50 results about "Conserved sequence" patented technology

The invention discloses a method and a device for identifying conservative non-coding elements (CNEs) in a genome. The method comprises the following steps: respectively comparing other genomes in a plurality of high-quality genomes with a reference genome to obtain genome comparison results; identifying the comparison result of the species evolutionary relationship topological structure and the genome to obtain a conserved sequence position information gff file; the gff file is filtered, and a CNEs.gff file is obtained; a CNEs sequence is extracted from the CNEs.gff file; determining a CNEs differentiation sequence according to the CNEs sequence; and performing function enrichment analysis on the target gene of the CNEs differentiation sequence to obtain a plurality of identification results of the CNEs in the high-quality genome. According to the method and the device, the technical problem that identification of the CNEs is incomplete or misjudged due to the fact that identification of the CNEs is often influenced by genome structure variation in the prior art is solved.

The invention provides a primer for detecting a novel bunyavirus by an RPA-lateral flow nucleic acid chromatography technology, which comprises a specific SFTSV primer pair marked by a nucleic acid sequence, namely SFTSV-F and SFTSV-R. The invention further provides a kit for detecting the novel bunyavirus by the RPA-lateral flow nucleic acid chromatography technology. The invention also provides a probe which comprises a gold nanoparticle capture probe, an SFTSV capture probe and a quality control capture probe. Meanwhile, the invention further provides application of the primer and the probe in preparation of SFTSV detection products, a specific detection kit and a specific detection method. The specific primer and the functional differentiation probe are designed aiming at a novel bunyavirus highly-conserved sequence, and an RPA isothermal amplification technology and a lateral flow nucleic acid chromatography visual detection technology are integrated, so that the rapid, sensitive and specific detection of the SFTSV is realized, the dependence on a large instrument is not needed, the detection cost and the operation threshold are reduced, and the detection efficiency is improved. And the application requirements of on-site rapid detection, large-scale screening and resource-limited regions are met.

This invention provides a method for identifying abamectin resistance genes in the fall armyworm, dsRNA, and their applications, belonging to the field of molecular biology. This invention combines genome resequencing and transcriptome sequencing to identify key abamectin resistance genes in the fall armyworm. Based on the conserved mRNA sequences of the key resistance genes GSTD6 and CTP6A18, dsRNA was designed, and RNAi technology can effectively reduce the fall armyworm's resistance to abamectin. This has significant practical implications for developing effective fall armyworm resistance management strategies to delay the emergence of resistance and for identifying new targets for pest control.

The invention provides a target, a primer and a crRNA composition for a bovine rotavirus RT-RAA-CRISPR / Cas12a detection method, and belongs to the technical field of nucleic acid visual detection. A primer and a cRNA composition are designed on the basis of a conserved sequence in BRV VP6, an RT-RAA technology is combined with a CRISPR / Cas12a system, precise variable temperature equipment is not needed, effective amplification of a BRV target gene can be obtained only under the constant temperature condition, and the problems that an existing BRV detection method needs to be improved in sensitivity, the requirements for the operation environment and precise equipment are high, and the detection cost is low are solved. And rapid detection of the BRV cannot be realized in an epidemic disease site or a basic laboratory.

This invention belongs to the field of bioengineering technology and discloses a method for specific targeting and scarless simultaneous elimination blue KPC‑2 Gene and IncL plasmid sgRNA sequences, CRISPR / Cas9 vector system and applications. This system targets the sgRNA of the IncL plasmid. blue KPC‑2 The IncL-type plasmid of the gene was designed with an sgRNA that specifically recognizes the conserved sequence at its origin of replication, thus eliminating the drug resistance gene while disrupting the plasmid's replication ability. Conjugation transfer to recipient bacteria significantly improved vector delivery efficiency. An arabinose-inducible promoter was used to control Cas9 gene expression, improving system stability and editing success rate. The vector carries a sucrose-inducible self-eliminating gene, enabling controlled vector self-elimination. This system is not a simple superposition but rather constitutes a time-controlled operational flow: efficient delivery via conjugation, controlled cleavage via induction, and finally, induced self-elimination.

