74 results about "P. multocida" patented technology

The invention discloses a pasteurella multocida phage, which is characterized in that the pasteurella multocida phage is GXMP-H1 (Pasteurella multocida phage GXMP-H1), the preservation number of the pasteurella multocida phage is CCTCC (China Center for Type Culture Collection) NO: M 20242966, and the preservation number of the pasteurella multocida phage is CCTCC NO: M 20242966. The pasteurella multocida bacteriophage is applied to preparation of drugs for preventing and treating pasteurella multocida infection diseases. The pasteurella multocida bacteriophage GXMP-H1 disclosed by the invention can be used for effectively cracking five swine pasteurella multocida strains; the pasteurella multocida phage GXMP-H1 can continuously inhibit bacteria for a long time, is high in stability and good in safety, can be easily prepared into a preparation and the like, and has good killing, preventing and controlling effects on environments, feeds and the like polluted by pasteurella multocida.

The invention discloses bacillus velezensis, a microbial preparation as well as a preparation method and application of the microbial preparation, and relates to the technical field of microorganisms. The bacillus velezensis strain is preserved in the China Center for Type Culture Collection, and is named as bacillus velezensis XNW-BLSYB1, the preservation number is CCTCC NO: M 2025721, the preservation time is April 8, 2025, and the 16S rDNA sequence of the bacillus velezensis strain is as shown in SEQID No.1. The bacillus velezensis strain has the advantages that the bacillus velezensis strain is named as bacillus velezensis XNW-BLSYB1; the spore formed by the bacillus velezensis XNW-BLSYB1 strain can survive at high temperature, and can inhibit the reproductive and respiratory syndrome virus and inhibit the activity of bacteria such as streptococcus suis, infectious actinobacillus pleuropneumoniae, clostridium perfringens, pasteurella multocida and escherichia coli.

The application discloses a fusion protein for preventing and treating various pathogenic Pasteurellaceae bacterial infections, a nucleic acid vaccine and application thereof. The fusion protein comprises six antigens rplE, rpsC, rpsE, EF-Tu, DnaK and EF-G or three antigens EF-Tu, DnaK and EF-G, and the antigens are highly conservative in various Pasteurellaceae bacteria. The application further provides a recombinant nucleic acid molecule encoding the fusion protein, an immunogenic composition containing the same and a recombinant vaccine. The vaccine can induce broad-spectrum cross immune protection and is used for preventing and treating infectious diseases caused by Pasteurella multocida, Haemophilus parasuis, Mannheimia haemolytica, Haemophilus influenzae and Haemophilus paragallinarum.

The invention relates to a porcine bacterial polysaccharide-viral protein conjugate vaccine as well as a preparation method and application thereof, and belongs to the technical field of animal vaccines. The vaccine takes porcine bacterial polysaccharide and virus protein as raw materials, and a polysaccharide-virus protein conjugate is formed through coupling of a biotin-avidin system; wherein the virus protein is selected from one of classical swine fever virus E2 protein, porcine circovirus type 2 Cap protein, porcine pseudorabies virus gD protein and the like, and the porcine bacterial polysaccharide is selected from one of streptococcus suis capsular polysaccharide, porcine pasteurella multocida capsular polysaccharide and the like. The vaccine can simultaneously induce an organism to generate a high-level IgG antibody aiming at porcine bacterium capsular polysaccharide and virus protein, so that synergistic immune protection on porcine bacterium and virus infection is realized, and a broad-spectrum cross immune effect is achieved.

This invention discloses a method for preparing a bivalent inactivated vaccine against bovine Pasteurella multocida type A and B and hemolytic Mansicular disease, belonging to the field of vaccine technology. The invention involves isolating capsular serum Pasteurella multocida type A, Pasteurella multocida type B, and hemolytic Mansicular disease; separately expanding the culture of these bacteria, then inactivating and concentrating the bacterial suspensions, and resuspending them in PBS to prepare bacterial suspensions; and finally, mixing the Pasteurella multocida type A, Pasteurella multocida type B, and hemolytic Mansicular disease suspensions with aluminum hydroxide adjuvant to prepare the inactivated vaccine. This invention provides simultaneous prevention of two diseases—bovine fibrinous purulent pneumonia and bovine hemorrhagic septicemia—as well as bovine Mansicular disease through vaccine immunization. The vaccine demonstrates good safety, stable immunization efficacy, and a long duration of immunity.

