71 results about "Group specific primers" patented technology

The invention discloses a human leukocyte antigen HLA-A and HLA-B gene full-length sequencing method and an HLA gene sequencing and typing method. The HLA-A and HLA-B gene full-length sequencing method comprises the following steps of: a, performing PCR amplification on about 4kb full-length sequences of HLA-A and HLA-B genes by using a pair of primers respectively; and b, cloning the amplification products to a pGEM-Tea sy vector, sequencing the full-length sequences by using ten walking sequencing primers in positive and negative directions respectively, and totally obtaining 38 allele 4.3kb full-length sequences of the HLA-A and 30 allele 3.7kb full-length sequences of the HLA-B. The HLA-A and HLA-B sequencing and typing method comprises the following steps of: performing PCR amplification on typing target areas of the HLA-A and HLA-B by using two pairs of primers respectively; and performing two directional sequencing on products by using fourteen sequencing primers respectively, wherein the HLA-DRB1 and HLA-DQB1 sequencing and typing method comprises the following steps of: amplifying sequences of second and third exons of DRB1 and DQB1 by adopting group specificity primers respectively; performing two directional sequencing on the second and third exons of the DRB1 by adopting eight group specificity primers and three sequencing primers; and performing two directional sequencing on the second and third exons of the DQB1 by adopting four sequencing primers respectively.

The invention relates to a method for identifying leech by one group of specific primers and belongs to the technical field of traditional Chinese medicine identification. The method comprises the following steps: firstly, preparing a DNA test sample solution; secondly, taking the DNA test sample solution as a template, selecting specific nucleotide sequence obtained through design and screening as primers to perform PCR amplification; thirdly, performing agarose gel electrophoresis analysis and gel imaging system detection on products; and finally, judging the true and false and the adulteration condition of the leech according to the detection result. The specific primer amplification method is simple in operation, sequencing is not needed, the result is intuitive, and the adulteration product can be detected and identified; meanwhile, the method is high in anti-interference ability and high in repeatability, can be used for accurately identifying leech foundation and medicinal materials, provides an efficient and rapid method for detecting the leech products, and has a relatively good application value.

The invention discloses a duplex fluorescent RT-LAMP detection group and kit for visually identifying foot-and-mouth disease virus and Bluetongue virus and an application of the detection group and the kit. The duplex fluorescent RT-LAMP detection group comprises two groups of specific primers and probes, the first group of specific primers and probes comprise FMDV-F3, FMDV-B3, FMDV-FIB(F1c-F2), FMDV-BIP(B1c-B2) and FMDV-probes, the second group of specific primers and probes comprise BTV-F3, BTV-B3, BTV-FIB(F1c-F2), BTV-BIP(B1c-B2) and BTV-probes, and the two groups of specific primers and probes are as shown in a sequence table SEQ ID No.1 to SEQ ID No.10. The duplex fluorescent RT-LAMP detection group can identify and diagnose the foot-and-mouth disease virus and the Bluetongue virus inthe same reaction tube and has the advantages of good specificity, high sensibility, less pollution, convenience, rapidness and the like, a detection result can be directly observed by naked eyes andis judged according to colors of reaction products, and the detection group can be used for basic-level quarantine with poor conditions.

The invention relates to a rapid screening and detection technology for glyphosate-tolerant transgenic soybean and processed products, in particular to primer groups for detecting the commonly used inserted gene EPSPS of glyphosate-tolerant transgenic plants. Aiming at the EPSPS gene and according to the conserved sequence of the EPSPS gene, the invention designs two groups of specific primers. By adopting the two groups of specific primers and the loop-mediated isothermal nucleic acid amplification technology, the EPSPS gene can be rapidly, sensitively and specifically detected, so the products containing glyphosate-tolerant transgenic plant ingredients are screened. The primer can be provided with other reagents in the form of reagent kits and is used for nucleic acid amplification reaction. The method has the advantages of simple and convenient operation and good repetitiveness.

