation is simplified and the utilization rate of the reagent is improved.

The invention provides qRT-PCR (quantitative real-time polymerase chain reaction) primers, a kit and a detection method for detecting tomato brown wrinkled fruit virus with high sensitivity, and belongs to the technical field of plant virus detection. The qRT-PCR primer for detecting the tomato brown wrinkled fruit virus, disclosed by the invention, is as shown in SEQ ID NO. 1 and SEQ ID NO. 2. The invention also provides a qRT-PCR detection method of the tomato brown wrinkled fruit virus, the primer and the detection method provided by the invention have the characteristics of strong specificity and high sensitivity, the detection limit is 1.562 * 10 < 1 > copies / [mu] L, and the primer and the detection method have a better detection effect on a sample with low virus loading amount; and a technical support is provided for detecting the tomato brown wrinkled fruit virus and preventing and controlling the tomato brown wrinkled fruit virus.

The utility model provides a kind of virus antigen rapid detection test paper, it is related to biological detection technical field, multi-dimensional gradient compound synergistic component, integrate dendritic flow guide groove and nano microporous structure on sample pad, it can promote processing liquid and sample to be radially evenly distributed in diffusion process, on the one hand, effectively avoid the detection interference caused by local liquid concentration too high, on the other hand, significantly shorten the liquid infiltration time of detection starting stage, improve chromatography efficiency, set up gradient distribution's antigen line on NC membrane, its virus specificity antigen concentration is increasing distribution from the end close to binding pad to bibulous pad, accurately adapt the detection requirement of different virus load sample, low virus load sample because of less combined compound, can flow to bibulous pad end high concentration antigen area sufficient combination and produce strong signal, avoid missed detection, high virus load sample because of compound sufficiency, preferentially and binding pad end low concentration antigen area quickly combine, effectively avoid signal inhibition or false negative problem.

The application discloses a single-tube quantitative detection HIV-1 virus load real-time fluorescent quantitative PCR kit and relates to the technical field of molecular biology detection. The kit comprises a primer and probe combination; the primer and probe combination comprises a primer pair GAG-1-F / R and a probe GAG-1-P for detecting a first target gene, a primer pair POL-1-F / R and a probe POL-1-P for detecting a second target gene, and probes IC-P1 and IC-P2 for detecting an internal standard. The kit covers various genotypes of HIV-1 viruses, can stably detect a virus load of 15 IU / mL, has high detection sensitivity and good repeatability, does not need additional quantitative reference products to construct a quantitative standard curve, can simplify operations, and improves the utilization rate of reagents.

The application discloses a kind of based on water body virus load and fish body tissue virus load correlation grouper nervous necrosis virus nondestructive testing method.The method is through optimizing the concentration condition of virus in aquaculture water, realize the efficient concentration and quantification of water body virus load, then combine water-water aquaculture animal virus load correlation model, indirectly and nondestructively assess the virus infection state of aquaculture animal.The application detects water environment instead of detecting animals itself, realizes nondestructive, dynamic, early warning monitoring of the health status of aquaculture animal population, with the advantages of low cost, simple operation, frequent implementation, etc., and has important application value in aquatic disease prevention and control, good seed selection and biological safety management.

Systems for detecting aerosolized viral loads in an airspace include a first and, optionally, a second volume of a liquid. The first volume includes particles sampled from the airspace, and the second volume is free of virus particles. A first conductivity probe is immersed in the first liquid, and a second conductivity probe is immersed in the second liquid. An alternating voltage is applied to each of the first and second conductivity probes. The difference between the resulting alternating currents through the first and second conductivity probes is determined, and is used to estimate the viral load in the first volume, and in the airspace.

The application belongs to the technical field of biology, and particularly relates to a 3D protein mutant O foot-and-mouth disease recombinant virus strain and construction and application thereof. The application mutates methionine at the 403th position of the 3D protein of the O foot-and-mouth disease virus into alanine, and successfully constructs and rescues a 3D protein mutant O foot-and-mouth disease recombinant virus strain rFMDV-M403A by using reverse genetic technology. The amino acid of the recombinant virus strain can be stably mutated in the process of subculture, and the growth speed of the mutant strain is slower than that of the wild strain, and the virus titer is significantly reduced. In individual experiments, the infection of the mutant strain to suckling mice shows that the mutant strain can improve the survival rate of the suckling mice, and significantly reduce the viral load in the liver and the tissue lesions. The application provides an attenuated foot-and-mouth disease recombinant virus strain, which has important significance for in-depth understanding of the pathogenic mechanism and prevention and control strategy of the foot-and-mouth disease virus, and provides an important theoretical basis for further development of the foot-and-mouth disease genetic engineering vaccine.

