50results about How to "Primer specificity" patented technology

The invention provides a multiple RT-PCR detection method, which can simultaneously detect whether leaves, twigs, flowers or fruits of an apple tree bring three latent viruses namely apple chlorotic leaf spot virus, apple stem grooving virus, and apple stem pitting virus, and three apple scar skin viroids namely apple scar skin viroid, apple dimple fruit viroid, and apple fruit crinkle viroid or not. The detection method has the advantages of clear detection result, convenient operation, rapid reaction, and high sensitivity, and overcomes the shortages of time consuming, tedious steps, difficulty in distinguishing the stripes, and low sensitivity in the prior art. The invention provides a rapid detection method, which is easy for promotion and application, for detecting apple viruses.

The invention discloses a duplex PCR authentication method of cordyceps sinensis original powder. The method adopts a technical scheme that cordyceps sinensis original powder genome DNA is adopted as a template; with PCR primers designed by the invention, a single-tube duplex PCR reaction is carried out, and characteristic housekeeping genes of a cordyceps sinensis strain and a host hepialus are simultaneously amplified, wherein actual amplified fragments are respectively 320bp and 136bp; and through agarose gel electrophoresis detection, product authenticity is determined. With the method provided by the invention, cordyceps sinensis original powder can be specifically identified, and sample authenticity detection can be completed simply with three steps of DNA extraction, PCR amplification, and electrophoresis detection. The operation is simple and is easy to command, and accuracy is high.

The invention discloses a bladder cancer maker set. The bladder cancer maker set comprises the following genes of HOXA9, ONECUT2, PCDH17, PENK, TWIST1, VIM and ZNF154. The invention further provides an application of the bladder cancer maker set to preparation of a reagent or a reagent kit for diagnosis of bladder cancer, and further provides a reagent composition for detecting the methylated level of the bladder cancer maker set. The bladder cancer maker set can be used for early screening of the bladder cancer, and subtype and / or prognosis forecast of the bladder cancer, so that the diagnosis and detection sensitivity, specificity and accuracy are improved. Through accurate risk assessment and forecast, the bladder cancer maker set disclosed by the invention can reduce unnecessary invasive inspection of low-risk (low-danger) patients.

The invention relates to the technical field of security detection of agricultural and sideline products, in particular to a primer for simultaneously detecting five food-borne pathogenic bacteria through a liquid chip and a detection method. The detection method comprises steps as follows: a specific primer and a gene probe for five foodborne pathogenic bacteria are designed; DNA templates of the detected five foodborne pathogenic bacteria are prepared; PCR (polymerase chain reaction) amplification is performed on the five foodborne pathogenic bacteria; coupling is performed on the gene probe for the five foodborne pathogenic bacteria and microspheres, and a microsphere probe is obtained and verified; an obtained PCR amplification product and the microsphere probe have a hybridization reaction; a fluorescence signal of a hybridized product is detected, and quantitative analysis is performed on the five foodborne pathogenic bacteria. A method capable of simultaneously detecting five common foodborne pathogenic bacteria is successfully established, the variable coefficient is within 2%, the specificity is 100%, the sensitivity can reach 100 CFU / mL, and the method is equivalent to a fluorescent PCR technology and can be applied to high-throughput rapid detection of clinic and environment specimens of foodborne diseases.

The invention discloses an eightfold PCR amplification kit and an amplification primer thereof for detecting pathogenic bacteria in food, wherein a PCR amplification reaction solution consists of the following components: 0.4umol / L-0.7umol / L of a primer, 50-100mmol / L of Tris-HCl which is 8.0-8.5 in pH value, 50-100mmol / L of potassium chloride, 25-50mmol / L of magnesium chloride, 2-10mmol / L of dithiothreitol, 10%-15% of DMSO, 2.5-5.0mmol / L of dNTP, 0.1-0.2ug / uL of LBSA, 1% of TritonX-100, 30-60mmol / L of ammonium sulfate and 5U / uL of Taq enzyme. The amplification kit and the amplification primer disclosed by the invention can be used for synchronously detecting seven pathogenic bacteria; and the amplification kit and the amplification primer are high in sensitivity and high in specificity, and are capable of improving detection speed and reducing detection cost.

The invention discloses a specific PCR (Polymerase Chain Reaction) identification method of paecilomyces hepiali powder. The technical scheme is that the method comprises the following steps: by taking a genome DNA (Deoxyribonucleic Acid) of a sample as a template, carrying out a primary single tube specific PCR reaction by adopting a specific PCR primer designed by the invention; amplifying a characteristic housekeeping gene of winter paecilomyces hepialid, wherein the actual amplification segment is 231bp; and detecting the amplification products by agarose gel electrophoresis to judge the authenticity of the sample. The method disclosed by the invention can specifically identify the paecilomyces hepiali powder, and can detect the authenticity of the sample just by three steps: DNA extraction, PCR amplification and electrophoretic detection, so that the method is simple to operate, easy to master and high in accuracy.

