109 results about "Absolute quantification" patented technology

The invention relates to the technical field of biological detection, relates to a multiple ddPCR detection method for porcine intestinal coronavirus, and in particular relates to a monitoring and quantitative detection method for samples with relatively low porcine intestinal coronavirus content or complex matrix, such as feeds, animal tissues and animal-derived products. The droplet digital PCR detection method for the porcine intestinal coronavirus established in the invention has the advantages of strong specificity, high sensitivity, good repeatability and strong anti-interference capability, can be used for absolute quantification of the copy concentration of the porcine acute diarrhea syndrome coronavirus (SADS-CoV), the porcine epidemic diarrhea virus (PEDV), the porcine delta coronavirus (PDCoV) and the porcine transmissible gastroenteritis virus (TGEV) in an actual sample, and can be used for the detection of the porcine intestinal coronavirus. The invention provides a novel and reliable technical method for trace early warning and monitoring of early-stage infection of four porcine intestinal coronavirus, differential diagnosis of other common viruses, research of propagation rules of the viruses, epidemiological risk assessment and other systematic applications.

The invention belongs to the technical field of gene detection of cross fusion of biology and a new generation of information technology, and particularly relates to a method for detecting residual DNA of multiple host cells based on digital PCR (Polymerase Chain Reaction). The invention discloses a multiplex digital PCR quantitative detection primer pair and a probe group for detecting residual DNA of CHO cells, Vero cells, pichia pastoris, escherichia coli and NS0 cells for the first time. By utilizing the method, not only can the residual DNA of five host cells be simultaneously subjected to single or multiple detection in a single reaction, but also non-specific amplification, signal interference and detection limitation of the residual DNA of the host cells in the qPCR detection process can be solved friendly, and meanwhile, the original concentration of a sample is absolutely quantified by the method; the method does not depend on the establishment of a standard curve, reduces the influence caused by a standard product, and is superior to the prior art.

The invention belongs to the technical field of analysis and testing, and discloses an MALDI (matrix-assisted laser desorption ionization) mass spectrum imaging probe for quantifying amino acid and a method for quantifying the amino acid. The probe is simple to synthesize and can quickly and efficiently react with an amino compound, and a product is stable and strong in ionization. The detection method comprises the steps of probe characterization, standard curve making and Q-MSI strategy, and absolute quantification of amino acid is realized by adopting an isotope labeling internal standard method and combining MALDI mass spectrum imaging. According to the method, parameters such as probe spraying, incubation and matrix spraying are optimized, so that the problem that spatial distribution information is lost in tissue homogenization in the prior art is solved, spatial distribution and content difference information of amino acid can be obtained, the sensitivity is improved by several times compared with that of self-detection of amino acid, and the method has good linearity and practicability and is suitable for popularization and application. The quantitative mass spectrum imaging strategy can also be popularized and used for quantitative analysis of other endogenous compounds.

The invention relates to a primer probe combination and a kit for detecting eDNA of Chinese sturgeons and application of the primer probe combination and the kit. Specific primers and TaqMan probes are designed for a D-loop region of Chinese sturgeon mitochondrial DNA, and efficient specific amplification and absolute quantification of Chinese sturgeon eDNA in an environmental sample are realized by optimizing a reaction system and amplification conditions. Compared with the prior art, the method is simple and convenient to operate, does not need to perform harmful sampling on Chinese sturgeon individuals, is suitable for resource monitoring and protection evaluation in a large-range and dynamic environment, and has high sensitivity, high accuracy and high practical value.

The invention relates to the field of antibodies, in particular to an anti-Der p 1 antibody and application thereof. The invention provides an anti-Der p 1 antibody, the anti-Der p 1 antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises an amino acid sequence of a complementarity determining region, and the amino acid sequence of the complementarity determining region comprises CDR1 as shown in SEQ ID No.1, CDR2 as shown in SEQ ID No.2 and CDR3 as shown in SEQ ID No.3; or the amino acid sequence of the heavy chain variable region is as shown in SEQ ID No.8. The anti-Der p1 antibody provided by the invention is a monoclonal antibody with high affinity and high sensitivity, after the monoclonal antibody is subjected to humanized IgE transformation, the monoclonal antibody is applied to a Dermatophagoides pteronyssinus allergen component Derp1 specificity IgE magnetic particle chemiluminescence quantitative detection reagent, and a standard curve independently calibrated by the Dermatophagoides pteronyssinus allergen component Derp1 can be obtained; real, accurate and repeatable absolute quantification is realized, the Dermatophagoides pteronyssinus allergen component Derp 1 specificity IgE with the concentration of 0.1 IU / mL can be detected, and the method has clinical diagnosis significance in allergic reaction identification.