The invention discloses a probe and primer composition for rapidly detecting seven coronaviruses and other respiratory tract pathogens. A group of specific primers and the Taqman or MGB probe are designed mainly according to the specificity of N genes of OC43-CoV, NL63-CoV, 229E-CoV, HKU1-CoV, SARS-CoV, FluB and RSV, ORF1a, ORF1b and N genes of MERS-CoV, ORF1ab and N genes of 2019-nCoV, an M geneof FluA and an HEXON gene of HadV, and by utilizing a melting curve technology, CoV-OC43, HCoV-NL63, HCoV-229E, HCoV-HKU1, SARS-CoV, MERS-CoV, 2019-nCoV, FluA, FluB, HAdV and RSV are rapidly identified according to the change of the melting temperature. By means of the design, multiple pathogens can be detected in one fluorescent channel at the same time, and a relatively high sensitivity can be met.

The invention discloses a quick molecular detection method for simultaneously detecting three kinds of fusarium toxins and application thereof, is special for detecting three kinds of common fusarium toxins, and belongs to the technical field of plant quarantine, in particular to the technical field of molecular detection of the fusarium toxins. A plurality of composite primers are designed by the invention according to the closely related Trill gene which is synthesized by the toxins in the fusarium genome; the specific segments of the fusarium toxins nivalenol (NIV), 15 acetyldeoxynivalenol (ADON) and 3ADON are obtained from the fusarium collected in China and foreign countries through detection and separation; and the overall lengths of the specific segments of the NIV, 15ADON and 3ADON are 643bp, 424bp, and 342bp respectively. Only one group of specificity primers are used for performing polymerase chain reaction (PCR) amplification reaction, the PCR product is detected by 1.5 percent agarose gel electrophoresis, and the types of the three kinds of B type toxins produced by fusarium graminearum schw can be directly detected according to the length of the product segments; and the method can be applied to the detection of the deoxynivalenol (DON) and NIV toxin pollution in grain, feed and food safety.

The invention belongs to the technical field of bovine virus detection and particularly relates to a foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP (loop-mediated isothermal amplification) detection primer group, a kit and application thereof. The foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP detection primer group comprises two groups of specific primers, wherein one group is FMDV-F3, FMDV-B3, FMDV-FIP (F1c-F2) and FMDV-BIP (B1c-B2), and the other group is VSV-F3, VSV-B3, VSV-FIP (F1c-F2) and VSV-BIP (B1c-B2), and sequences of the two groups are shown as SEQ ID NO.1 and SEQ ID NO.8 respectively. An established foot and mouth disease virus and vesicular stomatitis virus identifying duplex fluorescence RT-LAMP method has advantages of simplicity, convenience, quickness, specificity, sensitivity and the like and can be used for FMDV (foot and mouth disease virus) and VSV (vesicular stomatitis virus) clinical detection and epidemiological investigation. The FMDV and VSV duplex fluorescence RT-LAMP method is a simple, quick and low-cost diagnosis method and suitable for large-scale epidemiological investigation.

The invention provides a group-specific primer PCR-SBT reagent based on HLA-DQB1 genetic typing. The group-specific primer PCR-SBT reagent is composed of six pairs of group-specific primers for amplification and four oligonucleotides sequencing primers for sequencing analysis. The invention furthermore provides a group-specific primer PCR-SBT method based on HLA-DQB1 genetic typing adopting the reagent. The reagent and method can be used as an independent widely-applied identification method. The accurate typing identification problem of HLA-DQB1 sites can be successfully solved. Accuracy of HLA matching of hematopoietic stem cell transplantation donors and recipients can be improved. Therefore, more proper transplantation donors are selected, and rejection reactions in the transplantation process are reduced. Great significance in furthermore improving the success rate and survival rate of organ transplantation is achieved.

The invention discloses a method for detecting hantaan virus strain genome. The method has the following steps: (1) two pairs of hantaan virus general primers are designed; (2) the hantaan virus detection method comprises the following steps that: first, viral specific RNA extraction kit is utilized to extract the total RNA in a serum sample; second, a nucleic acid analyzer is adopted to detect the RNA content, and the RNA is taken to perform the RT-PCR reaction; third, RT-PCR:RT uses HV group-specific primers such as P0, PCR uses HV general primers such as P1P2 and P3P4; fourth, viral nucleic acid amplified results are observed through agarose gel electrophoresis, and a specific nucleic acid band appears. The method is intuitionistic and predominant, the sensibility is good, and the effects are good; the serum detection rate in recent ten years is 79.2 percent, and the serum detection rate in twenty years ago can still reach to 70.5 percent; the method is applied to the laboratory detection of hantaan virus infection and the molecular epidemiological survey.