The invention discloses a building method of a tsh and iss double-gene coexpression strain. The method sequentially comprises the steps of double-gene clone, connection of tsh gene and iss gene with pETDuet-1 plasmids, expression strain conversion, target gene coexpression and the like. A microbial cell model based on tsh and iss is built; the biotransformation is performed; the method is simple;the process can be easily controlled; the double genes of tsh and iss are contained in the coexpression strain; the success ratio of the double-gene expression is high. The invention also provides application of the strain to prevention and treatment of avian colibacillosis. The method provided by the invention is applicable to the building of the tsh and iss double-gene coexpression strain; a gene engineering bacterium vaccine prepared from the strain is applied to the prevention and treatment of avian colibacillosis.

The invention relates to the technical field of biology, in particular to LAMP primer set for quickly detecting salmonella, a detection method and a kit. The primer set includes inner primer FIP / BIP and outer primer F3 / B3. The detection method comprises the following steps: firstly extracting genome DNA of bacterium to be detected, performing LAMP amplification with the extracted genome DNA as a formwork, then detecting an amplification product and determining whether a sample to be detected contains salmonella or not by judging whether a reaction result is positive or not. The kit comprises Bst DNA polymerase buffer solution, Bst DNA polymerase, dNTPs, MgSO4 and Betaine. The LAMP primer set for quickly detecting the salmonella has advantages of strong specificity, high flexibility, convenience to operate, strong repeatability, good practicability and the like.

The invention discloses a loop-mediated isothermal amplification detection method for detecting Fusarium oxysporum of Chinese wolfberry root rot, and detection primers and a verification method thereof. The detection primers are designed by adopting LAMP (Loop-mediated Isothermal Amplification) design software Primer Explorer V4, and comprise two outer primers F3 and B3 and two inner primers FIP and BIP. The invention has the following beneficial effect: By the detection method, the primers and the verification method, five Fusarium oxysporum from different geographical sources is yellow green, namely positive after LAMP detection, but the control and other pathogenic bacteria are orange, namely negative. The invention has the advantages of strong primer specificity, high detection sensitivity, high reaction speed, direct observation of results by naked eyes, high accuracy, difficulty in pollution, direct detection of suspected disease samples and the like. And meanwhile, comparison and judgment are performed through color development, so that the detection accuracy is greatly improved.

The invention discloses a noninvasive detection method of bladder cancer. The noninvasive detection method of bladder cancer can realize early screening of bladder cancer, risk assessment about the fact that a subject suffers from bladder cancer, and parting and / or prognostic prediction of bladder cancer, the sensitivity, specificity and accuracy of diagnosis are improved, and the effect of noninvasive, convenient and rapid, efficient and rapid diagnosis of bladder cancer is realized. The invention further provides a bladder cancer diagnosis system, a reagent combination for detecting the methylation level of a target gene in urine, and a method for detecting the methylation level of the target gene in the urine.

The invention discloses a specific PCR (Polymerase Chain Reaction) identification method of cordyceps militaris. The technical scheme of the specific PCR identification method is as follows: genome DNA (Desoxyribo-Nucleic Acid) of a sample is taken as a template; a specific PCR primer designed by the specific PCR identification method is adopted to carry out one-time single-pipe specific PCR reaction so as to amplify the characteristic housekeeping gene of the cordyceps militaris; the actual amplification segment is 417bp; then the amplification product is detected through agarose gel electrophoresis to determine the authenticity of the sample. The specific PCR identification method is capable of specifically identifying the cordyceps militaris, and also capable of completing authenticity detection on the cordyceps militaris and the products thereof in different forms such as mycelium, fungus powder and capsules only through DNA extraction, PCR amplification and electrophoresis.

The invention belongs to the technical field of plant virus detection, and provides a multiplex PCR (Polymerase Chain Reaction) detection kit for a garlic virus. The kit comprises five groups of primer pairs including SLV, GCLV, LYSV, OYDV and Allexiviruses, and the nucleotide sequences of the five groups of primer pairs are as shown in SEQ ID NO: 1-10. The garlic virus is at least one of an SLV (shallot latent virus), a GCLV (garlic common latent virus), an LYSV (leek yellow stripe virus), an OYDV (onion yellow dwarf virus) and an Allexivirus. The garlic virus detection kit provided by the invention can be used for simultaneously detecting 12 kinds of viruses infecting garlic in China, such as an SLV, a GCLV, an LYSV, an OYDV and a shallot X virus through one-time PCR reaction, so that the detection efficiency is improved, and the detection cost is reduced. According to the present invention, the used primers have characteristics of strong specificity and reasonable target band size distribution, and can effectively identify the type of the infected virus while the accuracy of the detection result is ensured.