The invention relates to a library construction method for UMI and Poly-A tail analysis and application of the library construction method. According to the invention, a novel library construction process is designed, a UMI + Poly-A tail tandem library process is established, all sequences of the Poly-A tail can be completely obtained, the method can be used for simultaneous sequencing analysis of UMI and Poly-A tail, absolute quantification can be carried out on original transcripts, the sequencing efficiency and precision are improved, short PCR products are connected in series, long fragments are formed, and the sequencing time is shortened. The advantages of long sequencing length and long reading length of Pacbio can be fully utilized, the most original RNA molecules are accurately backtracked and counted, and a real gene expression profile is obtained.

The present application relates to the technical field of molecular biology detection, in particular to an absolute antibody Taq DNA polymerase activity determination method based on real-time fluorescence quantitative detection, comprising the following steps: firstly, establishing a standard curve of DNA content and fluorescence value by reacting different concentrations of double-stranded lambda DNA with fluorescent dyes; then, performing an extension reaction in a specific reaction system using M13 single-stranded DNA as a template, recording fluorescence values in real time, calculating net fluorescence values and establishing a linear relationship between the net fluorescence values and reaction time; converting the amount of newly generated double-stranded DNA at different time points by using the standard curve, further calculating the consumption amount of dNTPs, and finally obtaining the absolute activity of the antibody Taq DNA polymerase according to the definition of the activity of the antibody Taq DNA polymerase. The whole process combines fluorescence detection and a standard curve to realize accurate determination of enzyme activity.

The invention discloses an RPA primer pair for absolute quantification of pseudomonas aeruginosa, double crRNA and a kit, and belongs to the fields of biology, detection and in-vitro diagnosis. The RPA primer pair provided by the invention is screened and designed for specific genes of pseudomonas aeruginosa, and can amplify target gene segments. The double crRNA is crRNA1 and crRNA2, and both the crRNA1 and the crRNA2 can specifically recognize corresponding target gene segments. The RPA primer pair, the double crRNA and the kit provided by the invention can realize absolute quantitative detection of pseudomonas aeruginosa, and have the characteristics of low cost (no need of expensive instruments, high sensitivity and the like); a portable constant-temperature fluorescence detector is adopted), the operation is simple and convenient (only 37 DEG C reaction is needed), the efficiency is high (the detection time is less than 1 hour), the sensitivity is high (the minimum is 10CFU), the resolution ratio is high (the quantification of the seed level is realized), a standard curve is linearly fitted based on inflection points, the accuracy is high (R2 is greater than 0.99), and the like.

Disclosed is a biological control composition comprising a known molar amount of bacilysin, or a known and high ratio of bacilysin to fengycin, uses of said composition, a method for purifying bacilysin and a method for absolute quantification of bacilysin.

The invention relates to the technical field of biology, and discloses a molecular beacon and a high-throughput sequencing library absolute quantification method thereof, and a fluorescent molecular beacon oligonucleotide (Oligo) sequence comprises a nucleotide chain (Loop chain) of a sequencing library recognition region, a first universal sequence region at the 5'upstream of the nucleotide chain (Loop chain) of the sequencing library recognition region, and a second universal sequence region at the 3 'downstream of the nucleotide chain (Loop chain) of the sequencing library recognition region; 5'of the first universal sequence region is modified into a fluorophore, and 3 'of the second universal sequence region is modified into a quenching group; and a nucleotide chain of the sequencing library recognition region is from an Illumina anmena sequencing platform or an MGI Huazai sequencing platform. The invention comprises a fluorescent molecular beacon for high-throughput sequencing library sequencing before-loading quantification, a sequence applied to a high-throughput sequencing platform and a detection method for library absolute quantification based on the molecular beacon, and overcomes the defects of a current gold standard detection method in technology and efficiency. The quantitative precision of the sequencing library is ensured; meanwhile, the economic and time cost of an actual application end is greatly reduced.