The invention relates to a fluorescent quantitative detection kit for simultaneously detecting human influenza A virus, human influenza B virus and novel coronavirus, and belongs to the technical field of nucleic acid detection. The detection kit specifically comprises four groups of specific primer pairs and probes, a negative quality control material, a positive quality control material and a fluorescent quantitative PCR reaction system, wherein the four groups of specific primer pairs and probes are respectively a pair of specific primers and a probe for detecting influenza A virus and influenza B virus, and two pairs of specific primers and two probes for detecting novel coronavirus; wherein the positive quality control material is an artificially synthesized target sequence, and the negative quality control material is deionized water; wherein the fluorescent quantitative PCR reaction system is composed of components for PCR reaction and reaction conditions. According to the kit disclosed by the invention, the specific primer pair and the probe are designed at a conservative site of a virus sequence; the kit can realize simultaneous detection of influenza A virus, influenza Bvirus and novel coronavirus in a single tube, has strong detection specificity and high sensitivity, and can accurately and rapidly distinguish influenza from new coronapneumonia patients from people,so as to realize early discovery, early isolation and early treatment. The kit has important application value in the fields of disease monitoring, clinical diagnosis and the like.

The invention belongs to a qualitative detection technology of plant derived components in the foods and the beverages, in particular to a primer / a probe group for detecting a carrot component in the foods and the beverages. In the invention, a group of specific primer and probe are designed aiming at antifreeze protein of the carrot and according to the conserved sequence of a gene thereof. The carrot component in the foods and the beverages can be rapidly, sensitively and specifically detected by using the primer and the probe and adopting a real-time fluorescence PCR technology. The primer and the probe can be provided with other reagents together in the form of a kit and are used for amplification reaction of nucleic acid. The method has simple operation and good repeatability.

The invention belongs to technology of qualitative detection of animal-derived components in food and feed, and particularly relates to a primer / probe group for detecting fox component in food and feed. The invention selects mitochondrion cytochrome C redox enzyme I subunit gene sequence of fox, and designs a group of specific primers and probes. After using the primers and probes, the real-time fluorescent PCR (polymerase chain reaction) technology can be adopted to quickly, sensitively and specifically detect the fox component in food and feed. The primers and probes can also be provided together with other reagents in the form of a kit, and is used for nucleic acid amplification reaction. The method is simple to operate and has favorable repetitiveness.

The invention belongs to the field of biotechnologies, and particularly relates to a multi-PCR-ELISA detection kit for human seasonal influenza viruses and an application method thereof. The detection kit disclosed by the invention is composed of a RT-PCR reaction system, an ELISA detection system, four target-gene (an influenza-A-virus M gene, a H1 subtype virus HA gene, a H3 subtype virus HA gene, and a B type virus NS gene) positive plasmids (Pa-m, Ph1-ha, Ph3-ha and Pb-ns), a negative quality control specimen and four target-gene specific primer probes. Specific amplification is performed by using specific primers labeled by using four sets of biotins through RT-PCR, an amplified product after being denatured is hybridized with a specific probe labeled by using digoxin, a hybridized product is enveloped with a streptavidin-enveloped 96-hole micro-plate, and an anti-digoxin antibody labeled by using horse radish peroxidase is added for carrying out detection through an ELISA method, so that a rapid, sensitive and specific typing detection kit for seasonal influenza viruses such as H1 and H3 subtypes and hepatitis B viruses is established. The invention relates to the application of four sets of specific primer probes in the clinical differential diagnosis of influenza virus infection and the typing authentication of influenza virus isolates.

The invention discloses a primer and a probe for detecting racoon dog components in foods and feeds and belongs to a qualitative detection technology of animal origin components in the foods and the feeds. According to the primer and the probe, a set of a specific primer and a probe are designed by selecting mitochondria cell pigment C oxidordeuctase I subunit gene sequence; the primer and the probe are used and a real-time fluorescence PCR (Polymerase Chain Reaction) technology is adopted so as to rapidly, sensitively and specifically detect the racoon dog components in the foods and the feeds. The primer and the probe can be provided in a form of a kit with other reagents and are used for a nucleic acid amplification reaction. The primer and the probe are simple and convenient to operate and have good repeatability.

