64 results about "Indirect immunofluorescence" patented technology

The invention provides a full-automatic indirect immunofluorescence interpretation method and system, a storage medium and a program product, and relates to the technical field of indirect immunofluorescence detection.The method comprises the steps that when identification information of a target sample is obtained, a plurality of sealed antigen slides are placed in slide carrying groove positions of a bearing platform in batches, and the target sample is obtained; driving the bearing platform through the transmission mechanism to sequentially move the plurality of antigen slides to the imaging area, and performing positioning and fixing operation on the antigen slides entering the imaging area by using the positioning and clamping device; sequentially carrying out multi-view scanning on the antigen slides entering the imaging area to obtain multi-view fluorescence images, and splicing the fluorescence images to generate a fluorescence panoramic image; performing feature extraction on the panoramic image, determining a fluorescence karyotype category and a target fluorescence intensity value, and determining an antibody titer according to the category, the intensity value and dilution information; and performing association mapping on the titer, the category and the panoramic image according to the identification information to generate a fluorescence interpretation result.

The invention discloses an application of an AP2-F2 protein polyclonal antibody as a cryptosporidium female gamete positioning tag antibody. The AP2-F2 protein of cryptosporidium parvum is subjected to truncated prokaryotic expression, protein is purified to immunize an ICR mouse, a polyclonal antibody is prepared, ELISA and WB detection of antibody titer are carried out to prove that the polyclonal antibody is effective, then indirect immunofluorescence detection of the antibody is carried out to determine that the polyclonal antibody is located on the female gametes of cryptosporidium parvum, and the polyclonal antibody can be used as a label to locate the female gametes of cryptosporidium parvum. The method plays an important role in localization research of unknown protein of cryptosporidium.

The invention relates to the technical field of general image data processing or generation, in particular to a multi-view indirect immunofluorescence medical image classification interpretation method and system, and the method comprises the steps: mapping an ethanol view, a formaldehyde view and an HEp-2 view to a feature space, and converting into a feature sequence; performing position coding on the feature sequence to obtain position information; based on the feature sequence with the position information, modal identification information is added, and an identification feature sequence is determined; based on the identification feature sequence, executing multi-modal information fusion, and determining a multi-modal feature sequence; based on the multi-modal feature sequence, combining an attention mechanism to determine a long-distance dependency relationship and correlation between the data; and based on the attention mechanism model, outputting the probability of each category to execute classification interpretation. According to the method, the type of the anti-neutrophil cytoplasm antibody fluorescence model can be adaptively and comprehensively judged in combination with the characteristics of fluorescence brightness, forms and the like of different views, and interference existing in a single view is avoided.

The invention belongs to the technical field of biomedicine, and discloses an antinuclear antibody IgG indirect immunofluorescence detection method and system, and the detection standardization degree and the result reproducibility are significantly improved through the full-automatic system design in combination with precise mechanical transmission and an automatic sample adding device. Through an optimized FITC labeling system and a refined washing procedure, non-specific binding is remarkably reduced, and meanwhile, the binding efficiency of the antinuclear antibody and the secondary antibody is improved, so that the detection sensitivity is improved. Through the design of the automatic sample adding and sheet sealing device, the sample treatment efficiency is greatly improved, and simultaneous detection of large-scale samples can be realized. Through combination of a high-resolution imaging system and an intelligent image analysis algorithm, morphological characteristics and distribution rules of fluorescence signals are automatically extracted, and the accuracy and objectivity of a detection result are greatly improved.

The application discloses a hybridoma cell strain 6F92E10, a monoclonal antibody thereof and application thereof, and belongs to the technical field of biotechnology.The application provides a hybridoma cell strain secreting an anti-mucinophilic Akkermansia monoclonal antibody.The antibody secreted by the hybridoma cell strain only combines with AKKermansia and does not cross-react with other common intestinal bacteria.In addition, the application solves how to detect the adhesion and colonization of mucinophilic Akkermansia in the colon of a mouse by using the prepared monoclonal antibody through an indirect immunofluorescence localization experiment, and expands the application range of the antibody.