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 invention discloses a multiplex fluorescent quantitative PCR (Polymerase Chain Reaction) kit and application thereof. According to the invention, a multiplex fluorescent quantitative PCR (Polymerase Chain Reaction) detection method for detecting serotype 4h listeria monocytogenes, other listeria monocytogenes and listeria ivanovii is established through the design of primers of specific plcB and i-inlE and a double-labeled probe. According to the multiplex fluorescent quantitative PCR detection method, 4h L of listeria monocytogenes, other listeria monocytogenes and listeria ivanovii infection of serum can be distinguished, and the pathogenic bacteria in food can be quantitatively detected. According to the method, the constructed specific recombinant plasmid is used as a positive quantitative standard substance, absolute quantification of target nucleic acid is achieved through a standard curve, and the method has the advantages of being convenient to operate, high in specificity, high in sensitivity, good in repeatability and the like; the method is suitable for rapid qualitative and quantitative detection of listeria monocytogenes and listeria ivanovii in clinical, food and environmental samples.

This invention discloses a method for constructing a multi-target amplicon abundance standard for microorganisms and its application, belonging to the fields of molecular biology detection and microbiome analysis. This standard material screens sequences of common human gut microbiota strains, designs and adds universal primers, and obtains 14 DNA fragments of different lengths through gene synthesis, cloning, and purification, which are then mixed according to a preset abundance. It retains natural characteristics such as primer binding sites and GC content, and can systematically correct technical deviations in amplicon sequencing, solving the problem of species abundance distortion. This standard material functions as both a non-homologous internal reference and a homologous external reference, and can be used for laboratory proficiency testing, reagent kit performance evaluation, and cross-platform data calibration, promoting the leap from relative qualitative to absolute quantitative research in microbiome studies and providing metrological support for the standardization and precision of detection results.

The invention discloses a chiral compound quantification method based on an enantiomer internal standard, and belongs to the technical field of testing. According to the method, chiral distinguishing and enantiomer excess (ee) determination are carried out based on a diastereoisomerization strategy and ion mobility mass spectrometry, and absolute quantitative analysis of chiral compounds can be realized with extremely high flux by adding an enantiomer internal standard; the method can realize dual quantitative analysis of absolute concentration and stereocomposition of the chiral compound only by depending on mobility dimension, is successfully applied to high-throughput screening of yield and stereoselectivity of an enzyme catalytic reaction, can avoid interference of a matrix effect and an ion inhibition effect, and has the advantages of high analysis speed, high accuracy, simplicity, convenience and rapidness.

The application discloses a kind of absolute quantitative detection method of child respiratory tract multi-pathogen based on multi-target ladder microarray and exogenous internal reference correction.The method sets up independent micro-reaction unit to different pathogen, adds fixed amount of exogenous internal reference nucleic acid, and calculates template number by positive unit proportion combined with Poisson distribution model, then corrects the result by using the theoretical addition amount of exogenous internal reference, so as to realize parallel detection and absolute quantification of pathogen such as bocavirus, adenovirus, mycobacterium tuberculosis and mycoplasma pneumoniae without standard curve and special digital PCR equipment.The application has the advantages of high sensitivity, strong anti-interference ability, suitable for mixed infection sample analysis and clinical application, etc.

This invention discloses an absolute quantification method for pathogens and drug resistance genes in atmospheric bioaerosols and its application. Addressing the bottleneck of achieving absolute quantification of low-concentration samples while maintaining cell integrity and high-concentration nucleic acid extraction, this invention constructs a collection carrier by immobilizing a hydrophilic mixed cellulose ester microporous membrane on a technical agar plate. This is combined with a flow rate ≤30 L / min impactor sampler, ensuring structural integrity while capturing microorganisms, laying the foundation for flow cytometry quantification. This front-end collection system is deeply coupled with a dedicated cascade lysis and concentration process, enabling the acquisition of high-quality DNA suitable for metagenomic library construction and relative abundance analysis even under short-term, low-flow-rate collection. Simultaneously, the total microbial count is absolutely quantified using flow cytometry on the collected samples, and this result is fused with the relative abundance results. Through synergistic optimization of the entire chain, quantitative analysis of species-level pathogens and drug resistance genes in atmospheric bioaerosols is ultimately achieved.