The invention relates to the technical field of biology, and particularly discloses pichia pastoris as well as a construction method and application thereof. According to the pichia pastoris disclosed by the invention, beta-glucan is knocked out to synthesize a related gene gas1. By deleting the beta-glucan coding gene gas1 gene of the pichia pastoris cell wall, the cell size is reduced, the transmission distance from the cell surface to the cell nucleus is reduced, oxygen transmission and product secretion during high-density culture are improved, and the fermentation production efficiency is improved. In addition, the pichia pastoris constructed by the invention enhances the synthesis of the intracellular product bisabolol, and also can enhance the secretion ability of the intracellular product bisabolol. By reducing the cell size, reducing the carbon flow flowing to the cell wall polysaccharide synthesis and increasing the carbon source flowing to the target product synthesis, the application has great significance in industrial production.

The invention relates to the technical field of veterinary drug preparation, in particular to a bovine pasteurella multocida capsular group B disease inactivated vaccine and a preparation method thereof. The inactivated vaccine comprises an adjuvant and an antigen, wherein the antigen is a bovine pasteurella multocida capsular group B NM strain; the antigen is a bovine pasteurella multocida capsular group B NM strain which is preserved in China General Microbiological Culture Collection Center (CGMCC) on July 5, 2023, and is classified and named as pasteurella multocida with the preservation number of CGMCC No.45600. The bovine pasteurella multocida capsular group B disease inactivated vaccine provided by the invention can generate an effective and stable immune protection effect on immunized cattle when the injection dosage is 2ml. The vaccine does not need to be too high in concentration, is good in safety, does not generate abscess and nodules at an injection part, can be applied to clinic, can achieve 100% protection, is high in adhesion, and provides a good prevention effect on epidemic bovine abortion infectious diseases.

The invention provides antigen protein HPA of bovine-derived A-type pasteurella multocida and a preparation method and application of recombinant protein of the antigen protein HPA. The amino acid sequence of the antigen protein HPA is shown as SEQ ID NO.3. The preparation method comprises the following steps: cloning a coding gene sequence, shown as SEQ ID NO.1, of the antigen protein HPA after codon optimization into a prokaryotic expression vector; a recombinant plasmid containing the antigen protein HPA coding gene is obtained; transforming the recombinant plasmid into escherichia coli competent cells to obtain a recombinant expression strain; the antigen protein HPA is obtained through fermentation culture, inducible expression and purification. According to the invention, mice are immunized, the protection rate can reach 100%, and the problem of poor protection effect of the A-type pasteurella subunit vaccine for bovine pneumonia at present is solved.

The invention belongs to the technical field of biology, and particularly relates to a quintuple fluorescent PCR detection reagent for porcine respiratory pathogens and application of the quintuple fluorescent PCR detection reagent. The quintuple fluorescent PCR detection reagent disclosed by the invention can achieve the purpose of simultaneously identifying and detecting actinobacillus pleuropneumoniae, haemophilus parasuis, mycoplasma hyopneumoniae, pasteurella multocida and / or streptococcus suis, and has the advantages of simplicity, convenience and rapidness in operation, high accuracy, high sensitivity and good repeatability; the method has important significance on rapid and accurate detection and epidemic disease prevention and control of clinical porcine respiratory pathogens.

The invention belongs to the technical field of antibacterial peptides, and particularly relates to an antibacterial peptide Z34B3 and application thereof. The amino acid sequence of the antibacterial peptide Z34B3 provided by the invention is shown as SEQ ID NO: 1, the secondary structure is an alpha helix, the molecular weight is small, and the antibacterial peptide Z34B3 can change the cell membrane permeability of various gram-negative bacteria including pasteurella multocida, escherichia coli and acinetobacter baumannii, inhibit the generation of cell membranes and play antibacterial and bactericidal roles. Moreover, the antibacterial peptide Z34B3 provided by the invention has low hemolytic activity and no cytotoxicity, and can be applied to preparation of products with bacteriostatic and bactericidal effects and preparation of drugs for preventing and / or treating bacterial infectious diseases. Furthermore, the C tail end of the antibacterial peptide Z34B3 is subjected to amidation modification, and compared with non-modification treatment, the stability can be improved.

