199 results about "Absolute quantification" patented technology

The invention discloses a gradient internal reference-based absolute quantification method for plant endophyte bacteria, which comprises the following steps of: designing a specific primer for amplifying a plant endophyte bacteria absolute quantification internal reference sequence of a part matched with conserved sequences on two wings of an internal reference sequence; the method comprises the following steps: obtaining an internal reference sequence for amplifying absolute quantification of plant endophyte bacteria and increasing conserved sequences on two wings, and a specific primer for amplifying the plant endophyte bacteria 16SV5V7, mixing the series of internal reference sequences according to a certain gradient proportion, and jointly amplifying and building a library together with total DNA of a plant tissue sample, so as to obtain a specific primer for amplifying the plant endophyte bacteria 16SV5V7. The abundance of the endophyte flora in the plant tissue sample is absolutely quantified, so that the condition of quantitative distortion in plant endophyte detection is eliminated.

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 biological detection, and relates to a method for detecting lentivirus titer by a double-standard-curve RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) method. Infecting the target cells by adopting a to-be-detected sample containing a recombinant lentivirus, and collecting and extracting genome DNA of the infected target cells; carrying out a viral genome RT-qPCR amplification reaction and an internal reference genome RT-qPCR amplification reaction by respectively taking the original virus standard substance and the internal reference standard substance which are diluted in a gradient manner and the extracted target cell genome DNA as templates; and calculating the lentivirus titer corresponding to the sample according to data obtained by the RT-qPCR amplification reaction. The method can be used for absolute quantitative qPCR detection of lentivirus titer, can accurately determine lentivirus successfully integrated into a cell genome, and reflects the real infection ability of the virus; in addition, the technology has high sensitivity and specificity, is not influenced by the extraction efficiency of the cell genome DNA, and ensures the reliability of the detection result.

A method for the absolute quantification of an exosome surface protein and the use thereof. The method for the absolute quantification of the exosome surface protein comprises: (1) extracting an exosome containing a target surface protein using a solid-phase support material coupled with an antibody which identifies the target surface protein, sequentially performing exosome lysis and desalting, and preparing the desalted protein into a sample to be detected; (2) analyzing the sample to be detected using HPLC-PRM / MS; selecting a characteristic peptide fragment in the target surface protein as an internal standard, labeling the characteristic peptide fragment with a heavy isotope to obtain a heavy peptide, using the characteristic peptide fragment without isotope labeling as a light peptide, diluting the synthesized light peptide into different concentration gradients, diluting the synthesized heavy peptide into a fixed concentration, and mixing the two at a volume ratio of 1:1 to obtain a standard; and (3) establishing a standard curve of the internal standard, and performing the absolute quantification of the exosome surface protein according to the standard curve. The method for the absolute quantification of the exosome surface protein has high sensitivity, good reproducibility and high accuracy, and the quantitative range can reach 5 orders of magnitude.

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 the technical field of gene detection, in particular to a primer and probe combination and a kit for detecting human papilloma virus nucleic acid 14 typing based on a digital PCR (polymerase chain reaction) method and application of the primer and probe combination and the kit. 14 HPV high-risk types including human papilloma viruses HPV52, HPV35, HPV18, HPV66, HPV39, HPV56, HPV68, HPV16, HPV31, HPV58, HPV59, HPV33, HPV45 and HPV51 can be quantitatively detected by utilizing a micro-droplet digital PCR method, compared with the prior art, the quantitative detection of more HPV types can be realized, the specificity is strong, the sensitivity is high, the lowest detection limit is reduced to 0.1 copies / uL, and real absolute quantification is realized.

The invention relates to the technical field of sample detection, in particular to a qualitative and quantitative detection method of M protein, electronic equipment and a storage medium. The qualitative and quantitative detection method for the M protein comprises the following steps: acquiring a mass spectrum of a sample to be detected; the mass spectrum at least comprises a total light chain region; carrying out peak value detection on the mass spectrum, and judging whether M protein peaks exist in the total light chain region or not; and for the mass spectrum with the M protein peak, calculating the curve lower peak area SM of the M protein peak and the curve lower peak area SLC of the total light chain region, and combining a serum total immunoglobulin quantitative result or a serum total light chain quantitative result to obtain the absolute content MW of the M protein. The method provided by the invention can realize absolute quantitative detection of the M protein, and solves the problem that absolute quantitative detection cannot be carried out on the M protein in the prior art.