The invention belongs to the technical field of medical biological detection, and particularly relates to a multiple microdroplet type digital PCR detection kit for central nervous system infection pathogens and application of the multiple microdroplet type digital PCR detection kit. The kit is based on the digital PCR technology, the detection range of the kit covers 23 kinds of clinically common pathogens of central nervous system infection of bacteria, fungi, viruses and mycoplasmas, and pathogen coverage is more complete. Compared with a real-time fluorescent PCR quantitative method, accurate absolute quantification can be realized without setting a standard curve. The kit has the advantages of wide detection range, accurate quantification, high sensitivity, simplicity and convenience in operation, controllable cost and the like, the detection rate of the central nervous system infection pathogens is greatly increased, and an accurate and rapid judgment basis is provided for early diagnosis and early treatment of central nervous system infection.

The invention develops a method for absolute quantification of fungal population based on a Pacbio three-generation long fragment sequencing platform and application thereof, and belongs to the technical field of microbial amplicon detection. According to the method, the synthesized chimeric DNA is used as an internal standard, and a fungal community in a sample is absolutely quantified on the basis of an amplicon high-throughput sequencing platform. The specific method comprises the following steps: directly adding the synthesized chimeric DNA into an environmental sample; extracting genome DNA of the environment sample; carrying out high-throughput sequencing on the ITS full-length sequence of the fungus through PCR amplification; the high-throughput sequencing results calculate the relative abundance and absolute abundance of the amplicon family (e.g., the copy number of fungal ITS per unit mass sample). A highly complex environmental sample is taken as a research object to prove that the method can sensitively and accurately quantify the absolute abundance of a specific group. The method is widely applicable to microbial community analysis of samples from different sources, including environmental samples from soil, samples from plant tissues, samples from human intestinal tracts, and samples from food samples. The method disclosed by the invention is simple to operate, accurate in detection and wide in application, and has a wide market prospect in identification and analysis of microbial communities.

The invention discloses a method for absolute quantitative construction of synthetic flora and application, and relates to the technical field of microbial control. According to the invention, a microbial combination is screened by virtue of an absolute quantitative method of qPCR (quantitative polymerase chain reaction), an optimal synergistic ratio is determined by virtue of a multi-ratio inoculation experiment to construct a synthetic flora, the synthetic flora comprises biocontrol bacteria Trichoderma spiraloides RS05, Bacillus siamensis B11 and Streptomyces mirabilis BD2233, the biocontrol bacteria are mixed according to a ratio of 3: 2: 1, and the biocontrol bacteria are mixed according to a ratio of 3: 2: 1. The bacterial strain can significantly improve the colonization efficiency and growth promotion effect of microorganisms in plant rhizosphere, can be used as a synthetic bacterial agent for promoting plant growth, especially has a significant promotion effect on growth of Zhuang * green hybrid bamboo, and has an application prospect in the field of planting of Zhuang * green hybrid bamboo.

This invention discloses a single-cell microarray-based kit and method for in situ quantitative detection of receptor-binding allergen-specific IgE. The kit includes: a single-cell microcavity array chip, an antigen barcode chip, a plastic clamp, IgE standards, fluorescently modified IgE detection antibodies, and auxiliary reagents. During detection, basophil single cells are loaded into the microcavity array chip, lysis buffer is added, and the cells are clamped together with the antigen barcode chip. Low-temperature incubation allows the allergen-specific IgE released from cell lysis to be captured in situ by the antigen on the chip. A sandwich immunoassay is then performed using a fluorescently labeled antibody, and absolute quantification is achieved using a standard curve. This invention is the first to achieve high-throughput, high-sensitivity, multi-component parallel in situ quantitative detection of basophil membrane receptor-binding sIgE at the single-cell level. It can directly reflect the functional sensitization state of effector cells and has advantages such as simple operation, low sample requirement, and no need for complex valve devices.