The invention relates to the technical field of construction of conditional gene knockout models, and discloses a construction method of a mouse Oct4 conditional gene knockout model based on quadruple sgRNA collaborative targeting, which comprises the following steps: step 1, designing four sgRNAs, namely sgRNA1, sgRNA2, sgRNA3 and sgRNA4, in which the sgRNA1 and the sgRNA2 target 5'end and 3 'end non-coding regions of a second exon of an Oct4 gene, and the sgRNA3 and the sgRNA4 target structural domain conserved sequences; step 2, preparing an electrotransfection solution, wherein the electrotransfection solution is composed of Cas9mRNA, the four sgRNAs and an electrotransfection buffer solution; step 3, carrying out electroporation treatment on the fertilized eggs of the mice; step 4, culturing the fertilized eggs subjected to the electrotransformation treatment to a blastocyst stage; and 5, carrying out genotype identification on the cultured blastocyst. According to the scheme, the sgRNA combination and embryo electrotransfection system is optimized, the mouse Oct4 conditional gene knockout model is efficiently and accurately constructed, a reliable tool is provided for regulation mechanism research of Oct4 in mouse early embryo trigerm differentiation, and the medical transformation value of Oct4 is excavated.

The invention provides a fluorescent quantitative PCR (polymerase chain reaction) detection primer, a kit and a method for a novel variable chalmavirus VP gene, and belongs to the technical field of virus detection. A specific primer is designed and synthesized according to a conserved sequence of a VP gene of the novel variant chalmavirus, a recombinant plasmid is constructed, then the recombinant plasmid is used as a standard substance to establish a standard curve, a real-time fluorescent quantitative PCR detection system aiming at the novel variant chalmavirus is constructed, and specific detection of the novel variant chalmavirus is realized. The detection method can be used for quantitatively detecting the novel variant chalmavirus in the to-be-detected sample, and has the characteristics of high speed, high sensitivity, strong specificity and the like; the method provides a key technical support for molecular epidemiological investigation of the Sepamavirus in animal populations, is helpful for realizing early screening of infected individuals and timely blocking of virus transmission, and has important practical significance for guaranteeing population health and maintaining high mountain ecosystem stability.

The invention provides a real-time fluorescent quantitative PCR (polymerase chain reaction) detection primer, kit and method for novel variant bocavirus, and belongs to the technical field of virus nucleic acid detection. A specific primer is designed and synthesized according to a conserved sequence of a novel variant bocavirus (BoV) VP gene, a recombinant plasmid pcDNA3.1 HA-BoV-VP is constructed, then the recombinant plasmid is used as a standard substance to establish a standard curve, a real-time fluorescent quantitative PCR detection system for the novel variant bocavirus is constructed, and specific detection of the novel variant bocavirus is realized. The detection method can realize quantitative detection of bocavirus in a to-be-detected sample, has the characteristics of high speed, high sensitivity, strong specificity and the like, plays an important role in protection of endangered wild animals such as disc sheep, and has good practicability.

The invention discloses a primer combination for detecting Ralstonia pickettii and a kit for detecting the Ralstonia pickettii, wherein the kit comprises the primer combination. The primer probe provided by the invention is targeted to a specific nucleotide sequence of the Ralstonia piceatrii instead of a horizontal conserved sequence, so that the Ralstonia piceatrii and other strains in the Ralstonia can be effectively distinguished, the technical defects that an existing detection method is insufficient in specificity and easy to generate false positive are overcome, and the accuracy of a detection result is improved. The detection limit can reach 100 CFU / mL, and compared with the prior art, the detection limit is reduced, and the detection sensitivity is improved. The kit disclosed by the invention can be applied to pollution detection of medical supplies such as pharmaceutical water, dialysis water, endoscope cleaning water, blood products, sterile normal saline and the like and rapid screening of hospital infection, is strong in specificity and high in accuracy, and has relatively high popularization and application values.

The invention discloses a rift valley fever virus nucleic acid RT-RAA-CRISPR / Cas12a detection primer probe combination, a kit and a method, and belongs to the technical field of molecular detection. In order to solve the problems that an existing RVFV detection technology is high in equipment dependence, low in sensitivity, not suitable for primary laboratories and epidemic disease sites and the like in clinical application, a highly conserved sequence of an S gene of an RVFV ZH-548 strain is selected as a detection target through comparative analysis of an RVFV S gene sequence, and the RVFV ZH-548 strain S gene is detected on the basis of the target. A group of primers and probes for RVFV nucleic acid RT-RAA-CRISPR / Cas12a visual detection are researched and developed, a corresponding detection method is established, and a matched detection kit is provided. The detection method provided by the invention is high in sensitivity, strong in specificity, simple and convenient to operate and high in safety, and has remarkable advantages in scenes such as cross-border animal quarantine, epidemic area field emergency detection and grassroots pathogen rapid screening.

