115 results about "IMMUNE FLUORESCENCE" patented technology

The invention discloses a preparation method for a PCV-II Cap protein monoclonal antibody, an antibody and application. The invention adopts ultracentrifuged and purified PCV-II as an immunogen to immunize a BALB/c mouse by the conventional method, takes spleen cells of the immunized BALB/c mouse to fuse with SP2/0 cells, obtains two strains of hybridoma cells secreting the PCV2-Cap protein monoclonal antibodies by indirect ELISA screening, respectively names the two strains of hybridoma cells as 8-60 and 10-48, identifies biological characteristics of the two strains 8-60 and 10-48, and usesthe two strains 8-60 and 10-48 as the first antibodies to establish an indirect immunofluorescence diagnostic method. The result of the indirect immunofluorescence diagnostic method is basically consistent with that of the PCR diagnostic method, and the positive and negative coincidence rates are respectively 93.75 percent and 100 percent so as to provide reference for preventing and treating theporcine circovirus disease.

The invention discloses an anti-flavivirus envelope E protein monoclonal antibody and application thereof. The anti-flavivirus envelope E protein monoclonal antibody of the invention is secreted by a mouse source hybrid tumour cell strain D2-2A10G6 with the preservation number of CGMCC No. 3292. The anti-flavivirus monoclonal antibody of the invention can be specifically combined with the function epitope of flavivirus envelope E protein. The amino acid sequence of the functional epitope of envelope E protein is SEQ ID NO: 17. The monoclonal antibody of the invention is subject to screening by indirect immunofluorescent method, and indirect enzyme-linked immunization is used for evaluating the specificity and affinity when being combined with antigen. By adopting the monoclonal antibody or immunoconjugate of the invention, flavivirus infection cell can be blocked and suckling mouse can be protected from virus attack, thus achieving the effect of inhibiting virus infection. The monoclonal antibody or immunoconjugate of the invention also can be used for flavivirus envelope E protein detection.

The invention discloses a HCMVPP65 antigenemia indirect immune fluorescence method detection kit, which uses cytomegalovirus-AD169 virus strain pp65 protein as the immunogen to immune a Balb/c mouse. The spleen cells of the immunized mouse and the myeloma cells of the mouse which belongs to the same type with the immune mouse are conventionally integrated, by indirect ELISA screening and finite dilution cloning, the hybridoma cell lines of the mouse cytomegalovirus pp65 protein cloning antibody are obtained, and the characteristics of the hybridoma cell lines are identified by ELISA, immune fluorescence experiment and other methods; two monoclonal antibodies that stably secrete the pp65 protein are established successfully and named respectively as 1A6 and 4A8. A monoclonal antibody which differs from the former report and aims at the pp65 protein of the cytomegalovirus (CMV) is prepared and a method used for preparing erythrocyte fast pyrolysis is established; compared with other detection kits which belongs to the same kind, the detection kit of the invention is faster, simpler and more convenient and has higher specificity and sensitivity.

The invention discloses an immune-fluorescence test strip component for rapidly detecting C-reactive protein quantitatively, a detection card component produced by the same and a method for preparing the same. The test strip component comprises a test strip and a platinum porphyrin marked specific antibody independently packed. The test strip comprises a bottom lining, a water absorption pad, coating analysis film and a sample pad. The coating analysis film is provided with a detection line and a quality control line, a specific antibody coated by the detection line is a C-reactive protein resistance monoclonal antibody, and a specific antibody coated by the quality control line is a rabbit intravenous gamma globulin (IgG) antibody; the detection card component comprises a test strip, a card box composed of a cover plate and a back plate and a latinum porphyrin marked specific antibody independently packed. The test strip component for detecting C-reactive protein in body fluid has the advantages of being simple in operation, rapid, sensitive, good in specificity and the like, and having good clinical application prospect.