The invention discloses an in-vitro amplification detection method for novel coronavirus nucleic acid and a test kit thereof. The method comprises the following steps: (1) extracting to-be-detected DNA; and (2) taking the to-be-detected DNA as a template, and carrying out reverse transcription NCA-QPCR by adopting primers as shown in SEQ ID NO. 1-6. The method provided by the invention can effectively solve the efficiency of specific amplification and the inhibition effect of non-specific amplification on specific amplification, and can rapidly, accurately and sensitively detect the novel coronavirus nucleic acid.

The invention discloses an application of a reference gene in preparation of a reagent or a kit for detecting expression level of a target gene in an exosome, and a reference gene combination and an application of the reference gene combination in preparation of a reagent for detecting expression level of a target gene in an exosome. The invention also provides a reagent combination and a method for detecting expression level of a target gene in an exosome. Compared to an exogenous reference gene, the reference gene of the present invention can achieve higher accuracy of detection results.

The invention relates to the technical field of plant disease pathogenic bacteria detection, in particular to a highland barley stem rot fusarium avenaceum loop-mediated isothermal amplification detecting method and application thereof. In a LAMP system, four primers specifically recognize six regions of a target gene, but in a PCR system, only two primers recognize two regions. The four primers are designed by a LAMP design software, and comprise two outer primers F3 and B3 and two inner primers FIP and BIP, highland barley stem rot fusarium avenaceum of seven different geographical origins is yellow-green positive through LAMP detection, gradient bands appear in 2% agarose gel electrophoresis detection, but a contrast group and other pathogenic bacteria are orange negative, and bands donot appear in electrophoresis detection. The four primers are designed by LAMP, and the effects can be implemented. Meanwhile, comparison judgment is carried out by two methods of electrophoresis anddeveloping, and the detection accuracy is greatly improved.

The invention discloses quintuple PCR primers for detecting pathogenic bacteria in fresh agricultural products, a probe and a detection kit. The quintuple PCR primers comprises five primer pairs, and the five primer pairs are salmonella forward primer and salmonella reverse primer, staphylococcus aureus forward primer and staphylococcus aureus reverse primer, escherichia coli O-157 forward primer amd escherichia coli O-157 reverse primer, vibrio parahaemolyticus forward primer and vibrio parahaemolyticus reverse primer, and listeria monocytogenes forward primer and listeria monocytogenes reverse primer. The sequence information is shown as SEQ ID No.1-10. According to the technical scheme, five pathogenic bacteria can be simultaneously detected, interference among the primers is small, non-specific amplification reaction is less, detection sensitivity and specificity are guaranteed, detection speed is improved and also detection cost is reduced.

The invention discloses a method for rapidly detecting the polymorphism of a sugarcane sucrose phosphate synthase B (SPSB) gene. The method comprises the steps of designing a pair of primers to carry out PCR amplification by taking the whole-genome DNA of saccharum robustum B.51NG3, saccharum species ROC22, saccharum.officinarum L.Badila, saccharum sinense R. lustrous bamboocane, saccharum barberi J.Nagori or saccharum spontaneum L.yinge No.1 as a template; and carrying out agarose gel detection to name the SPSB genotype of a fragment with the size of 175bp as type B1, name the SPSB genotype of a fragment with the size of 294bp as type B2 and name the SPSB genotype of a fragment with the size of 421bp as type B3. The invention provides a DNA level identifying method for saccharum sugar trait marker-assisted selection, and the DNA level identifying method is beneficial to tracking of different haplotypes of SPSB genes in filial generations of saccharum and has important significance for rapidly establishing saccharum germplasm with excellent heredity.

The invention relates to the technical field of agriculture, in particular to a COI primer for PCR detection of peanut aphids and application thereof. The primer comprises a forward primer HS-F1 and areverse primer HS-R1. The primer is high in specificity, can be used for amplifying a clear and single specific band only for the peanut aphids, and has no amplification effect on other species in the same environment with the peanut aphids. The invention further provides a method for detecting natural enemies of the peanut aphids through a PCR method. Therefore, a technical support is provided for qualitative and quantitative evaluation of the nutritional relationship between predators and prey, the control effect of various predatory natural enemies on the peanut aphids can be determined conveniently, the ecological regulation and control effects of the natural enemies on the peanut aphids can be enhanced, and the purposes of controlling pests and increasing the yield are achieved.