The invention provides a ZmSWEET4 gene related to a corn smut disease and application of the ZmSWEET4 gene in regulation and control of the corn smut disease, and belongs to the technical field of gene engineering. The invention provides a ZmSWEET4 gene capable of regulating and controlling the resistance of corn to the tumor smut disease. A coded protein contains conserved sequences as shown in SEQ ID No.1 and SEQ ID No.2. The invention also provides a preparation method of the ZmSWEET4 gene capable of regulating and controlling the resistance of corn to the tumor smut disease. According to the present invention, the ZmSWEET4a gene and the ZmSWEET4b gene in the receptor corn are respectively edited to obtain the transgenic plant in which the ZmSWEET4a gene and the ZmSWEET4b gene are respectively knocked out, and compared with the wild type corn, the gene knockout plant has characteristics of more disease resistance. The invention proves that the ZmSWEET4 gene is related to the resistance of corn to Ustilago esculenta for the first time, and can be used for creating new germplasm resistant to the corn Ustilago esculenta and developing or screening medicines capable of preventing and / or treating the corn Ustilago esculenta.

The invention belongs to the technical field of biological genetic engineering, and discloses a CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / associated protein 9) system for specific targeted traceless elimination of a blaNDM-1 gene and an IncN plasmid and application of the CRISPR / Cas9 system. According to the system, sgRNA of an original point conserved sequence is copied by designing an IncN type plasmid carrying a blaNDM-1 gene in a targeting manner, so that the replication capability of the plasmid is destroyed while a drug-resistant gene is eliminated; the carrier delivery efficiency is greatly improved by conjugational transfer to recipient bacteria; the expression of the Cas9 gene is controlled by adopting an arabinose inducible promoter, so that the constitutive toxicity of Cas9 protein is reduced, and the system stability and the editing success rate are improved; the sucrose-induced carrier self-elimination gene carried by the carrier realizes controllable self-elimination of the carrier. The system is not simply superposed, but forms an operation process with a controllable time sequence: efficient delivery is firstly carried out through joint, then cutting is controlled through induction, and finally self-elimination is induced.

The present application relates to the technical field of plant gene function, and aims to provide a construction method of a homologous silencing system of a Paeonia lactiflora bud dormancy gene, comprising the following steps: designing amplification primers according to gene annotation and a conserved sequence region in a Paeonia lactiflora bud dormancy full-length transcriptome, and constructing a conserved sequence fragment of a target gene into Agrobacterium tumefaciens containing a pTRV vector; taking a rhizome of a treated Paeonia lactiflora annual seedling as an infection material, and placing the infection material in Agrobacterium tumefaciens liquid containing pTRV1, pTRV2 and pTRV2-target gene fragments to perform infection treatment; potting the rhizome and moving the rhizome into a culture room to perform culture, observing phenotype and expression amount differences between the rhizome and a control group in a growth process, and confirming that the homologous silencing system is successfully constructed. The present application first completes miniaturization treatment and batch bacterial liquid infection of an underground rhizome system of Paeonia lactiflora, establishes a virus-mediated homologous function silencing system of a Paeonia lactiflora bud dormancy gene, and realizes the purpose of verifying functions of bud dormancy and even germination and growth related genes.

The invention discloses a recombinant protein as well as a coding gene and application thereof, and belongs to the technical field of bioengineering. The invention designs a recombinant protein which is composed of two subunits with different functions, and design and transformation are carried out on the basis of a conserved sequence of an alpha herpes virus protein with an immune escape function, so that the secretory expression of the recombinant protein is promoted, and the defects of inclusion body formation, overhigh host toxicity and high target protein purification difficulty are overcome. Experimental results show that the recombinant protein provided by the invention has a remarkable effect of inhibiting innate immunity of cells, and can effectively inhibit interferon generated by innate immunity of cells, so that the titer of cultured viruses is improved.

The invention relates to the field of biological detection, in particular to a primer, a probe, a detection method and a kit aiming at chikungunya virus. The primer pair and the probe take the conserved sequence of the chikungunya virus nsP1 gene as a target, are suitable for detecting the chikungunya virus through an RT-RPA-LFD method, and can realize accurate, rapid and efficient detection of the chikungunya virus.

