Fluorescent quantitative loop-mediated isothermal amplification (LAMP) detection method for bovine tuberculosis pathogene

The invention discloses a fluorescent quantitative loop-mediated isothermal amplification (LAMP) detection method for bovine tuberculosis pathogene. The method comprises the following steps of: performing LAMP amplification on an insertion sequence IS1081 of a mycobacterium composite by internal primers shown in SEQ ID No.1 and SEQ ID No.2, external primers shown in SEQ ID No.3 and SEQ ID No.4 and loop primers shown in SEQ ID No. 5 and SEQ ID No.6 and by taking blood DNA of cattle to be detected as a template, a positive standard substance as positive control and sterile water as negative control, and collecting fluorescent signals. The method is high in sensitivity and specificity and is easy and quick to operate, and results are accurate.

Primer and probe sequence for pig streptococcus 2 type ectofactor (EF) fluorescence PCR detection

Monoclonal antibody of rat anti-medroxyprogesterone acetate(MPA)

The invention provides a specific monoclonal antibody M68F9 of mouse resisting medroxyprogesterone acetate (MPA). The antibody is prepared from mouse original hybridoma cell line M68F9, which is a kind of IfG1 subclass immune globulin, and is characterized by high specificity in binding MPA, high sensitivity, great appetency.

Fluorogenetic quantitative PCR detection method for Mycobacterium bovis Taqman

The invention provides a fluorogenetic quantitative polymerase chain reaction (PCR) detection method for Mycobacterium bovis Taqman. According to high specificity of 229bp on the Mycobacterium bovis, a primer and a probe are designed, wherein the upstream primer is 5'-TGAAGTAGTAATGTGCGAGCTGAG-3'; the downstream primer is 5'-CGTTGTAGGCCACTCCAAGAG-3'; and the probe is 5'-JOE-CGCTTCTGCACGACTACGGCTTGTATGA-Eclipse-3'. The fluorogenetic quantitative PCR detection method by which the Mycobacterium bovis can be detected simply, conveniently, rapidly and accurately, is established, is favorable for distinguishing the Mycobacterium bovis and other mycobacteria, particularly for distinguishing the Mycobacterium bovis and Mycobacterium tuberculosis, and has important significance for the control and treatment of human tuberculosis and bovine tuberculosis, the clinical detection of the bovine tuberculosis and the research of various vaccines.

Method of rapidly judging survival state of mycobacterium paratuberculosis in milk and milk product by applying BLU-V PMA technology

The invention discloses a method of rapidly judging the survival state of mycobacterium paratuberculosis in milk and a milk product by applying a BLU-V PMA technology. According to an ISMAV2 genomic sequence (with the accession number of AF286339) of Map published by GenBank, a conserved sequence of the gene is selected, and a TaqMan fluorescence probe and a primer are designed; a BLU-V system, serving as a light source, is applied to a PMA technology, and a method of rapidly detecting Map live bacteria in the milk and the milk product is successfully established with the combination of the advantages of the fluorescence quantitative PCR technology. The method is simple to operate and is rapid and accurate.

Brucella outer membrane protein antibody detection ELISA (enzyme-linked immuno sorbent assay) kit

The present invention relates to the field of biotechnology, in particular relates to a Brucella outer membrane protein antibody detection ELISA (enzyme-linked immuno sorbent assay) kit. The ELISA kit uses Brucella outer membrane protein OMP28 expressed by escherichia coli as antigen for package of an enzyme label plate, Brucella reference positive serum and negative serum are respectively used as a positive control and negative control, Goat Anti-Bovine IgG labeled with HRP (horseradish peroxidase) is used as an enzyme labeled antibody, and the ELISA kit is assembled by combination of a dilution liquid, a washing liquid, a substrate solution a, a substrate solution B and a termination liquid. The Brucella outer membrane protein antibody detection ELISA kit has the advantages of being safe, sensitive, specific, simple, fast and stable, can meet inspection and monitoring epidemic prevention requirements of Brucella antibody in cattles in China.