The application belongs to the technical field of biology and particularly relates to an African swine fever virus pNP868R protein monoclonal antibody and a preparation method and application thereof. The application takes the pNP868R protein prepared by eukaryotic expression as an antigen, immunizes BALB / c mice, uses hybridoma technology, and obtains a hybridoma cell line capable of stably secreting the anti-African swine fever virus pNP868R protein monoclonal antibody through cell fusion, indirect ELISA method screening, limited dilution method subcloning and hybridoma continuous passage culture. The application also obtains an African swine fever virus pNP868R protein monoclonal antibody. The monoclonal antibody has high affinity with the pNP868R protein and can be applied in Western-Blot experiments, indirect immunofluorescence experiments, immunoprecipitation experiments and the capture of proteins interacting with pNP868R, and can be used for the diagnosis, prevention or treatment of African swine fever.

The invention provides an anti-double-stranded DNA antibody indirect immunofluorescence method picture interpretation method and device and a medium, and the method comprises the following steps: obtaining a to-be-collected slide which is obtained by treating a to-be-detected serum sample through an anti-double-stranded DNA indirect immunofluorescence method; performing image acquisition on the to-be-acquired slide to obtain an acquired image set corresponding to the to-be-acquired slide; performing image processing on the acquired image set to obtain a first target image set; performing image processing on the first target image set to obtain a cell region corresponding to each first target image in the first target image set; and performing identification processing on the cell region corresponding to each first target image to obtain a sample report of the to-be-collected slide. According to the method, a series of steps of image acquisition, processing, cell region identification and the like are adopted for processing, so that the traditional manual operation is replaced, the automation of the interpretation process is realized, and the interpretation accuracy and efficiency are improved.

The invention belongs to the technical field of plant pharmacy, and particularly relates to an effect of pigeonpea in resisting respiratory syncytial virus infection. Detecting that the pigeon pea protein extract is non-toxic to HEp-2 cells through a cell proliferation detection kit; a cell activity detection method and an indirect immunofluorescence method are adopted to determine that the pigeon pea protein extract has an obvious inhibition effect on the respiratory syncytial virus, the activity of inhibiting the respiratory syncytial virus after the pigeon pea protein is subjected to enzymolysis by alkaline protease is improved, and pigeon peas have relatively high biological safety and relatively strong respiratory syncytial virus infection resistance.

The invention discloses application of ailanone in preparation of a medicine for treating feline infectious peritonitis, and belongs to the field of biological medicine. The change of the expression quantity of the feline infectious peritonitis virus N protein is detected through indirect immunofluorescence and western blot, and the change of the quantity titer of the feline infectious peritonitis virus is detected through TCID50. Results show that ailanone has an obvious effect of resisting the feline infectious peritonitis virus, the inhibitory effect on the feline infectious peritonitis virus is enhanced along with increase of the drug concentration, and the half effective concentration is 0.09 mu M. Meanwhile, drug inhibition tests show that ailanone has no inhibition effect on feline calicivirus. The invention provides a novel therapeutic drug and strategy for treating feline infectious peritonitis.

The invention belongs to the technical field of bioengineering, and particularly relates to a preparation method and application of an N protein polyclonal antibody of a horse coronavirus. The polyclonal antibody is prepared by truncation expression of the horse coronavirus N protein, and a material is provided for research on inhibition of host innate immune mechanism by the horse coronavirus N protein. Prokaryotic expression is carried out on the N protein of the equine coronavirus, the polyclonal antibody is prepared, and a method and a material are provided for molecular research and mechanism research of the equine coronavirus. According to the invention, in combination with bioinformatics analysis of an N protein of the horse coronavirus, a pGEX-6p-N3 recombinant plasmid is constructed, and prokaryotic expression is carried out, so that a purified recombinant protein is obtained. After a mouse is immunized by the recombinant protein, a polyclonal antibody is obtained, an indirect ELISA method detects that the polyclonal antibody has good titer, Western Blot detection has good reactivity, and the polyclonal antibody can be applied to an indirect immunofluorescence test.