The invention discloses a quadruple fluorescent quantitative PCR (Polymerase Chain Reaction) detection method for simultaneously detecting multiple pathogens in bovine red blood cells. The method comprises the following steps: acquiring core functional gene sequences of four pathogens; carrying out gene sequence comparison, and screening out continuous conservative fragments; comparing the conserved sequence with the bovine genome and the homology of the microorganisms to obtain a specific conserved sequence; designing four pairs of PCR primers with the same annealing temperature, and designing a probe; selecting a gene sequence fragment, treating the gene sequence fragment with restriction enzyme, and connecting a target gene fragment with a T vector by using DNA (Deoxyribose Nucleic Acid) ligase; transforming the connection product into competent cells for culture, determining successful cloning, and extracting plasmids; the PCR primer and the probe are tested for multiple times, and the annealing temperature and the component concentration of a reaction system are adjusted. By implementing the method disclosed by the invention, babesia, theileria, anaplasma and eperythrozoon can be detected at the same time, the diagnosis efficiency is remarkably improved, and the risk of missed diagnosis is reduced.

The application belongs to the field of biological detection, and particularly relates to an isothermal amplification kit for detecting porcine circovirus type 3. The application provides an isothermal amplification detection primer group for porcine circovirus type 3 (PCV3), which comprises a pair of outer primers (LOF and LOR) and a pair of inner primers (LIF and LIR) and is designed according to the conserved sequence of the porcine circovirus type 3. The primer group has low design difficulty, and maintains similar sensitivity and specificity to other isothermal amplification reactions such as a loop-mediated isothermal amplification method, can be suitable for target sequences with higher GC content and below 200 bp, has no denaturation and annealing process, is suitable for on-site detection, and has great application potential.

The invention provides a detection method for detecting Sf9 host DNA residues by using a fluorescent probe and application, and belongs to the technical field of biological detection.According to the method, a specific primer pair (such as SEQIDNO: 1 / 2) and the fluorescent probe (such as SEQIDNO: 11) are designed according to a high-copy conserved sequence of Sf9 host DNA, a kit containing a gene amplification solution, a standard substance and a quality control substance is matched, DNA is extracted through a liquid nitrogen grinding-column extraction method, and the detection result is accurate. And carrying out real-time fluorescent quantitative PCR amplification and then carrying out detection. According to the method, the detection sensitivity reaches 0.003 pg / mu L, the linear range is 300 pg / mu L-0. 003 pg / mu L, cross reaction to irrelevant DNA is avoided, the repeatability CV% is smaller than or equal to 10%, the reagent is still stable after being frozen and thawed for 10 times, and the whole process is completed within 1.5 hours; the method solves the problems of insufficient specificity and sensitivity, tedious operation and the like of the existing method, and can be used for quality control of biological products such as gene therapy drugs and cell vaccines.

The invention discloses a multiple qPCR (quantitative polymerase chain reaction) detection composition, method and kit for detecting various DNA / RNA (deoxyribonucleic acid / ribonucleic acid) viruses, and aims to detect various human DNA / RNA viruses possibly existing in biological products, compare and analyze through database analysis, find virus genome conserved sequences, design primers and probes with higher specificity and detect most subtypes of the viruses. A plurality of pairs of probe primers are designed for a virus conserved region, the primer probes with good specificity and high sensitivity are finally screened, the virus content of 2 * 10 < 7 > copies / [mu] L to 20 copies / [mu] L can be accurately quantified, and the sensitivity of most viruses can reach 2-10 copies / [mu] L. By repeatedly screening and optimizing a reaction system, including the concentrations of the primers and the probes, the detection method can be used for stably detecting various DNA / RNA viruses under the condition of fewer nucleic acid samples, is simple to operate and has higher sensitivity.

The application provides a method for sequencing small circular DNA molecules, comprising: constructing recombinant transposon fragments; enriching small circular DNA molecules; constructing recombinant small circular DNA molecules; and verifying positive clone vectors. Antibiotic resistance genes are randomly inserted into each small circular DNA molecule by using transposons, so that at least one antibiotic resistance gene is obtained in each recombinant small circular DNA molecule. Since an antibiotic is added to the culture medium in the subsequent culture process, only the clone vectors successfully transformed with the recombinant small circular DNA molecules can survive and reproduce in the culture medium, so that the antibiotic resistance gene becomes a "conserved sequence" of the recombinant small circular DNA molecule, and a corresponding PCR amplification can be designed for the antibiotic resistance gene. In addition, the antibiotic resistance gene sequence can be removed by using software when analyzing the sequence of the small circular DNA molecule.