The invention relates to the field of allergen detection, in particular to an anti-Der p 2 antibody and application thereof. The invention provides an anti-Der p 2 antibody. A heavy chain variable region of the anti-Der p 2 antibody comprises an amino acid sequence of a complementarity determining region: CDR1 as shown in SEQ ID No.1, CDR2 as shown in SEQ ID No.2, and CDR3 as shown in SEQ ID No.3; or the amino acid sequence of the heavy chain variable region is as shown in SEQ ID No.8. According to the protein, a New Zealand rabbit is immunized by utilizing Der p 2 protein of a main allergen of Dermatophagoides pteronyssinus, and a group of monoclonal antibodies with high affinity and high sensitivity to Der p 2 are obtained through screening. The antibody is further subjected to humanized IgE modification and is applied to a magnetic particle chemiluminescence quantitative detection reagent of Der p 2 specific IgE. According to the reagent, a standard curve for Der p 2 independent calibration can be established, so that accurate and repeatable absolute quantitative analysis is realized, and the detection sensitivity can reach 0.1 IU / mL. The detection method formed on the basis of the anti-Der p 2 antibody provided by the invention has important value for clinical diagnosis of anaphylactic reaction.

The invention discloses a metagenome activity quantification method based on an exogenous artificially synthesized endogenous reference gene and application of the metagenome activity quantification method, and relates to the technical field of biology. According to the invention, an endogenous reference gene as shown in SEQ ID NO.1 is firstly designed, and the endogenous reference gene has no homology with an existing organism and can realize homology interference. On the basis, a metagenome activity quantitative method is further designed, and the core thought of the metagenome activity quantitative method comprises the following steps: providing an artificially synthesized vector containing an endogenous reference gene, setting the artificially synthesized vector as a standard system with gradient concentration, and adding the standard system into a sample to be detected; before addition, a to-be-detected sample is pretreated through light-sensitive DNA combined with dye so as to shield dead bacteria DNA interference, then nucleic acid extraction, library construction and sequencing are carried out, and metagenome activity quantification is obtained through bioinformatics analysis and calculation. According to the method, homology interference is eliminated from the source, absolute quantification of microbial activity is realized, the stability and standardization degree of an internal standard are improved, and the method is suitable for quantitative analysis of metagenomes in various scenes.

The invention discloses an absolute quantitative detection method for urine exfoliated podocyte, relates to the technical field of biomedical detection, and aims to solve the problem that the urine exfoliated podocyte cannot be accurately and absolutely quantified due to uncertain cell loss in a sample treatment process in the prior art. The method comprises the following steps: before carrying out any physical treatment on a urine sample, adding a known number of internal standard reference substances with physical characteristics similar to those of cells into the urine sample, carrying out co-enrichment treatment on a mixed sample containing target podocytes and the internal standard reference substances, and preparing a cell slide by adopting a standardized slide preparation technology; and carrying out podocyte specific immunofluorescence staining on the slide. Through the design of internal standard preposition and whole-course synchronous calibration, the cell loss error in the operation process is effectively overcome, the traditional semi-quantitative detection is improved into accurate and repeatable absolute quantification, and the reliability and clinical application value of a detection result are remarkably improved.

The application belongs to the field of gene sequencing, and discloses a microdroplet digital PCR absolute calibration method for mixed standard substances for high-throughput sequencer calibration and application thereof. The method comprises the following steps: providing N sets of primer probes for N targets in the standard substance; full background specificity verification: for the i th set of primer probes, it is necessary to prove that it can specifically detect the i th target, and prove that it has no specific detection signal in the mixed DNA sample composed of the remaining N-1 targets; only using all the primer probes verified in S2, performing parallel digital PCR detection on the standard substance sample; and calculating the absolute copy number concentration of each target according to the detection result. The method ensures the quantitative "purity" of each target in a complex mixed background, provides guarantee for high-confidence calibration results from the process design, and improves the throughput and efficiency of multi-target detection.