The invention discloses two monoclonal antibodies west nile virus and an relative application, wherein the monoclonal antibody is secreted from mice hybridoma cell WNV-2F5 of preservation number CGMCC No.2261 and mice hybridoma cell WNV-4B3 of preservation number CGMCC No.2262. The antigenic determinant of the monoclonal antibody is the third structural field of membrane protein E on west nile virus envelope. The monoclonal antibody is selected by indirect enzyme linked immunity method, analyzed by western blot, detected by flow cytometry, analyzed by surface plasma resonance and indirect immunity fluorescence method to completely analyze the specificity and affinity of combination with antigen, the antibody is marked by bioepiderm to build an antigen trap indirect enzyme linked immunity detection system. The inventive monoclonal antibody can be applied in various west nile virus antigen and in the test agent box preparation of west nile virus.

The invention discloses a near-infrared fluorescence detection kit for C-reactive protein and application of the near-infrared fluorescence detection kit. The near-infrared fluorescence detection kit comprises a reagent strip positioned on a back plate, wherein the reagent strip is a sample pad, a glass cellulose membrane marked with an immune fluorescent probe, a nitrocellulose membrane coated with a C-reactive protein antibody and water absorbing paper from the sampling end in sequence. The near-infrared fluorescence detection kit disclosed by the invention can quickly and accurately detect the C-reactive protein in human whole blood, serum or plasma in combination with a near-infrared fluorescence detection technology.

The invention provides an upconversion-immunochromatography test strip for detecting Escherichia coli O157:H7, and a detection method thereof. The test strip comprises a sample pad, a conjugate pad, an analysis membrane and an absorbent pad, the sample pad and the conjugate pad are glass cellulose membranes, the analysis membrane is a nitrocellulose membrane, the absorbent pad is a cellulose membrane, the analysis membrane is provided with a detection line and a quality control line, the sample pad, the conjugate pad and the absorbent pad are sequentially pasted on a PVC plate, the conjugate pad is immobilized with an upconversion luminescence nanoparticle-Escherichia coli O157:H7 monoclonal antibody IgG conjugate, the detection line on the analysis membrane is immobilized with an Escherichia coli O157:H7 antigen, and the quality control line is immobilized with MAbF1. The above upconversion nanomaterial-based immunofluorescence test strip has the advantages of low sensitivity, wide detection range, stable signal, no photobleaching, low background, sensitive signal, and accurate and reliable result.

The invention discloses a primer group, a kit and a method for detecting six mouse viruses by multiple immunity fluorescence. The primer group is designed by the aid of multiple RT-PCR (reverse transcription-polymerase chain reaction) technologies and Luminex liquid chip technologies and can simultaneously detect six high-infection viruses such as TMEV, MHV, MNV, Reo-3, MVM and MPV of mice. By the aid of the primer group, target amplification products are acquired through specific PCR, the amplification products, fluorescence encoding microspheres and streptavidin-phycoerythrin are hybridized, and different types of viruses are distinguished when a mean fluorescence intensity value is read by a Luminex detector. The method has the advantages of rapidness, high efficiency, high specificity and sensitivity, good repeatability and the like and can be applied to quality monitoring, epidemiological investigation and early warning of test mice. The primer group is good in flexibility, and varieties detecting the viruses can be increased and decreased as required on the basis.

The invention discloses a rotary table device. The rotary table device comprises a test paper clip and a first rotary table of a pushing mechanism, the test paper clip feeds test paper automatically, and the pushing mechanism pushes the test paper away from the first rotary table; a second rotary table is an annular structure coaxially arranged on the outer side of the first rotary table in a sleeving mode; a guide-track groove is formed in the second rotary table in an embedded mode, and a first opening is formed in the portion, close to the outer edge and right opposite to the upper portion of the guide-track groove, of the second rotary table; one end of the guide-track groove is communicated with a test paper outlet, a second opening is formed in the other end of the guide-track groove, and an elastic piece is arranged at the end, close to the second opening, of the guide-track groove; the test paper extends out of the test paper outlet, is firstly pushed by the pushing mechanism and slides along the guide-track groove till the test paper is blocked by the elastic piece, and an area to be detected aligns to the first opening; the test paper is subjected to second pushing of the pushing mechanism, passes over the elastic piece and gets away from the second opening. According to the rotary table device and the full-automatic immune fluorescence analyzer with the same, test paper sample adding and automation of the whole detection process are achieved, a single form of queue waiting is not needed, and the work efficiency and stability are improved.