The invention provides an application and a method of an RSAD2 agonist glycopyrronium in resisting RSV infection. The method comprises the following steps: S1, virtual screening and molecular docking; s1.1, performing virtual screening and molecular docking; s1.2, carrying out molecular dynamics simulation; s2, cell culture and RSV infection; s3, performing RT-qPCR detection on the expression of the RSAD2 and the virus load of the RSV; s4, performing Westernblot analysis on the influence of a small molecule compound on the expression of the RSAD2; s5, CCK8 is used for detecting the toxicity of the compound to Beaas-2b cells; s6, carrying out indirect immunofluorescence detection on a fluorescence signal of the RSV; s7, animal model establishment; s8, carrying out HE dyeing; s9, carrying out enzyme-linked immunosorbent assay (ELISA); and S10, statistical analysis. According to the present invention, the small molecule agonist targeting the host immune factor RSAD2 is screened, the endogenous antiviral factor is activated from the host perspective, the drug resistance problem caused by direct virus targeting is avoided, the RSAD2 is adopted as the host natural antiviral molecule, the activation strategy has good targeting property and good safety, and the efficient anti-RSV effect under the low toxic-side effect is achieved.

The present invention discloses a monoclonal antibody 2D1B1 against canine distemper virus H protein and its application, belonging to the technical field of monoclonal antibodies. The present invention screened out 9 monoclonal antibodies by indirect ELISA and indirect immunofluorescence identification. After indirect immunofluorescence identification, the 9 monoclonal antibodies obtained were all able to react with the vaccine strain (CDV-Onderstepoort). The results of Western Blot tests showed that the 9 monoclonal antibodies screened out were all able to react specifically with the eukaryotic expressed CDV H protein, among which 2D1B1 was also able to react specifically with the prokaryotic expressed CDV H protein, that is, 2D1B1 was not only able to react with the attenuated vaccine strain and the virulent isolated strain, but also with both eukaryotic and prokaryotic expressed CDV H proteins, and at the same time had a high neutralizing potency (2 6 ), which can be used to prepare broad-spectrum detection reagents and therapeutic preparations for identifying canine distemper virus H protein.

The invention discloses an in-vitro transcription system of theileria annulata transformed cells and a construction method of the in-vitro transcription system, the in-vitro transcription system comprises IVT-EGFP-mRNA, and the structure of the IVT-EGFP-mRNA is 5 'UTR-Cl-Kozak-EGFP-Pacl-3' UTR-polyA (120). According to the invention, mRNA for coding the EGFP gene is synthesized through in-vitro transcription, through capping, purification and agarose gel electrophoresis analysis, the expression condition of the target gene in TaNM1 cells is finally verified through an in-vitro transfection test and an indirect immunofluorescence technology, and the result shows that the in-vitro transcription system transfected and coded with the EGFP successfully expresses the EGFP, and the expression condition of the target gene in the TaNM1 cells is verified. The invention shows that a system for transfecting the EGFP gene into the theileria annulata transformed cell TaNM1 through an in-vitro transcription method is successfully established, and an important reference value is provided for design and research and development of theileria annulata RNA vaccines.

The application is based on machine learning model, and the polypeptide WFKDWY (166953) with potential is screened through polypeptide virtual screening technology based on the analysis of PEDV Nsp5 protein crystal structure. The sequence of 166953 is artificially synthesized by solid phase, and the polypeptide is screened by using artificially expressed PEDV Nsp5 protein through ELISA experiment. The affinity constant of the polypeptide and the target protein is identified by surface plasmon resonance experiment (SPR). The cytotoxicity of the polypeptide 166953 is tested by CCK-8 kit. The virus infection inhibiting activity of the polypeptide is tested by fluorescence quantitative PCR and indirect immunofluorescence experiment. The results show that the sequence 166953 can significantly inhibit the infection of PEDV.

This invention utilizes a machine learning model to analyze the crystal structure of the PEDV Nsp5 protein. Through virtual peptide screening technology, a peptide sequence with potential affinity for the target protein, YKKLHK(162860), was artificially synthesized in a solid phase. Using artificially expressed PEDV Nsp5 protein, the peptide was screened using an ELISA assay. The affinity constant between the peptide and the target protein was determined using surface plasmon resonance (SPR) assays. The cytotoxicity of peptide 162860 was tested using a CCK-8 assay kit. Its inhibitory activity against viral infection was tested using quantitative real-time PCR and indirect immunofluorescence assays. The results showed that the 162860 sequence significantly inhibited PEDV infection.