The application discloses a kit and method for visual detection of Pasteurella multocida based on CRISPR-Cas12a, wherein the kit is used for amplifying a sample to be detected, then under the mediation of a crRNA guide sequence, a CRISPR-Cas12a system is guided to recognize, combine and cut target double-strand DNA to activate a non-specific nuclease function, then an ssDNA fluorescent probe in the arbitrary cutting system is cut to obtain a cutting product and color development detection is performed to determine. The kit can detect nucleic acid of Pasteurella multocida with a sensitivity of 2 copies per muL, and can select RPA for amplification, and the whole process is performed in a 37 DEG C environment, naked eyes can directly observe, and the dependence on laboratory large instruments is broken, and the purpose of on-site rapid visual detection of target nucleic acid is achieved.

The invention belongs to the technical field of biology, and relates to a narrow-spectrum antibacterial peptide XMK-8 and application thereof. The amino acid sequence of the narrow-spectrum antibacterial peptide XMK-8 is as shown in SEQ ID NO: 1, and the narrow-spectrum antibacterial peptide XMK-8 has no bacteriostatic activity on escherichia coli, salmonella, staphylococcus aureus and aeromonas hydrophila and only has bacteriostatic activity on pasteurella multocida and haemophilus parasuis. The antibacterial activity of the narrow-spectrum antibacterial peptide XMK-8 can still be detected after treatment with salt ions (sodium ions and potassium ions, 50-200 mmol / L), acid-base (pH 4-10) and protease K (20-100 [mu] g / mL) at the temperature of 0-100 DEG C, and the narrow-spectrum antibacterial peptide XMK-8 has low hemolytic activity on red blood cells of mice. The antibacterial peptide XMK-8 disclosed by the invention is expected to become a novel antibiotic substitute and has a good application prospect in prevention, treatment, diagnosis and identification of pasteurella multocida, haemophilus parasuis and infection thereof.

The present invention relates to a strain of Clostridium butyricum and its application, belonging to the field of microbiology. The Clostridium butyricum JBH-BD1 of the present invention is preserved in the China Center for Type Culture Collection, with the preservation number CCTCC NO: M2022151. The Clostridium butyricum of the present invention is natural, safe, acid-resistant, alkali-resistant, heat-resistant, and has strong abilities to produce protease, amylase and organic acids. At the same time, it has antibacterial effects on Escherichia coli, Salmonella, Staphylococcus aureus, Clostridium perfringens, Pasteurella multocida of porcine origin and Actinobacillus pleuropneumoniae, and can convert inorganic ferrous iron into organic ferrous iron; when used in the aquaculture industry, it can not only regulate the balance of intestinal flora, promote the growth of livestock and poultry, but also effectively improve the utilization rate of iron element, reduce its addition amount, prevent iron deficiency anemia in livestock and poultry, improve the absorption and utilization efficiency of iron by livestock and poultry, and increase the aquaculture benefits.

The invention is suitable for the technical field of veterinary biological products, and provides a pasteurella egg yolk antibody for sheep and a preparation method thereof, and the preparation method comprises the following steps: S1, preparing an immunogen; s2, immunizing the hens; s3, crude extraction of an egg yolk antibody: separating egg yolk, collecting egg yolk liquid, adding a PBS buffer solution in proportion to fully dissolve the egg yolk, and standing and centrifuging to obtain a crude extract; s4, egg yolk antibody purification: precipitating the crude body fluid with an ammonium sulfate solution with the saturation degree of 38%, collecting antibody precipitate, dissolving with a PBS buffer solution, carrying out primary dialysis, then adding an ammonium sulfate solution with the saturation degree of 28%, carrying out secondary precipitation, and finally carrying out secondary dialysis to obtain a purified egg yolk antibody solution; and S5, antibody quality detection and split charging. The scheme has the advantages of comprehensive serotype coverage and strong protective force; the original animal is used for separating strains, so that the specificity is high; and the antibody has the advantages of high purity, low side reaction rate, quick response, no drug resistance, no residue and the like, and can be applied to prevention and treatment.

This invention discloses a PKPD-guided method for treating porcine Pasteurella multocida with antibiotics. YDMS injection is used as the antibiotic, administered intramuscularly to pigs infected with Pasteurella multocida. The dosing regimen is adjusted based on in vivo PK / PD target values. The minimum inhibitory concentration (MIC) of YDMS against Pasteurella multocida is 0.06 μg / mL. PK / PD monitoring is performed using a combination of colloidal gold rapid detection and quantitative real-time PCR. This invention clarifies the three-level PK / PD target values ​​of YDMS against Pasteurella multocida and uses AUIC as the core quantitative indicator, avoiding the blindness of traditional empirical dosing. This ensures precise matching of drug concentration and infection severity, effectively reducing the risk of infection aggravation due to insufficient medication or cumulative toxicity caused by overdose, and significantly improving the consistency of treatment efficacy.