The application discloses a triplex forming oligonucleotide fluorescent probe and a detection method for detecting mycoplasma by targeting P1 adhesion protein genes, and belongs to the technical field of microbial molecular detection. The fluorescent probe is a triplex forming oligonucleotide (TFO), targets a specific conservative sequence of mycoplasma P1 adhesion protein genes, the 5' end of the fluorescent probe is modified with a fluorescent reporter group, the 3' end of the fluorescent probe is modified with a fluorescent quenching group, the fluorescent probe can form a stable triplex DNA complex with the target sequence through Hoogsteen base pairing and generate a fluorescent signal. The detection method comprises four core steps of sample pretreatment and DNA extraction, fluorescent probe incubation, fluorescent signal detection and result judgment, and does not need PCR amplification, and is simple in operation and rapid in detection. The probe has high specificity and high sensitivity, the detection lower limit can reach 1 copy / muL, is suitable for mycoplasma detection of various types of samples, and can be widely applied to scenes such as clinical diagnosis, biological pharmaceutical quality control and livestock breeding quarantine.

The application belongs to the field of agricultural microorganism application, and particularly relates to a primer group, a reagent or a kit, and a strain screening method and application adopting the same. The primer group is designed according to a conserved sequence of a structural gene fragment in an iturin A synthesis gene cluster. The structural gene in the iturin A synthesis gene cluster includes: ituA gene, ituB gene, and ituC gene. The primer group includes: primer pair I, primer pair II, and primer pair III. The primer group of the application is designed according to a conserved sequence of a structural gene fragment in an iturin A synthesis gene cluster, so as to specifically amplify a key structural gene ( ituA gene, ituB gene, and ituC gene) in the iturin A synthesis gene cluster. By using the primer group, combining a PCR amplification technology and an electrophoresis detection technology, and then rapidly, accurately and low-costly screening a bacillus positive strain capable of producing iturin A, the application has a wide application prospect.​​​​​​​​​​​​

The invention provides an H1N1 broad-spectrum mRNA vaccine with HA and NA proteins connected in series as a target based on key amino acid mutation and a preparation method of the H1N1 broad-spectrum mRNA vaccine, and relates to the technical field of vaccine preparation. The amino acid sequence of the mRNA vaccine is as shown in SEQ ID NO.1, and the nucleotide sequence of the mRNA vaccine is as shown in SEQ ID NO.2, namely on the basis of an HA conserved sequence, variation sites with amino acid property crossing near a main antigen site of an HA head and at a receptor binding part are reserved, an amino acid sequence containing a key mutation site HA'is obtained, and GGGSGGSGGSGGGGS is adopted to connect HA 'and NA conserved amino acid sequences, so that the mRNA vaccine is obtained. The defects in the prior art are overcome, the broad-spectrum protection effect on the H1N1 type influenza virus is improved by reserving the key amino acid mutation site in the HA conserved sequence, the vaccine production cost is reduced, the immune cycle is shortened, and an effective coping scheme is provided for the influenza outbreak season.

This invention discloses a PSR primer, kit, and detection method for identifying Pediococcus lactis based on a specific gene target. Based on the conserved sequence of the target gene, this invention designs and optimizes PSR-specific primers, establishes a PSR amplification system, and further provides a kit containing the primers and its application method. The PSR detection method established by this invention features high sensitivity, strong specificity, rapid reaction, simple operation, and low cost. It is suitable for the rapid and accurate identification of Pediococcus lactis and has promising application prospects in food fermentation, probiotic detection, and microbial quality control.

The invention discloses a primer group for identifying and distinguishing mycobacterium tuberculosis and nontuberculous mycobacterium, the primer group is a pair of RPA (recombinase polymerase amplification) primers for specifically amplifying a target gene of the mycobacterium tuberculosis, and the target gene is a conserved sequence rpoB gene of the mycobacterium tuberculosis. Accordingly, the inventor also develops a corresponding composition and a kit. On the basis, a method for identifying and distinguishing mycobacterium tuberculosis and nontuberculous mycobacterium based on RPA-CRISPERCas12a for a non-diagnostic purpose is established, firstly, a sample to be detected is subjected to RPA amplification, and the primer group is adopted for RPA amplification; and then, carrying out CRISPR / Cas12a detection on the RPA amplification product by adopting the crRNA and the CRISPR / Cas12a detection reagent. The method has the advantages of low cost, high practicability and simplicity and convenience in operation, is suitable for quickly screening or identifying and distinguishing the mycobacterium tuberculosis and the four non-tuberculosis mycobacteria on site, and has a wide application prospect.