The invention discloses a method for rapidly detecting the polymorphism of a saccharum sucrose phosphate synthase B (SPSB) gene, belonging to the field of molecular breeding of plants. The method comprises the steps of designing a pair of primers to carry out PCR amplification by taking the genome DNA of saccharum robustum B.51NG3, saccharum barberi J.Nagori, saccharum species ROC22, saccharum.officinarum L.Badila, saccharum sinense R. lustrous bamboocane or saccharum spontaneum L.yinge No.1 as a template; carrying out agarose gel detection to name the SPSB genotype of a fragment with the size of 287bp as type C1 and name the SPSB genotype of a fragment with the size of 331bp as type C2; and identifying the polymorphism of the saccharum SPSB gene according to different sizes of the fragments. The invention provides a DNA level identifying method for saccharum sugar trait marker-assisted selection, and the DNA level identifying method is beneficial to tracking and utilization of different haplotypes of SPSB genes in filial generations of saccharum and has important significance for rapidly establishing saccharum germplasm with excellent heredity.

The invention relates to a primer group, a probe group and a kit containing the primer group and the probe group, which are used for synchronously detecting various common digestive tract infection related bacteria of children and can be used for simultaneously detecting various common bacteria causing digestive system infection by one-time reaction. The kit is the most comprehensive children digestive tract bacterial infection detection method at present, is high in efficiency and specificity, and provides an auxiliary diagnosis basis for clinical examination. According to the technical scheme, the technical problem that in the prior art, diagnosis and research on bacterial pathogenic bacteria related to children digestive tract infection are still insufficient can be solved.

The invention discloses primers and a kit for detecting tomato gray leaf spot pathogenic bacteria. The primers include an upstream primer (see SEQ ID NO:1 for sequence) and a downstream primer (see SEQ ID NO:2 for sequence). The primers and kit can quickly and accurately identify tomato gray leaf spot pathogenic bacteria, and are of great significance for early detection and disease control of tomato gray leaf spots.

The invention discloses a method for rapidly detecting the polymorphism of a saccharum sucrose phosphate synthase B (SPSB) gene, belonging to the field of molecular breeding of plants. The method comprises the steps of designing a pair of primers to carry out PCR amplification by taking the genome DNA of saccharum robustum B.51NG3, saccharum barberi J.Nagori, saccharum species ROC22, saccharum.officinarum L.Badila, saccharum sinense R. lustrous bamboocane or saccharum spontaneum L.yinge No.1 as a template; carrying out agarose gel detection to name the SPSB genotype of a fragment with the size of 287bp as type C1 and name the SPSB genotype of a fragment with the size of 331bp as type C2; and identifying the polymorphism of the saccharum SPSB gene according to different sizes of the fragments. The invention provides a DNA level identifying method for saccharum sugar trait marker-assisted selection, and the DNA level identifying method is beneficial to tracking and utilization of different haplotypes of SPSB genes in filial generations of saccharum and has important significance for rapidly establishing saccharum germplasm with excellent heredity.

The invention relates to a primer probe composition and a kit for synchronously detecting 29 pathogens related to digestive tract infection of children. According to the primer probe composition and the kit, a primer group, a probe group and the kit can realize simultaneous detection of a plurality of common bacteria, viruses, fungi and parasitic pathogens causing digestive system infection by onereaction, the method is the most comprehensive children digestive tract infection detection method at present, the efficiency is high, the specificity is strong, and an auxiliary diagnosis basis is provided for clinical examination. By means of the technical scheme, the technical problem that in the prior art, diagnosis and research on pathogens related to digestive tract infection of children are still insufficient can be solved.

The invention relates to a primer and probe combination and kit for detecting 9 children digestive tract pathogens. A primer group, a probe group and the kit can realize simultaneous detection of a plurality of common viruses causing digestive system infection by one reaction, and a method is the most comprehensive detection method for children digestive tract virus infection at present, and is high in efficiency and strong in specificity, and an auxiliary diagnosis basis is provided for clinical examination. By means of the technical scheme, the technical problem that in the prior art, diagnosis and research on virus pathogens related to children digestive tract infection are still insufficient can be solved.