This application relates to the field of biotechnology, specifically disclosing a *Pichia pastoris* yeast, its construction method, and its applications. This application involves knocking out the *gas1* gene, which is related to β-glucan synthesis, in *Pichia pastoris*. By deleting the *gas1* gene, which encodes β-glucan in the cell wall of *Pichia pastoris*, this application reduces cell size, decreases the transfer distance from the cell surface to the nucleus, improves oxygen transfer and product secretion during high-density culture, and enhances fermentation efficiency. Furthermore, the *Pichia pastoris* yeast constructed using this application enhances the synthesis of the intracellular product bisabolol and also enhances its secretion capacity. This application, by reducing cell size and decreasing the carbon flow to cell wall polysaccharide synthesis while increasing the carbon source flow to target product synthesis, has significant implications for industrial production.

The invention relates to the technical field of animal medicines, and particularly discloses a veterinary medicine compound preparation as well as a preparation method and application thereof. The veterinary medicine compound preparation is prepared by boiling, extracting, concentrating and diluting eight Tibetan medicines including aconitum pendulum, fructus chebulae, radix inulae inula, benzoin, pterocephalus hookeri, lagotis lagotis, amomum tsao-ko and rhodiola rosea. The veterinary drug compound preparation provided by the invention has an antibacterial range of 8.67 mm to 23.77 mm on enterotoxic large intestine dried bacteria, salmonella enteritidis and pasteurella multocida, and has an inhibition rate of 60% to 72% on enterotoxigenic escherichia coli diarrhea, salmonella enteritidis diarrhea and pasteurella multocida diarrhea; therefore, the veterinary drug compound preparation provided by the invention has a remarkable inhibition effect on enterotoxic large intestine dry bacteria, salmonella enteritidis and pasteurella multocida.

The invention belongs to the field of vaccine microorganisms, and provides a swine pasteurella multocida attenuated strain PmA4-750, which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: M 20251644. The invention further provides a preparation method of the swine pasteurella multocida attenuated strain PmA4-750. After alternate high-temperature continuous passage, three generations of whole-gene sequencing and pathogenicity contrast tests find that a vaccine candidate strain (PmA4-750 strain) has a section of specific insertion fragment with the length of about 43kb, and 14 genetic variation sites, including 10 InDel (insertion / deletion) and 4 SNP (single nucleotide polymorphism), exist in the vaccine candidate strain. The inserted genes may generate adaptive evolution of strains in the passage process of the strains, so that the pathogenic characteristics of the strains are changed, and the toxicity is promoted to be remarkably reduced. In addition, a piglet immune challenge protection test result shows that the PmA4-750 strain has an immune effect on swine pasteurellosis caused by type A, type D or type B pasteurella multocida. In addition, the invention also discloses a swine pasteurellosis live vaccine and a preparation method thereof.

The invention relates to the technical field of vaccines, in particular to an adjuvant of a pasteurella inactivated vaccine and application of the adjuvant. According to the specific technical scheme, the adjuvant of the pasteurella inactivated vaccine comprises radix tetrastigme polysaccharide. According to the invention, the THP adjuvant is added into the Pm inactivated vaccine to immunize mice; the Pm inactivated vaccine can effectively promote humoral immune response of the Pm inactivated vaccine, promote secretion of serum Th1 type, Th2 type and Th17 type cell factors after mice are immunized with the Pm inactivated vaccine, enhance cellular immune response of the mice induced by the Pm inactivated vaccine, increase the proportion of T lymphocyte subclasses in splenic lymphocytes of the mice inoculated with the Pm inactivated vaccine, and effectively avoid infection of Pm to the mice.

The present invention belongs to the technical field of pathogen detection, and specifically relates to an ELISA antibody detection kit for bovine Pasteurella multocida capsule type A, and its preparation method and application. The gene with the sequence shown in SEQ ID NO.1 is integrated into an expression vector and transferred into a host bacterium; the host bacterium is induced to express and purified to obtain a PmA153 recombinant antigen protein with the sequence shown in SEQ ID NO.2; PmA153 is coated on a solid phase carrier, and the detection method of bovine Pasteurella multocida capsule type A antibodies in the sample can be used to detect the bovine Pasteurella multocida capsule type A antibodies, thereby achieving the diagnostic purpose. The kit provided by the present invention does not use Pasteurella multocida as an antigen source, has good biosafety and low production costs, has good specificity for the detection of bovine Pasteurella multocida capsule type A antibodies, can accurately diagnose bovine Pasteurella multocida capsule type A pneumonia, and has important clinical application value.