The invention relates to the technical field of glass slides for biomedical detection, and particularly discloses a high-adhesion type glass slide for indirect immunofluorescence analysis experiments. The glass slide comprises a substrate, a transition layer and a functional layer, the transition layer is arranged on the surface of the substrate, is made of metal oxide and is used for improving the bonding stability of the substrate and the functional layer; the functional layer is arranged on the surface of the transition layer and is prepared by compounding modified polyacrylamide containing amino / carboxyl and nano-particles, and the sample adhesion is synergistically enhanced through specific combination of functional groups and a nanoscale surface rough structure. Performance tests show that the glass slide has an adhesion rate of more than or equal to 95% on cell and tissue slices, washing resistance of more than or equal to 5 times and a fluorescence signal variable coefficient of less than or equal to 5%, can effectively solve the problems that the existing glass slide sample is easy to fall off and poor in detection repeatability, remarkably improves the accuracy of indirect immunofluorescence analysis, and is suitable for scenes such as clinical diagnosis and pathological detection.

The present invention discloses a canine distemper virus H protein monoclonal antibody 6A4A6 and its application, which belongs to the field of monoclonal antibody technology. The present invention screened out 9 monoclonal antibodies through indirect ELISA and indirect immunofluorescence identification, and named them 2C1A1, 2D1B1, 6A4A5, 6A4A6, 6G4H1, 6G4H3, 6H4F4, 6H4H5 and 6H4H6 respectively. After indirect immunofluorescence identification, the 9 monoclonal antibodies obtained were all able to react with the vaccine strain (CDV-Onderstepoort); the neutralizing antibody test results showed that among the 9 monoclonal antibodies obtained, 6A4A5 and 6A4A6 did not react with giant panda / SX / 2014 (a highly toxic isolate), but only reacted with the vaccine strain, while 6A4A6 had a higher neutralizing titer than 6A4A5 (up to 2 6 ), which can be used to prepare detection reagents and therapeutic preparations for identifying canine distemper virus vaccine strains.

The application is used for analyzing the S1-CTD region of the PEDV S protein crystal structure by means of a machine learning model, and a polypeptide sequence FWKSKR (110766) with potential is screened from the target protein by polypeptide virtual screening technology. The 110766 sequence is artificially synthesized on a solid phase, and the polypeptide is screened by means of ELISA experiment using artificially expressed PEDV S1 protein. The affinity constant of the polypeptide and the target protein is identified by means of surface plasmon resonance experiment (SPR). The cytotoxicity of the polypeptide 110766 is tested by CCK-8 kit. The virus infection inhibiting activity of the polypeptide is tested by means of fluorescent quantitative PCR and indirect immunofluorescence experiment. The results show that the 110766 sequence can significantly inhibit the infection of PEDV.

The application discloses a kind of H9N2 subtype AIV multi-copy M2e protein monoclonal antibody, heavy chain and light chain variable region respectively have the amino acid sequence of sequence table SEQ.ID.NO.1 and SEQ.ID.NO.2.The related cell line and its preparation method are also established according to this.The preservation number of hybridoma cell strain 4E10-A10 of the application is CCTCC NO:C202328, and test shows that the anti-multi-copy M2e protein monoclonal antibody secreted by it has high specificity, sensitivity, can specifically recognize 2M2e, 3M2e and 4M2e recombinant protein and the cell expressing H9N2 subtype avian influenza virus 3M2e protein, and is suitable for indirect immunofluorescence detection and Western-blot identification.In conclusion, the application provides material and technical support for the mechanism research of AIV M2e monoclonal antibody to AIV and the application effect evaluation of broad-spectrum vaccine taking AIV M2e protein as target.

This invention utilizes a machine learning model to analyze the S1-CTD region of the PEDV S protein crystal structure. Through virtual peptide screening, a peptide sequence of FWKEAK (110616) with potential affinity for the target protein was identified. The 110616 sequence was artificially synthesized in a solid phase, and the peptide was screened using ELISA with artificially expressed PEDV S1 protein. The affinity constant between the peptide and the target protein was determined using surface plasmon resonance (SPR) assays. The cytotoxicity of peptide 110616 was tested using a CCK-8 assay kit. Its inhibitory activity against viral infection was assessed using quantitative real-time PCR and indirect immunofluorescence assays. The results showed that the 110616 sequence significantly inhibited PEDV infection.