The invention discloses a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) method for detecting capsicum vein mottle virus in tobacco, which comprises the following steps of: firstly, designing and synthesizing a specific primer pair based on a coat protein CP gene conserved sequence of capsicum vein mottle virus isolate in an NCBI (National Center of Biotechnology Information) database; secondly, extracting total RNA of a tobacco sample infected with the pepper vein mottle virus, and performing reverse transcription to obtain cDNA; carrying out real-time fluorescent quantitative PCR (Polymerase Chain Reaction) amplification by using the obtained cDNA as a template and adopting the specific primer pair; and finally, judging whether the sample carries the target virus or not according to the amplification curve, the melting curve and the Ct value, and realizing quantitative analysis of the virus load through the standard curve. The method is high in specificity and sensitivity, compared with a conventional PCR method, the sensitivity is improved by 10 times, and rapid and accurate detection of the pepper vein mottle virus in the tobacco can be achieved.

The invention belongs to the technical field of pathogen molecular diagnosis and high-throughput sequencing, and particularly relates to a novel bunyavirus whole genome capturing and sequencing method and kit for low virus load and high host background samples. Through the collaborative design of host nucleic acid reduction and two rounds of capture and enrichment, under the condition that the proportion of reads of an initial host source is greater than 90%, a high-coverage sequencing result of high-proportion base sites in L / M / S three fragments of the novel bunyavirus can still be obtained at a relatively low sequencing depth, the coverage integrity and the sequencing sensitivity of a whole genome are remarkably improved, and the method is suitable for large-scale popularization and application. And the sequencing cost and the result volatility are effectively reduced. Reliable whole genome data support can be provided for molecular typing, reassortment / recombination identification, aggregation epidemic situation analysis and transmission chain tracing of the novel bunyavirus, and the method has important clinical and public health application value.

The invention belongs to the technical field of aquatic immunity and biological preparations, and discloses a CXCL20a protein mutant and application of the CXCL20a protein mutant as a grass carp reovirus inactivated vaccine adjuvant. In order to improve the immunomodulatory activity of CXCL20a, the applicant constructs and obtains a CXCL20a mutant protein. After the mutant and a GCRV-II type inactivated vaccine are combined to immunize grass carp, the expression of immune related genes can be remarkably promoted, the activity of serum acid phosphatase and superoxide dismutase is enhanced, and meanwhile, the level of lactic dehydrogenase is reduced. A challenge experiment proves that the survival rate of the grass carp combined with the inactivated vaccine for immunization is increased by about 8% compared with that of a non-mutant combined group, and the in-vivo virus load is reduced. As an immunologic adjuvant, the CXCL20a mutant M-C20 provided by the invention can effectively enhance early immune response and overall protection effect in vaccine immunity, and has good safety, stability and application prospect.

The invention discloses a triple RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) method and a kit for simultaneously detecting IMNV (Immunodeficiency Virus), CMNV (Cytomegalovirus) and DIV1 (Deoxyribose Immunodeficiency Virus According to the invention, PCR primers and TaqMan probes for detecting IMNV, CMNV and DIV1 are respectively designed, and the nucleotide sequences of the PCR primers and TaqMan probes are shown as SEQ ID NO: 1-9. The primer and the probe can be used for simultaneously qualitatively and quantitatively detecting the infectious muscle necrosis virus, the secretly death nodavirus and the full-eye iridovirus 1 through a triple RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) method. According to the invention, common detection of RNA virus and DNA virus is realized for the first time, the kit has the characteristics of high efficiency and convenience, and the advantages of high sensitivity, high specificity and quantitability, and meanwhile, due to the addition of the molecular crowding agent, the detection efficiency and detection speed of medium and low-concentration virus load can be effectively improved, and the detection cost is reduced. The invention provides a convenient and reliable detection method and product for early monitoring, early warning and diagnosis of the three virus diseases of the penaeus vannamei boone.

The invention relates to the technical field of single cell transcriptomics and microbiomics, in particular to a method for detecting viral load based on single cell transcriptome sequencing, which comprises the following steps: extracting single cell transcriptome sequencing data to form a single cell gene expression matrix, and comparing the single cell gene expression matrix with a host comparison file to extract reading data; dynamically translating the reading data to generate an amino acid sequence; comparing and scoring the amino acid sequence with a virus protein sequence in a virus database, obtaining a weighted comparison value through distance weight correction, and forming a virus sequence-cell matrix by the weighted comparison value and reading data; inputting the virus reading data, the host cell immune response index and the cell type sensitivity score into a cell infection tendency index calculation formula, and obtaining the cell infection probability through a logistic regression model. The technical problem that the sensitivity to RNA viruses with high mutation rate is poor is solved, quantitative estimation of the single cell infection probability is achieved, and the method is suitable for detecting distant viruses or new viruses which are not recorded yet.

The application discloses a different genotype goose astrovirus multiplex fluorescent PCR detection primer probe set, a kit and application, relates to the field of molecular biology; the primer probe set comprises a primer pair and a probe for detecting type 1 goose astrovirus (GoAstV-1) and type 2 goose astrovirus (GoAstV-2); the primer pair for detecting GoAstV-1 is as shown in SEQ ID NO. 1-2, and the probe is as shown in SEQ ID NO. 3; the primer pair for detecting GoAstV-2 is as shown in SEQ ID NO. 4-5, and the probe is as shown in SEQ ID NO. 6. The primer probe set disclosed by the application can realize multiplex fluorescent PCR detection of different genotypes of goose astrovirus, the detection method has good specificity, sensitivity and repeatability, and provides technical support for clinical diagnosis of GoAstV, determination of virus load and epidemiological investigation.