The invention discloses an Eucommia virus induced gene silencing system as well as a construction method and application thereof. The system is obtained by infecting Eucommia ulmoides with a target gene fragment recombinant virus plasmid carrying a target gene through agrobacterium tumefaciens; a target gene is an Eucommia ulmoides phytoene dehydrogenase gene, namely an Eucommia ulmoides EuPDS gene, and the nucleotide sequence of the EuPDS gene is as shown in SEQ ID NO. 1; the target gene segment is a conservative CDS sequence on the EuPDS gene of Eucommia ulmoides, and the nucleotide sequence of the target gene segment is as shown in SEQ ID NO.2. The Eucommia virus-induced gene silencing system can be used for studying Eucommia EuPDS gene functions, Eucommia phytoene dehydrogenase metabolic pathways or Eucommia stress resistance, and has profound significance for studying the Eucommia gene functions.

The invention discloses a digital PCR quantitative analysis clonal IG rearrangement detection method, and relates to the technical field of molecular diagnosis, and the detection method comprises the following specific steps: extracting genome DNA or cfDNA from a sample, and storing for later use; designing a specific primer and a probe aiming at an IG VDJ conserved sequence; carrying out primer verification, and carrying out sequencing verification after electrophoresis detection; after the primer verification is passed, digital PCR detection is carried out to ensure that the contrast meets the requirement; finally, increasing the loading amount of the reinspection sample, forming a repeated hole, repeatedly detecting and summarizing data; specific primer design and a digital PCR technology are combined, a forward primer is customized, a universal reverse primer and a probe are designed, the reliability is guaranteed through strict primer verification, a contrast is set to improve the sensitivity, meanwhile, a whole-process standardized system is established, parameters of all links are finely controlled, risks are reduced, and accurate and stable detection is provided for clinic.

The invention discloses a qPCR (quantitative polymerase chain reaction) method for quantitatively analyzing human cells in a non-humanized animal model, which comprises the following steps: taking a non-humanized animal model sample, and extracting genome DNA (deoxyribonucleic acid); the method comprises the following steps: designing and synthesizing a specific primer probe combination aiming at a human COX1 gene conserved sequence; the method comprises the following steps: cloning a specific fragment of a human COX1 gene to a plasmid vector, and preparing a linearized plasmid standard substance; carrying out 10-fold gradient dilution on the linearized plasmid standard substance to obtain a standard substance gradient solution, and fitting a linear regression curve; taking the extracted DNA as a template, adding a primer probe combination and a qPCR premixed solution, and carrying out qPCR amplification; and calculating the human cell content in the sample according to the linear regression curve. The method provided by the invention has the advantages of stable standard substance, strong species specificity, high detection sensitivity, cross-species universality and simple operation, can effectively avoid animal matrix DNA interference, solves the problems of quantitative deviation and low-abundance cell detection, and reduces the detection cost.

The application is designed and screened according to the specific amplification primer of the conserved sequence of Chinese mitten crab microsporidium, the specific amplification primer screened is modified with a marker, and then the MIRA reaction condition is optimized; the MIRA detection technology has obvious advantages such as no need for thermal cycling, short detection time, simple operation, high sensitivity, simple equipment configuration and the like; compared with ordinary PCR detection, the MIRA detection technology of Chinese mitten crab microsporidium established in the experiment can complete the reaction at 37 DEG C in only 25 min, the minimum detection limit is 1 copies / µL, which is 10 4 copies / µL higher than that of ordinary PCR, the detection accuracy reaches 100%, and good specificity is achieved.

The application discloses a fluorescent probe primer, a kit and application of the fluorescent probe primer for five bodies of pigs, wherein the five bodies of pigs include porcine rickettsia, porcine chlamydia, mycoplasma hyopneumoniae, toxoplasma and leptospira. The application selects the conserved sequences of porcine rickettsia ORF5 genes, porcine chlamydia glgp genes, mycoplasma hyopneumoniae P95 genes, toxoplasma B1 genes and leptospira CLPS genes to design probe primers, the kit can simultaneously analyze a large number of samples, and whether the samples contain mycoplasma hyopneumoniae, chlamydia, rickettsia, toxoplasma and leptospira can be identified at one time, and the kit has the characteristics of simplicity, rapidness, good stability, high sensitivity and strong specificity.