The invention provides a heterodera filipjevi Taqman MGB probe real-time quantitative PCR detection method and application thereof, belongs to the technical field of plant nematode molecular detection, and discloses a specific fluorescence probe HF-probe and a group of specific primers qHF-R and qHF-F, which are designed on the basis of a heterodera filipjevi RAPD specific sequence. The probe and the primers can be amplified in heterodera filipjevi to obtain specific fluorescence curves, so that the quick molecular detection of heterodera filipjevi is achieved. The detection method has high sensitivity and strong specificity, is fast and accurate, and has high practical applied values in the aspects of early diagnosis and field monitoring and early warning of heterodera filipjevi.

The invention relates to qualitative detection methods of ingredients of animal origin in food, forage, fur, and products of food, forage and fur, and more specifically relates to primers and probes used for detecting ingredients of racoon dog origin, and a detection method of the ingredients of racoon dog origin. According to the detection method, gene sequences of mitochondrion cytochrome oxidase b are selected, a set of specific primers and probes is designed, and the specific primers and probes are used for real-time fluorescent PCR, so that simple, convenient and fast detection of the ingredients of racoon dog origin in food, forage, fur, and the products of food, forage and fur is realized.

The invention pertains to a test technology of transgenic ingredients in food and feed, and specifically relates to a primer pair used for testing a common promoter used for transgenic plants in food and feed, namely, cauliflower mosaic virus (CaMV) 35S promoter. The invention designs a pair of specific primers, aiming at the cauliflower mosaic virus (CaMV) 35S promoter and according to the genetic conserved sequence thereof. The primer pair is used and a loop-mediated isothermal amplification technology is adopted for rapidly, sensitively and specifically testing the CaMV 35S promoter, thus screening whether the products contain transgenic plant ingredients. The primer can also be provided in kit and together with other reagents and used for nucleic acid amplification. The method has simple and convenient operation and good repeatability.

The invention relates to a quick qualitative detection technology on Zygosaccharomyces bailii in grape wine and fruit juice and in particular relates to a primer and a probe which are used for Zygosaccharomyces bailii detection and a detection method thereof. According to the invention, a 26S ribosome RNA gene sequence of Zygosaccharomyces bailii is selected, a specific primer and probe set is designed, and simple and quick detection on Zygosaccharomyces bailii in the grape wine and fruit juice can be realized by adopting the primer and the probe and a real-time fluorescent PCR technology.

The invention discloses a primer set for quickly, qualitatively and quantitatively detecting six common intestinal microorganisms, which consists of six groups of specific primer pairs respectively used for detecting pseudomonas, lactobacillus, escherichia coli, enterococcus, streptococcus and bifidobacterium. PCR detection by using the primer set combined with agarose gel electrophoresis can carry out quantitative analysis on a sample; the primer set is used for fluorescence quantitative PCR detection to carry out quantitative detection on the sample. The primer set can be carried out under the same PCR reaction program, can carry out qualitative and quantitative detection on six common microorganisms in the intestinal tract within four hours, and has wide application prospect in the fields of medical detection, food sanitation and the like.

The invention belongs to the field of qualitative detection technology for plant derived components in food and beverage, and specifically provides a primer / probe group for detecting garlic component in food and beverage. According to the invention, a group of specific primers and a probe are designed based on a conservative sequence of an allinase gene in garlic. The primer group and the probecan be employed in real-time fluorescent PCR technology to detect garlic component in food and beverage rapidly, sensitively and specifically. The primer group and the probe can be provided together in the form of a kit or other reagents, and applied to nucleic acid amplification reaction. The method has the advantages of simple operation and good repeatability.

The invention belongs to a qualitative detection technology of animal-derived ingredients in feather down products, and specifically relates to a primer / probe group for detecting an alpaca component in a fiber product. A group of specific primer and probe is designed by selecting a mitochondrial COXI gene sequence of alpaca. By using the primer and the probe and adopting a real-time fluorescence PCR (polymerase chain reaction) technology, the alpaca component in the fiber product can be fast, sensitively and specifically detected. The primer and the probe can be provided together with other reagents in the form of a kit and be used for nucleic acid amplification reaction. The method is simple and convenient to operate and good in repeatability.