The invention discloses a monoclonal antibody and application thereof. The invention relates to a monoclonal antibody which is secreted by a hybridoma cell strain 6B12G10, and an identified antigen epitope is aa130-aa134 of gene I type African swine fever virus P30 protein. And aa131 amino acid of the P30 protein is E. According to the application, a monoclonal antibody capable of identifying a gene I type African swine fever virus P30 antigen is prepared; the monoclonal antibody can directly distinguish a gene type I, a gene type II and a gene I / II recombinant ASFV by utilizing an indirect immunofluorescence technology; for typing identification after ASFV low virulent strain infection, a blocking ELISA method is established by using the monoclonal antibody, whether gene type I ASFV infection or gene type II and gene I / II recombinant ASFV infection exists is effectively distinguished for the first time, the blank of differential diagnosis in the aspect at home and abroad is effectively filled up, and the monoclonal antibody has important significance on gene type I ASFV infection traceability.

The invention discloses a novel anti-echinococcus monoclonal antibody, a hybridoma cell strain and application of the novel anti-echinococcus monoclonal antibody, and belongs to the technical field of biomedicine and immunology. The preservation number of the hybridoma cell strain Anti-Em-11 disclosed by the invention is CGMCC (China General Microbiological Culture Collection Center) No. 46598. According to the present invention, the specific monoclonal antibody is successfully prepared and identified, and Western blot and indirect immunofluorescence experiments prove that the obtained monoclonal antibody has high specificity on the recombinant protein. The invention lays a solid foundation for serological diagnosis of the echinococcosis and remarkably enriches the existing serological detection technical system.

The invention discloses a hybridoma cell strain capable of secreting an anti-Ackermann muciniphila monoclonal antibody, the monoclonal antibody and application of the monoclonal antibody. Belongs to the technical field of biology. The invention provides a hybridoma cell strain capable of secreting an anti-Ackermann muciniphila monoclonal antibody. The secreted antibody is only combined with AKK bacteria and has no cross reaction with other common intestinal bacteria. Besides, through an indirect immunofluorescence localization experiment, the problem of how to detect the adhesion and colonization condition of Ackermann muciniphila in the colon of the mouse by using the prepared monoclonal antibody is solved, and the application range of the antibody is expanded.

The invention relates to a single-person indirect immunofluorescence detection method and a single-person indirect immunofluorescence detection system. A single-person reaction container is used as a fixing and detection platform for indirect immunofluorescence detection; an independent sample detection process is realized by adopting automatic liquid treatment and optical imaging; image features are automatically processed, and signals are quantized and output in a classified mode. Indirect immunofluorescence detection is directly optimized and designed from the aspect of a single-person reaction container, the problem of batch processing delay is solved, and indirect immunofluorescence detection is directly carried out on a single sample; the problem of limited degree of automation is solved, samples do not need to be transferred manually, optical imaging does not need to be set manually, and a full-automatic indirect immunofluorescence detection system is realized; the problems of sample confusion and low flexibility are solved, batch glass slides do not need to be repeatedly utilized, and emergency samples can be quickly inserted into a detection queue; the device solves the problem of low expandability, can be cascaded with a traditional analysis instrument, can be used in parallel connection with a plurality of single-thread detection devices, and is suitable for laboratories of various scales.

The invention relates to the technical field of antibodies, in particular to a 3D protein monoclonal antibody and application thereof. The monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises the following three complementary determining regions CDR: CDR1 shown as SEQ ID No.5, CDR2 shown as SEQ ID No.6, and CDR3 shown as SEQ ID No.7; the heavy chain variable region comprises the following three complementary determining regions: CDR1 shown as SEQ ID No.5, CDR2 shown as SEQ ID No.6, and CDR3 shown as SEQ ID No.7; the light chain variable region comprises the following three complementary determining regions CDR: CDR1 as shown in SEQ ID No.8, CDR2 with the sequence of WAS, and CDR3 as shown in SEQ ID No.9. The light chain variable region has the advantages that the light chain variable region can be used for preparing the light chain variable region; the invention provides a specific monoclonal antibody of a foot and mouth disease virus non-structural protein 3D protein. A Western blotting test shows that the specific monoclonal antibody specifically reacts with the non-structural protein 3D; indirect immunofluorescence tests show that the antibody has good reactivity with an FMDV O-type strain.