The application discloses a nucleic acid absolute quantification method based on nucleic acid constant temperature amplification, which comprises the following specific steps: step one, sample pretreatment, blood samples and saliva samples are pretreated respectively; step two, reaction system configuration and assembly; step three, digital micro-reaction unit preparation and amplification; and step four, multiple signal analysis and absolute quantification. In the application, multiple primer design and a double-probe system are adopted, so that the mutation detection rate is improved, and the detection limit can be reduced to 0.1% variation through Cas12a-sgRNA auxiliary verification. The double-emulsification technology of the micro-fluidic chip is adopted, so that the droplet diameter variation coefficient is controlled within 5%, and the clogged micropore rate is reduced by real-time analysis of the micropore array image through the ImageJ software, automatic marking and exclusion of abnormal units.

The application belongs to the technical field of molecular diagnosis, and particularly relates to a digital PCR kit and method for accurately detecting the copy number of a CAR gene of a CAR-T cell and a lentivirus vector. The application first realizes the absolute quantification of CAR, Rev and an internal reference gene RNase P in a single tube simultaneously, and has strong universality. Based on the principle of digital PCR, the absolute copy number is directly obtained without a standard curve, and the result is accurate and highly repeatable. The detection lower limit reaches 0.01%, and extremely low abundance CAR-T residues or microresidual diseases can be detected. Primers and probes are designed for the conservative regions of CD3zeta, Rev and RNase P, and no cross reaction is verified, so that false positives are effectively avoided. The sample only needs gDNA, and there is no requirement for cell activity, and the sample is compatible with freezing and transportation. The kit is premixed and optimized, the process is automated, and 4-5 hours are needed. The kit is suitable for the whole chain of CAR-T drug research and development, production process quality control, clinical patient monitoring and safety evaluation.

The present application relates to a kind of based on peptide segment sensitization's high-precision scale absolute quantification method of proteome.It is using chemical derivatization technique to be modified on peptide segment, to change the physicochemical property of peptide segment, improve its mass spectrum signal response intensity.The derivatization sensitizer used generally includes positive charge or proton affinity group, it is helpful to improve the ionization efficiency of peptide segment.Due to the difference of the physicochemical property of peptide segment itself, after being labeled by the same derivatization sensitizer, usually low-abundance peptide segment signal response greatly improves, and high response peptide segment changes less, to cause the difference of the mass spectrum response signal between peptide segment to reduce, so that intensity information can more accurately reflect the content of peptide segment.The advantage of this method is: high accuracy of quantification, can realize large-scale proteome absolute quantification analysis.

The invention relates to the technical field of gene engineering, and particularly discloses a construction method and application of intracellular stable self-expression pH-sensitive fluorescent protein particles. The recombinant plasmid comprises a coding gene of a pH sensitive fluorescent protein pHlurin and a J23119 promoter; the preparation method comprises the following steps: cloning a pH sensitive fluorescent protein pHlurin gene segment to a pBAD24 plasmid, and replacing an araBAD promoter of the pBAD24 plasmid with J23119; the nucleotide sequence of the pHlurin gene is as shown in SEQ ID No. 1; the nucleotide sequence of the J23119 is as shown in SEQ ID No. 2. The recombinant plasmid constructed by the invention not only realizes absolute quantification of intracellular pH, but also has better detection performance and accuracy; in addition, the application environment of the plasmid is expanded, and continuous, stable and high-expression of the pH-sensitive fluorescent protein can be realized without an inducer.

The present invention provides a primer-probe combination for detecting HPV and / or HPV typing, belonging to the field of human papillomavirus (HPV) detection technology. The primer-probe combination includes one or more of HPV16E7, HPV18E7, HPV31E7, HPV45E7, HPV33E7, HPV35E7, HPV59E6, HPV68E6, HPV58E7, HPV51E6, HPV52E7, HPV56E6, HPV66E6, and HPV39E6, and can distinguish different types of HPV, with good specificity, accuracy, and low detection limit. The present invention uses a digital PCR method (dPCR) to detect the fluorescent signal of each droplet, and calculates the copy number of the target gene fragment in the entire reaction system based on the Poisson distribution principle, so that absolute quantification can be achieved without relying on a standard curve.

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.