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.

The capabilities of gas or liquid chromatography-molecular rotational resonance (GC / LC-MRR) instrumentation exceed those of high-resolution mass spectrometry and nuclear magnetic resonance in terms of selectivity, resolution, and compound identification. MRR detection provides high specificity for selective gas or liquid phase separations, including the ability to resolve co-eluting peaks and isomeric compounds without any loss of specificity or accuracy. MRR allows for qualitative identification and absolute quantification of analyte components separated by GC or LC in the absence of reference standards. GC-MRR is ideal for compound-specific isotope analysis (CSIA) and can identify enantiomers and enantiomeric excess. GC-MRR measurements are particularly useful for studying biosynthesis / degradation and geochemical isotopic compounds.

The present invention relates to an absolute quantitative analysis method and application of a key metabolite of methionine cycle. The present invention screens an efficient reduction-sulfhydryl derivatization scheme, and establishes an HPLC-MS / MS detection platform. Deuterated phenylalanine is used as an internal standard to draw detection standard curves of 9 key metabolites of methionine cycle, including methionine, homocysteine, cysteine, choline, betaine, reduced glutathione, S-adenosylhomocysteine, S-adenosylmethionine, and glutamic acid, and the linearity and range, precision and accuracy, stability, recovery rate, and matrix effect of the detection platform are investigated. The investigation results show that the quantitative analysis method established by the present invention has high detection stability and accuracy for key metabolites of methionine cycle, especially for homocysteine, cysteine, and reduced glutathione, which have low detection sensitivity and are easily damaged by oxidation, and the detection sensitivity is increased by 14.44 times, 5.08 times, and 3.41 times, respectively, and the stability is greatly improved.

The present invention discloses a crRNA for detecting Epstein-Barr virus based on microdroplet digital analysis, wherein the crRNA is selected from one or more of the crRNAs shown in nucleotide sequences such as SEQ ID NO: 1 to SEQ ID NO: 11. The quantitative detection kit for Epstein-Barr virus nucleic acid constructed using the crRNA of the present invention does not require a DNA amplification step in the process of detecting Epstein-Barr virus, thus avoiding the result bias and possible contamination risk generated during the amplification process; and the result collection is completed using a fluorescent microscope, which is convenient and fast, and is suitable for rapid detection at the screening site. In addition, the quantitative detection kit for Epstein-Barr virus nucleic acid can achieve absolute quantification of the single-copy Epstein-Barr virus DNA level, without the need for internal reference correction and the establishment of a standard curve, thus avoiding the error caused by the use of different standards, and compared with the traditional qPCR detection method, the screening results have significant advantages.

The invention discloses a construction method of a multiple RT-qPCRTaqMan probe absolute quantitative detection system. The construction method comprises the following steps: (1) constructing a pACYC-MS2 recombinant plasmid; (2), pACYC-MS2-lncRNAs (deoxyribonucleic acids) are selected from the group consisting (3) verifying the accuracy of the pACYC-MS2-lncRNAs recombinant plasmid by sequencing: converting the pACYC-MS2-lncRNAs into BL21 (DE3) escherichia coli, performing induced expression, performing PEG enrichment, performing molecular sieve chromatography purification on MS2VLPs, and performing verification and performance evaluation; (4) constructing a one-step method multiple RT-qPCRTaqMan probe detection system; and (5) establishing a multiple RT-qPCRTaqMan probe absolute quantitative detection system. On the basis of the purpose of absolute quantification of circulating lncRNA in plasma, a novel detection method is established by adopting one-step RT-qPCR, reverse transcription and PCR are completed in the same reaction tube, operations such as uncovering, pipetting and sample adding do not exist in the midway, the experimental efficiency is greatly improved, and the pollution risk of the reaction is reduced.

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.

Provided herein are methods and systems and related composition, to provide a rodent model having a target microbiome profile formed by a target presence, a target proportion and / or a target total load of a target prokaryote of a target taxon, based on absolute quantification of the target prokaryote. Further provided are rodents obtained by the methods herein described and related use in testing methods performed in connection with physiological or pathological conditions in an individual preferably a human individual. Also provided herein is a tailcup device and related use to prevent coprophagia in rodents.

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.