The application discloses a monoclonal antibody and application. A monoclonal antibody is secreted by a hybridoma cell strain 6B12G10, and an antigen epitope recognized by the monoclonal antibody is aa130-aa134 of a P30 protein of a gene I type African swine fever virus. An amino acid aa131 of the P30 protein is E. The application prepares a monoclonal antibody capable of identifying the P30 antigen of the gene I type African swine fever virus; the monoclonal antibody can directly distinguish the gene I type and the gene II type, the gene I / II recombinant type ASFV by using an indirect immunofluorescence technology; for type identification after infection of an ASFV weak strain, the application establishes a blocking ELISA method, and for the first time, effectively distinguishes whether the gene I type ASFV is infected or the gene II type, the gene I / II recombinant type ASFV is infected, effectively fills a differential diagnosis blank in the domestic and foreign aspects, and has important significance for tracing a gene I type ASFV infection.

This invention relates to the field of molecular biology, specifically to a nested PCR primer set, kit, and application for detecting porcine infectious pleuropneumonia. The kit, developed by establishing a nested PCR amplification primer set, can be used for the in vitro diagnosis of porcine infectious pleuropneumonia. It exhibits high sensitivity, detecting up to 0.355 pg / µL, which is nearly 100 times higher than that of conventional PCR (35.5 pg / µL). It also demonstrates strong specificity, overcoming the problems of low sensitivity and poor specificity associated with conventional PCR. Compared to existing indirect immunofluorescence and serological detection methods, it offers lower cost and a shorter detection cycle.

The invention discloses an application of licochalcone B (Lcochalcone B, LCB) in a medicine for preventing and treating porcine epidemic diarrhea, and belongs to the technical field of medicines. The antiviral effect of the licochalcone B on the porcine epidemic diarrhea virus (PEDV) at the level of Vero cells (African green monkey kidney cells) is researched through methods such as an indirect immunofluorescence experiment (IFA) and real-time fluorescent quantitative PCR (qRT-PCR), and experimental results show that the licochalcone B can remarkably inhibit proliferation of the porcine epidemic diarrhea virus. It is proved that licochalcone B has remarkable porcine epidemic diarrhea virus resisting activity, a brand-new effective candidate medicine is provided for prevention and treatment of porcine epidemic diarrhea, and the licochalcone B has good application prospects in the field of medicine for preventing and treating porcine epidemic diarrhea.

The application discloses a recombinant pseudorabies virus strain expressing classical swine fever virus (CSFV) recombinant E2 protein and application, and relates to the technical field of biology.The TK / gE double gene deletion PRV attenuated strain is used as a virus carrier, an exogenous CSFV E2 gene with a transmembrane region removed is inserted after a PRV gC signal peptide sequence, and a recombinant virus rPRV-delTK / gE-gC-E2 for expressing the gC N terminal and fusing with the PRV gC is constructed; Western blotting analysis and indirect immunofluorescence detection prove that the gC-E2 protein is successfully expressed.The recombinant virus immunized mice can produce specific antibodies against E2; after immunizing piglets, the piglets can produce an antibody level equivalent to that of a classical CSFV attenuated vaccine strain C strain.The rPRV-delTK / gE-gC-E2 obtained in the research is a promising bivalent live vaccine candidate strain.

This invention utilizes a machine learning model to analyze the crystal structure of the PEDV Nsp5 protein. Through virtual peptide screening, a peptide sequence KYRRQH (177080) with potential affinity for the target protein was identified. The 177080 sequence was artificially synthesized in a solid phase, and the peptide was screened using ELISA with artificially expressed PEDV Nsp5 protein. The affinity constant between the peptide and the target protein was determined using surface plasmon resonance (SPR) assays. The cytotoxicity of peptide 177080 was tested using a CCK-8 assay kit. Its inhibitory activity against viral infection was assessed using quantitative real-time PCR and indirect immunofluorescence assays. The results showed that 177080 significantly inhibited PEDV infection.