Fluorescent PCR primer and probe sequence for detecting swine chain coccus II virulence factor MRP

Multiple fluorescence PCR detection reagent for detecting pathogenicity of streptococcus suis serotype 2 and method

Method for purifying rabbit liver eimeria stiedai oocysts and forming aggressive spores

The invention relates to a method for purifying and extracting parasites, and particularly relates to a method for purifying rabbit liver eimeria stiedai oocysts and forming aggressive spores. The method comprises the following steps: grinding white lesion node of liver infected with rabbit liver eimeria stiedai, filtering and repeatedly centrifuging to obtain precipitate, adding a potassium dichromate solution, and performing shake cultivation for 2-6 days at 25-40DEG C, to obtain purified liver eimeria stiedai oocysts; then washing to remove potassium dichromate, then grinding and repeatedly centrifuging to obtain precipitate, digesting and performing sucrose gradient centrifugation, to finally obtain aggressive crescent spores. By adopting the method, the purified oocysts of eimeria stiedae are separated and purified from the liver of the infected rabbit and the aggressive spores can be formed, so as to make early-stage research preparation for the in vitro large-scale propagation, and ...

Primer system and method for detecting and analyzing avian influenza virus

The invention provides a method for detecting fowl influenza virus and parting primer system and augmenting fowl influenza virus RNA asymmetrically. The invention designs and screens 25 pairs multiple asymmetrical RT-PCR primers, a pair general primer, 52 specific probes and 3 quality control probes according to the report fowl influenza virus total hypotype genome sequence in Genebank, wherein every pair in 25 pairs multiple asymmetrical RT-PCR primers is fit for a hypotype of fowl influenza virus, the primer system comprising 25 pairs multiple asymmetrical RT-PCR primers and a pair general primer can be used for detecting and parting fowl influenza virus, the primer system builds the method of asymmetrically augmenting fowl influenza virus RNA and detecting and parting fowl influenza virus. The invention provides strong specificity, high sensibility, which also proceeds the first step application.

Real-time fluorescence PCR primer for perkinsosis detection and probe

The invention provides a real-time fluorescence PCR primer for perkinsosis detection and a probe; the specificity of the primer aims at a guard area of ITS2 sequence in perkinsosis genome DNA, the specificity of the probe aims at a CC high content area of the primer in an amplification area, the 5'end of the probe has a fluorescence dye reporting mark and the 3'end thereof has a quenching fluorescence dye mark; the real-time fluorescence PCR primer fully utilizes the high-efficiency amplification of the fluorescence PCR technology, good nucleotide hybridization specificity and rapid sensitivity of the fluorescence detection technology; when the reaction is finished, whether a tissue sample to be detected contains the perkinsosis can be judged according to an amplification curve; in the invention, the genome area for PCR amplification is the perkinsosis ITS2 guard area, the specificity is strong, and the primer has no cross reaction with other parasitism protozoa such as bonamiasis protozoa ...

Fluorescence quantitative RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method for rabbit hemorrhagic disease virus

The invention provides a primer and a TaqMan probe for fluorescence quantitative RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection of a rabbit hemorrhagic disease virus. The invention also provides a kit comprising the primer and the probe, and a method for detecting the rabbit hemorrhagic disease virus by using the primer and the probe with one-step RT-PCR. Due to the adoption of a one-step RT-PCR detection technology, reverse transcription and quantitative PCR amplification are continuously completed in one tube, amplification and detection are performed in a totally-closedtube, the probability of pollution caused by multiple operation is reduced, operation is easy, the volume loss in the sampling process is reduced, the dependability of a result is ensured, all cDNA (complementary Deoxyribonucleic Acid) samples obtained by reverse transcription are used for amplifying, and higher sensitivity is achieved.