The invention relates to monoclonal antibody of N-terminal protein of chlamydia abortus POMP18D as well as a preparation method and application thereof. The monoclonal antibody is secreted by a hybridoma cell strain 1F10D4 with the collection number of CGMCC No.4658. The invention also provides application of the monoclonal antibody in direct fluorescence immune or indirect fluorescence immune detection of chlamydia abortus. The monoclonal antibody of N-terminal protein of chlamydia abortus POMP18D has strong specificity and fluorescent characteristic, and is suitable for being used as a fluorescence antibody to establish a direct fluorescence detection method or indirect immune fluorescence detection method.

The invention discloses a colloidal gold fluorescent quantitative analysis all-in-one machine and a control method thereof. According to the all-in-one machine, through the application of a microprocessor, as well as an illumination unit, a shooting unit, an image processing unit, a laser device and a first photoelectric sensor which are connected with the microprocessor, the all-in-one machine disclosed by the invention can be used as a colloidal gold card reader and also can be used as an immunity fluoroanalyzer, so that the cost is reduced. The invention also provides a control method of the all-in-one machine, and whether a test paper card is a colloidal gold test paper card or an immunity fluorescence test paper card does not need to be distinguished by an operator in the using process, so that the experimental procedures are reduced, and the experimental efficiency is improved.

The present invention discloses a strain of a hybridoma cell line 7D7 secreting CAV-2 monoclonal antibody, monoclonal antibody secreted by the hybridoma cell line 7D7, and applications of the monoclonal antibody, wherein the preservation number of the hybridoma cell line 7D7 is CGMCC No.10877. According to the present invention, the indirect immunofluorescence results show that: the monoclonal antibody secreted by the hybridoma cell line 7D7 can specifically recognize CAV-2 virus and do not react with MDCK cells; the superposition experiment results of the prepared monoclonal antibody secreted by hybridoma cell line 7D7 and the monoclonal antibody secreted by 2C1 show that the two monoclonal antibodies are against different epitopes of the CAV-2 virus; the prepared monoclonal antibody can well react with CAV-2, and can be used for identification of clinical isolated strains; and the prepared 7D7 monoclonal antibody and the monoclonal antibody 2C1 are the monoclonal antibodies identifying the two different antigenic determinants of the CAV-2, such that the monoclonal antibody of the present invention can be used for preparation of the CAV-2 pathogenic derection immune colloidal gold test paper strip and establishment of the double antibody sandwich ELISA method.

The invention discloses a method for separating and culturing RSV (Respiratory Syncytial Virus) by using OMC (Ostiomeatal Complex)-685-1 cells, which comprises the following steps of: using the OMC-685-1 cells as a respiratory syncytial virus separating tool, observing a cytopathogenic effect by using an inverted microscope and detecting the virus by using an indirect immuno fluorescence method, and specifically comprises the following steps of: culturing the OMC-685-1 cells, infecting an OMC-685-1 cell line, observing a CPE (Cytopathic Effect) by using the inverted microscope and detecting the respiratory syncytial virus by using the indirect immuno fluorescence method. With the adoption of the separating and culturing method, whether a collected specimen contains RSV or not can be directly judged. The method has the advantages of simplicity, good repeatability, high sensitivity, short time consumption and low cost.

The invention relates to a construction method of recombinant baculovirus for expressing serum type 4 avian adenovirus fibroid protein F2. The construction method comprises the following steps: (1) constructing an FAdV4F2 gene recombinant baculovirus expression vector; (2) packaging and passing baculovirus for expressing recombinant FAdV4 protein; and (3) transfecting baculovirus genome carrying FAdV4F2 gene into Sf9 cells, culturing in an incubator at 27 DEG C for 57 days, and observing cytopathic effect; when cytopathic effect reaches 70%, collecting cell lysis buffer, centrifuging to removeprecipitate, and preserving as seed virus; passing virus, and carrying out enlarged culture as seed virus when passing to the third generation. According to the invention, a baculovirus expression system is utilized to construct a recombinant baculovirus shuttle vector containing F2 gene, the recombinant baculovirus shuttle vector is transfected into Sf9 cells to obtain recombinant baculovirus rBAF2, and expression of recombinant protein F2 is identified by virtue of indirect immunofluorescence assay (IFA) and Western blot.