The invention discloses a method for preventing and controlling prodenia litura nuclear polyhedrosis virus diseases, and belongs to the technical field of large-scale breeding of insects. The method aims at solving the problems that in the prior art, the chemical disinfection effect is limited, and the disease resistance of prodenia litura is difficult to improve. According to the technical scheme, the method mainly comprises the following steps: firstly, disinfecting prodenia litura eggs to reduce the initial virus load; secondly, an immunopotentiator is added into the basal feed, and the immunopotentiator is selected from sodium houttuyfonate, houttuynia cordata powder or a boiling soak solution prepared from houttuynia cordata roots, stems and leaves, so that the immunologic function of the prodenia litura is enhanced, and the resistance of the prodenia litura to nuclear polyhedrosis virus (NPV) is improved. Through verification, the survival rate of prodenia litura infected with NPV in large-scale feeding can be remarkably increased, and the method is easy and convenient to operate, low in cost and environmentally friendly.

A virus filter includes a filter member characterized by a first surface and a second surface and having a thickness extending between the first surface and the second surface in a first direction, and a plurality of channels formed in the filter member, each of the channels having a channel axis, wherein during use, a solution carrying a viral load flows in a direction parallel to the first surface, and at least a portion of the viral load enters a membrane through the first surface and propagates in the first direction, and wherein for at least 50% of the channels in the filter member, the channel axis is oriented at an angle between 5 degrees and 85 degrees relative to the first direction.

The application relates to a primer probe set, a kit and a method for genotyping of a measles virus B3 / D8 / H1. The method for detecting the measles virus by using the primer probe set and the kit has good sensitivity, the sensitivity of the measles virus of different genotypes can reach 500 copies / mL, and the detection of low virus load can be realized.

The invention discloses a CPA primer group and a kit for detecting seven chicken eimeria coccidiosis and application of the CPA primer group and the kit, and belongs to the technical field of biological detection. Specific primers are designed for conservative COI genes of chicken eimeria, an optimal reaction system and 61 DEG C constant-temperature amplification conditions are optimized and determined, and a CPA detection method for detecting chicken eimeria is established. The method is extremely high in sensitivity, the lower limit of detection on a plasmid standard substance reaches 102 copies / L, extremely low pathogen load equivalent to 0.5 oocyst can be stably detected in practical application, the clinical sample verification coincidence rate reaches 100%, and a powerful technical support is provided for early warning, field monitoring and precise prevention and control of chicken coccidiosis.

This invention provides the application of primer sets in the preparation of reagents for HIV-1 gene drug resistance detection based on next-generation sequencing technology, and a kit thereof. The reagents are used to detect drug resistance mutations in the HIV-1 gene, including subtypes A1, B, C, D, CRF01_AE, CRF07_BC, CRF08_BC, and CRF55_01B. The primer sets generate amplicon lengths of 220-280 bp, and the nucleotide sequences of the primer sets are shown in SEQ ID NO. 1-40. This invention employs a short amplicon enrichment combined with two-round PCR library construction, splicing the full-length pol gene sequence through amplicon overlap regions. Compared to in-house methods based on long fragment amplification, short amplicon methods offer higher amplification stability and detection sensitivity for samples with low viral loads and nucleic acid degradation. Meanwhile, compared to long amplicon fragmentation library construction, this method directly obtains the next-generation sequencing library through two rounds of PCR operations. The experimental procedure is simple and can be completed in just 3.5 hours.

The invention discloses a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) kit for quantitatively detecting HBV (Hepatitis B Virus) load through a single tube, and belongs to the technical field of molecular biological detection. The kit comprises a real-time fluorescent quantitative PCR primer probe combination for quantitatively detecting the HBV virus load in a single tube, and the primer probe combination comprises a primer pair and a probe which are used for detecting a target gene, and a probe IC-P1 and a probe IC-P2 which are used for detecting an internal standard. The kit can cover various genotypes of HBV viruses, can detect the virus load of 2.5 IU / mL to the minimum, is high in detection sensitivity and good in repeatability, does not need additional quantitative reference products to construct a quantitative standard curve, simplifies operation and improves the utilization rate of reagents.

The invention discloses a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) kit for quantitatively detecting HIV-1 (Human Immunodeficiency Virus-1) virus load through a single tube, and relates to the technical field of molecular biological detection. The kit comprises a primer and probe combination, the primer and probe combination comprises a primer pair GAG-1-F / R and a probe GAG-1-P which are used for detecting a first target gene, a primer pair POL-1-F / R and a probe POL-1-P which are used for detecting a second target gene, and a probe IC-P1 and a probe IC-P2 which are used for detecting an internal standard. The kit covers various genotypes of the HIV-1 virus, can stably detect 15 IU / mL virus load, is high in detection sensitivity and good in repeatability, does not need additional quantitative reference substances to construct a quantitative standard curve, can simplify operation, and improves the utilization rate of reagents.