The invention discloses a poultry pasteurella multocida bivalent subunit vaccine as well as a preparation method and application thereof. The poultry pasteurella multocida bivalent subunit vaccine is prepared by mixing and emulsifying a protein mixed solution consisting of PpdD-C1 protein and PpdD-C2 protein and a Freund's complete adjuvant. It is found for the first time that avian pasteurella multocida IV type pili of different serotypes are only divided into two types, and the bivalent subunit vaccine prepared based on the two types of IV type pili protein can provide a good protection rate for all the serotypes of avian pasteurella multocida. Compared with the existing vaccine, the divalent subunit vaccine prepared by the invention has the advantages of good cross protection, high safety and no endotoxin, and the stress reaction on the organism is obviously reduced.

The invention discloses an ultra-high molecular weight hyaluronic acid synthetase mutant as well as a construction method and application thereof, and belongs to the technical field of genetic engineering and biosynthesis. Hyaluronic acid synthetase derived from pasteurella multocida is modified by a method for constructing a random mutation library through error-prone PCR (Polymerase Chain Reaction), and a mutant, especially the mutant E401M, capable of remarkably improving the yield and molecular weight of hyaluronic acid in escherichia coli and corynebacterium glutamicum is obtained through screening. The invention provides an efficient and feasible technical approach for industrial production of the ultra-high molecular weight hyaluronic acid raw material.

The invention relates to the technical field of molecular diagnosis and early screening of tumors, and particularly discloses a plasma circulating microbial marker for early screening of lung cancer and application of the plasma circulating microbial marker. The marker comprises the following components: staphylococcus aureus, fusobacterium nucleatum, klebsiella pneumoniae, alternaria simplex, legionella pneumophila, neisseria mucosa, Ralstonia petii, pasteurella multocida, and haemophilus parainfluenza (Haemophilus parainfluenza), and the marker comprises the following components: staphylococcus aureus, fusobacterium nucleatum, klebsiella pneumoniae, alternaria simplex, legionella pneumophila, neisseria mucosa, Ralstonia petii, pasteurella multocida, and Haemophilus parainfluenza (Haemophilus parainfluenza). According to the method, nine characteristic microbial markers are detected in combination with a low Pass WGS technology, a high-precision diagnosis model is constructed through a machine learning algorithm, the established diagnosis model has excellent stability and clinical applicability, efficient recognition of early lung cancer can be achieved, and the detection cost can be greatly reduced.

The invention discloses a recombinant Pasteurella multocida-sourced uridine diphosphate glucuronide transferase (PmHS2) mutant and a preparation method thereof, and relates to the technical field of biology, the mutant is obtained by cloning a coding gene to a pET26b (+) vector, transforming BL21 (DE3) Escherichia coli competent cells, and then transforming the recombinant Pasteurella multocida-sourced uridine diphosphate glucuronide transferase (PmHS2) to a pET26b (+) vector. And carrying out IPTG (isopropyl-beta-d-thiogalactoside) induced culture, ultrasonication and nickel affinity chromatography purification to obtain high-purity protein. Compared with wild type recombinant PmHS2, the activity of N-acetyl glucosamino transferase of the mutant is improved by 70%, the residual activity is 67.1% after the mutant is stored at room temperature for 5 days, the soluble expression quantity is improved by 6 times, and the mutant and the preparation method have the advantages of high catalytic efficiency, high stability and high solubility, and the mutant and the preparation method are mature in process, controllable in cost and suitable for industrial production. The method can be efficiently used in the fields of heparin and derivative synthesis, traditional Chinese medicine active ingredient glycosylation and the like, is suitable for large-scale production and has a wide application prospect.

The present invention discloses a bivalent subunit vaccine for porcine contagious pleuropneumonia and porcine pasteurellosis, belonging to the field of veterinary vaccines. The bivalent subunit vaccine of the present invention contains truncated porcine Actinobacillus pleuropneumoniae antigenic proteins ApxⅠAN, ApxⅡAN, ApxⅢAN and OmpD and porcine Pasteurella multocida antigenic proteins PlpE and VacJ, which can provide cross-protection to porcine contagious Actinobacillus pleuropneumoniae and porcine Pasteurella multocida of different serotypes. The truncated proteins selected in the antigen protein combination of the present invention can all be efficiently soluble and expressed, and are easy to mass produce with low cost. The present invention provides an effective technical means for the prevention and treatment of porcine contagious pleuropneumonia and porcine pasteurellosis.