The invention provides a multiple RT-PCR detection method, which can simultaneously detect whether leaves, twigs, flowers or fruits of an apple tree bring three latent viruses namely apple chlorotic leaf spot virus, apple stem grooving virus, and apple stem pitting virus, and three apple scar skin viroids namely apple scar skin viroid, apple dimple fruit viroid, and apple fruit crinkle viroid or not. The detection method has the advantages of clear detection result, convenient operation, rapid reaction, and high sensitivity, and overcomes the shortages of time consuming, tedious steps, difficulty in distinguishing the stripes, and low sensitivity in the prior art. The invention provides a rapid detection method, which is easy for promotion and application, for detecting apple viruses.

The invention discloses a serum / plasma exosome miRNA marker related to atrial septal congenital heart disease. The application of the product in heart disease can alleviate the technical problem of insufficient research on specific miRNA related to atrial septal congenital heart disease existing in the prior art.

The invention discloses a molecular marker for identifying trichotylenchus changlingensis, belongs to the field of plant nematode identification, and the molecular marker is a DNA segment of 520bp. The invention also discloses a PCR (polymerase chain reaction) primer of the molecular marker which is formed by the nucleotide sequences shown as the SEQ ID No. 2 and SEQ ID No. 3. The invention further discloses a primer and a molecular marker of double PCR for identifying the trichotylenchus changlingensis, and a corresponding double PCR identification method. The primer designed by aiming at the trichotylenchus changlingensis, which is provided by the invention, is high in specificity; the molecular marker obtained by amplifying the primer is high in accuracy and reliability when being used for indentifying the trichotylenchus changlingensis; furthermore, the method provided by the invention has the advantages that the detection sensitivity is high, the method is simple, the detection speed is high and the efficiency is high; in addition, the requirement to operators is low.

The present invention relates to a cercospora edulis specific molecular detection primer and application thereof, the cercospora edulis specific molecular detection primer includes an upstream primerand a downstream primer as shown in sequence tables SEQ ID No.1-2. The cercospora edulis specific molecular detection primer pair can amplify specifically in cercospora edulis to obtain a target product of 769 bp. The application comprises application of the cercospora edulis specific molecular detection primer in detection of soybean Cercospora sojina Hara and preparation of a detection kit for detecting the soybean Cercospora sojina Hara. The primer has high specificity and can be used for distinguishing soybean sclerotinia blight, Soybean Phytophthora Root, Soybean purple blotch, soybean gibberellic disease, soybean wilt and other soybean common disease pathogenic bacteria, and has high sensitivity, the detection sensitivity of cercospora edulis genomic DNA is up to 1ng / mu L; the usemethod is simple and quick, and cloning, sequencing, enzyme digestion and other operation are not needed for PCR products.

The invention discloses a method for quickly detecting salmonella through loop-mediated isothermal amplification. The method comprises the following steps: extracting the genome DNA (deoxyribonucleicacid) of bacteria to be detected: taking the extracted genome DNA as a template to carry out LAMP (Loop-mediated isothermal amplification), wherein the final concentration of magnesium sulfate in an amplification reaction system is 6.5-8mM / L, the final concentration of dNTPs (High quality deoxyribonucleotide triphosphates) is 4.2-5mM / L, the final concentration of F3 and B3 primers is 0.13-0.26mu m / L, the final concentration of FIP (Forward Inner Primer) and BIP (Backward Inner Primer) primers is 1.2-2 mu M / L, the final concentration of Bst DNA polymerase is 0.3-1.8U / mu L, the final concentration of Betaine is 1-2M / L, and amplification temperature is 60-62 DEG C for 1-1.5h and 85-87 DEG C for 1-3min. The method has the advantages of high primer specificity, high sensitivity and lower cost,whether a detected sample contains salmonella or not can be judged according to whether amplification is carried out or not, and detection can be quickly finished.

An objective of the present invention is to provide a molecular marker primer for identifying Trachitotus ovatus and Trachinotus blochii and application thereof, which can help solve the problems of mixed marine species and the like, and provide technical support for pompano breeding, pompano resource conservation, rational use and biodiversity research. The molecular marker primer for identifying Trachitotus ovatus and Trachinotus blochii includes a primer CSF and a primer CSR, and specific sequences are as follows: CSF: GTCATACGCTCCCGAAGAT; and CSR: TAGGGCTAAGCATAGTGGG. The molecular marker primer is used for identifying Trachitotus ovatus and Trachinotus blochii; and a kit including the molecular marker primer and a PCR reaction system reagent is applied to the identification of Trachitotus ovatus and Trachinotus blochii. The application also includes that molecular markers are used for identifying Trachitotus ovatus and Trachinotus blochii, and the application also includes identifying fries thereof According to results, the two fishes have a difference of 2 bases, which is shown as 1 transformation and 1 transversion. The results are stable and the reproducibility is high.