The invention provides a recombinant Lactobacillus casei for expressing the infectious pancreas necrosis virus (IPNV) VP2 protein and a preparation method thereof. According to the whole gene sequence of the IPNV VP2 protein and the gene fusion characteristic of expression vector plasmid, a pair of primers are designed to perform PCR to obtain target segment of 1095bp containing the main antigen site of the IPNV VP2 protein, the IPNV VP2 protein obtained through amplification is linked to the surface expressed carrier plasmid pPG1 and enters the cell of the host strain Lactobacillus casei 393 through electrotransformation, and the Lactobacillus casei containing positive recombinant plasmid pPG1-IPNV VP2 is named pPG1-IPNV VP2/L. casei 393. The recombinant pPG1-IPNV VP2/L. casei 393 is expressed under the induction of lactose. In the invention, the Lactobacillus casei expression carrier system of the IPNV VP2 protein is constructed and the target protein of around 40 KDa containing the main antigen site of the IPNV VP2 protein is expressed, and according to the Western blot experiment and indirect immunofluorescence experiment, the expressed foreign protein is capable of reacting with the IPVN immune serum, which shows that the recombinant VP2 protein and the IPNV natural antigen have the same antigenicity.

The invention belongs to the technical field of biology. The invention relates to a polypeptide containing two dominant epitopes of canine brain natriuretic peptide (BNP). The immune effect is enhanced by coupling the polypeptide with KLH protein immune mice. The invention also provides a recombinant protein. The recombinant protein comprises two dominant epitopes of canine BNP, and in order to improve the yield of the recombinant protein in a prokaryotic expression system, escherichia coli preferred codons are adopted to convert the amino acid sequence of the recombinant protein into a corresponding nucleotide sequence, the nucleotide sequence is chemically synthesized, and a recombinant protein expression vector is constructed. The invention further relates to a phage library established by coupling polypeptide with KLH protein immune mice, a corresponding canine BNP single-chain antibody scfv sequence is obtained through panning and screening, the obtained scfv sequence is constructed into a complete mouse IgG1 antibody sequence expression vector, monoclonal antibodies are expressed through transient HEK293F cells, the monoclonal antibodies are purified, and the optimal monoclonal antibody pairing combination is determined through an immunofluorescence orthogonal experiment.

The invention discloses a monoclonal antibody for a paralichthys olivaceus resistant T cell surface marker molecule CD4-1. The monoclonal antibody is named as a hybridoma cell JF-CD4-1-2A10, the preservation number is CCTCC NO: C201879, the preservation organization is China Center For Type Culture Collection, the address is Wuhan university of Wuhan city in Hubei province, and the preservation date is March 29th, 2018, and is secreted by the hybridoma cell. An indirect enzyme-linked immunosorbent assay reaction experiment on the monoclonal antibody shows that the monoclonal antibody can be specially combined with paralichthys olivaceus CD4-1 polypeptide, indirect immune fluorescence and flow cytometry prove that the monoclonal antibody can specially identify the paralichthys olivaceus lymphatic cell surface CD4-1 molecule, and an immunoblotting result shows that the monoclonal antibody disclosed by the invention can specially identify paralichthys olivaceus CD4-1 recombinant protein and natural paralichthys olivaceus CD4-1 protein of which the molecular weight is 52 kDa. According to the monoclonal antibody disclosed by the invention, the monoclonal antibody for the paralichthys olivaceus resistant T cell surface marker molecule CD4-1 is prepared, and an important tool is provided for detecting paralichthys olivaceus CD4-1<+T> lymphatic cells.