The present invention belongs to the field of genetic engineering technology and specifically relates to an attenuated Pasteurella multocida strain, a method for its construction, and applications. The attenuated Pasteurella multocida strain lacks the arcA and gatA genes of Pasteurella multocida. The arcA gene of Pasteurella multocida is numbered A0R64_00400. The arcA and gatA double-gene mutant strain constructed by the present invention is highly attenuated in ducks and has a certain level of in vivo colonization, demonstrating its potential as an attenuated live vaccine. Oral immunization with this mutant strain does not affect the production performance of ducks and can provide immune protection against a lethal dose of Pasteurella multocida. The present invention provides a new strategy for developing a live attenuated vaccine against Pasteurella multocida or other bacteria.

The application discloses a primer combination and a kit for detecting trehalose Bipartite Pasteurella, Pasteurella multocida and hemolytic Mannheimia, and belongs to the technical field of pathogenic microorganism detection. The application aims to improve the sensitivity and specificity of detection of trehalose Bipartite Pasteurella, Pasteurella multocida and hemolytic Mannheimia. The application provides a primer combination for detecting trehalose Bipartite Pasteurella, Pasteurella multocida and hemolytic Mannheimia, wherein the primer combination is primer pair 1, primer pair 2 and primer pair 3; the primer pair 1 is shown in SEQ ID NO. 1 and SEQ ID NO. 2, the primer pair 2 is shown in SEQ ID NO. 4 and SEQ ID NO. 5, and the primer pair 3 is shown in SEQ ID NO. 7 and SEQ ID NO. 8. The detection method is very stable in batch-to-batch repeatability and batch repeatability, and is suitable for large-scale clinical detection.

The invention relates to the technical field of animal medicines, and particularly discloses a veterinary medicine compound preparation as well as a preparation method and application thereof. The veterinary medicine compound preparation is prepared by boiling, extracting, concentrating and diluting eight Tibetan medicines including aconitum pendulum, fructus chebulae, radix inulae inula, benzoin, pterocephalus hookeri, lagotis lagotis, amomum tsao-ko and rhodiola rosea. The veterinary drug compound preparation provided by the invention has an antibacterial range of 8.67 mm to 23.77 mm on enterotoxic large intestine dried bacteria, salmonella enteritidis and pasteurella multocida, and has an inhibition rate of 60% to 72% on enterotoxigenic escherichia coli diarrhea, salmonella enteritidis diarrhea and pasteurella multocida diarrhea; therefore, the veterinary drug compound preparation provided by the invention has a remarkable inhibition effect on enterotoxic large intestine dry bacteria, salmonella enteritidis and pasteurella multocida.

The invention discloses a culture medium for poultry pasteurella multocida. The culture medium is prepared from the following components: 24g / L to 25g / L of tryptone, 18g / L to 20g / L of dipotassium phosphate heptahydrate, 7g / L to 8g / L of sodium chloride, 0.25 g / L of anhydrous magnesium sulfate, 1.36 g / L of monopotassium phosphate and 10g / L of glucose. The prepared culture medium for the poultry pasteurella multocida is applied to enrichment culture of the poultry pasteurella multocida, the culture medium suitable for the poultry pasteurella multocida can be obtained by optimizing the components and the component proportion of the culture medium, the culture medium is applied to the enrichment culture of the poultry pasteurella multocida, the obvious enrichment effect is achieved, and the culture medium is suitable for the poultry pasteurella multocida. The cost of the culture medium is obviously lower than that of a conventional culture medium or culture scheme.

The invention belongs to the technical field of biological detection, and particularly relates to a dual MIRA detection method for type A clostridium botulinum and staphylococcus aureus for a portable reagent card incubator. The double colloidal gold type MIRA method established by the invention has good specificity on type A clostridium botulinum and staphylococcus aureus, and has no cross reaction with six strains such as pasteurella multocida, klebsiella pneumoniae and salmonella. The lowest detection limits of the method on the staphylococcus aureus and the A-type clostridium botulinum are 2.94 * 100 copies / mu L and 2.27 * 100 copies / mu L respectively. The optimized double colloidal gold type MIRA method can be successfully applied to a reagent card incubator. The detection method disclosed by the invention has the advantages of good specificity, high sensitivity, simplicity in operation, short time consumption and the like.