Method for detecting salmonella and staphylococcus aureus in food simultaneously

The invention discloses a method for detecting salmonella and staphylococcus aureus in food simultaneously. The method includes the following steps: 1), diluting a to-be-tested food sample according to GB4789 to obtain sample homogenate of 1:10, adding 50-65wt% of nitrilotriacetic acid solution into the sample homogenate according to the volume ratio of the sample homogenate to the nitrilotriacetic acid solution being (5-7):1, vibrating the mixture, adding phosphate buffer solution or 0.05% tween-20 contained phosphate buffer solution, performing centrifugal layering, and discarding an upper layer; 2), mixing a sample obtained after processing of the step 1) with immunomagnetic beads, vibrating the mixture, subjecting the mixture to standing and layering on a magnetic frame, discarding an upper layer, and washing a lower layer of the sample, wherein the immunomagnetic beads are obtained by modifying salmonella and staphylococcus aureus antibodies, which are labeled by magnetic bead coupled ...

Underwater robot wireless charging system and method based on visual positioning

The invention discloses an underwater robot wireless charging system and method based on visual positioning, and relates to the technical field of underwater robot charging. According to the technical key points, the system comprises an underwater wireless charging device and an underwater robot; wherein the underwater wireless charging device comprises a positioning Aruco two-dimensional code, a vertical positioning rod, a wireless charging transmitting module, six supporting and fixing Y-shaped frames and two permanent magnets; a frame of the underwater robot is a 'Yue'-shaped frame composed of three layers of transverse plates, a left upper side plate, a right upper side plate, a left lower side plate and a right lower side plate. A buoyancy block, an electronic bin, a propeller, a battery bin, a mechanical claw, a wireless charging receiving module and a horizontal metal positioning rod are arranged in the frame. The method is realized based on the system, and the underwater robot is ...

Establishment of rabbit in vitro mono-layer liver cell line model as well as culture and storage methods of passing rabbit in vitro mono-layer liver cell line model to continuous liver cell strain

The invention relates to an establishment method of a rabbit in vitro mono-layer liver cell line model, as well as culture and storage methods of passing the rabbit in vitro mono-layer liver cell line model to a continuous liver cell strain, belonging to the technical field of liver cell in vitro culture. The invention aims at providing a method which can be used for replacing a collection method of rabbit liver coccidium pathogeny bred by using expensive animal experiments. The rapid, convenient and economic passage of rabbit in vitro liver cells can be stably realized through the establishment of the rabbit in vitro mono-layer liver cell line model and in vitro establishment of the culture and storage methods of passing the rabbit in vitro mono-layer liver cell line model to the continuous liver cell strain so as to achieve the purpose of breeding the coccidium pathogeny in vitro in the follow-up experiments. Through exploration and research of rabbit ages, a killing manner, digestive ...

Multi-AUV (Autonomous Underwater Vehicle) cooperative positioning method and system based on node optimization selection and factor graph

The invention discloses a multi-AUV (Autonomous Underwater Vehicle) cooperative positioning method and system based on node optimization selection and a factor graph, relates to the technical field of AUV cooperative positioning, and aims to solve the problems that the dynamic change of a system topological structure cannot be coped with, the system scale is relatively large or the system communication bandwidth is limited in the prior art. Comprising the following steps: collecting dynamic topological structure information of a system at the current moment; updating information of the slave boats and neighbor master boats thereof; constructing a factor graph model, defining a slave boat state variable, main boat position information and main boat measurement information as variable nodes, and performing transmission updating on a slave boat state variable Xk by adopting a state equation function node; the Cramer-Rao lower bound CRLBk and distance measurement evaluation factor calculation ...

Real-time fluorescence PCR primer for perkinsosis detection and probe

The invention provides a real-time fluorescence PCR primer for perkinsosis detection and a probe; the specificity of the primer aims at a guard area of ITS2 sequence in perkinsosis genome DNA, the specificity of the probe aims at a CC high content area of the primer in an amplification area, the 5'end of the probe has a fluorescence dye reporting mark and the 3'end thereof has a quenching fluorescence dye mark; the real-time fluorescence PCR primer fully utilizes the high-efficiency amplification of the fluorescence PCR technology, good nucleotide hybridization specificity and rapid sensitivity of the fluorescence detection technology; when the reaction is finished, whether a tissue sample to be detected contains the perkinsosis can be judged according to an amplification curve; in the invention, the genome area for PCR amplification is the perkinsosis ITS2 guard area, the specificity is strong, and the primer has no cross reaction with other parasitism protozoa such as bonamiasis protozoa ...