The invention discloses a non-obstructive azoospermia auxiliary diagnosis gene detection kit, relates to the field of azoospermia diagnosis, and solves the problem of difficult clinical diagnosis of male infertility. The non-obstructive azoospermia auxiliary diagnosis gene detection kit comprises a specific PCR primer, a PCR buffer solution, dNTPs and DNA polymerase, wherein the specific PCR primer is a group of specific primers for detecting the 69826819 mutation site in the non-obstructive azoospermia TEX11 gene. A specific primer SEQ ID NO: 1 and a specific primer SEQ ID NO: 2 are designed,the specific primers have higher specificity, the non-obstructive azoospermia auxiliary diagnosis gene detection kit consisting of the specific primers is adopted to detect the 69826819 mutation sitein the TEX11 gene of a male patient, and the detection rate of the gene mutation is 100%.

The invention provides a triple fluorescent quantitative PCR kit for simultaneously detecting a bovine rotavirus, a bovine coronavirus and a bovine viral diarrhea virus and an application, and belongs to the technical field of virus detection. The triple fluorescent quantitative PCR kit comprises three groups of specific primers and three corresponding probes aiming at the bovine rotavirus, the bovine coronavirus and the bovine viral diarrhea virus. The primer and the probe for detecting the three viruses have the advantages of strong specificity and high reaction efficiency. The triple real-time fluorescent quantitative PCR kit has the characteristics of high amplification efficiency, strong specificity, stable detection effects and good repeatability, and is suitable for dairy cow diarrhea pathogen detection, regular monitoring, dairy cow diarrhea epidemiological investigation and the like. The kit disclosed by the invention can be used for simultaneously detecting three dairy cow diarrhea pathogens in a real-time fluorescent quantitative PCR reaction system, so that the detection time is saved, the detection cost is reduced, a plurality of samples can be simultaneously detected, and the detection efficiency is high.

The invention relates to a specific primer composition for lumpy skin disease viruses, and also relates to a kit and a method for detecting the lumpy skin disease viruses. At least two groups of specific primer pairs are designed for conserved sequences of lumpy skin disease viruses, and a detection kit and a detection method are further established. The kit and the method provided by the invention have the advantages of high sensitivity, shorter amplification time and higher efficiency, can identify whether the lumpy skin disease virus contained in the sample comes from vaccine immunity or natural infection, and are a powerful supplement to the existing lumpy skin disease virus detection method.

The invention belongs to a qualitative detection technology of animal-derived ingredients in feather down products and specifically relates to a primer / probe group for detecting a goose down component in the feather down product. A group of specific primer and probe is designed by selecting a mitochondrial cytochrome B (CYTB) gene sequence of goose down. By using the primer and the probe and adopting a real-time fluorescence PCR (polymerase chain reaction) technology, the goose down component in the feather down product can be fast, sensitively and specifically detected. The primer and the probe can be provided together with other reagents in the form of a kit and be used for nucleic acid amplification reaction. The method is simple and convenient to operate and good in repeatability.

The invention relates to molecular detection of Arceuthobium sichuanense and belongs to the field of biotechnology. Based on sequence differences between clpP genes of Arceuthobium sichuanense and its host dragon spruce and by comparing sequence differences between Arceuthobium sichuanense and its host dragon spruce, a group of specific primers (4 primers) are designed. Meanwhile, the invention provides a group of LAMP primer sequences for detecting Arceuthobium sichuanense and also further provides a detection method for detecting Arceuthobium sichuanense. Primers specificity is good. The detection method is fast and simple, has high accuracy and provides an effective technical support for early diagnosis and disease control of Arceuthobium sichuanense.

The invention discloses a primer composition, a kit and a method for detecting mycoplasma bovis and infectious bovine rhinotracheitis viruses. The primer composition designed and screened in the invention comprises two groups of specific primers and probes. Under the condition of an optimized reaction system and under optimized reaction conditions, mycoplasma bovis and infectious bovine rhinotracheitis viruses can be simultaneously identified and detected in the same reaction tube; and the primer composition, the kit and the method have the characteristics of good specificity, high sensitivity, interference resistance, no pollution, convenience and quickness in operation, low cost and the like.

The invention provides a group of specific primers with high sensitive for detecting botryosphaeria dothidea of kiwi fruit. The group of specific primers comprise a forward primer and a reverse primer, wherein the forward primer is 5'-TCATTCGTGATCTCAAGCCAGA-3' and is as shown in SEQ ID No.1, the reverse primer is 5'-TGCTGGTGAAGATAGACATCTGC-3' and is as shown in SEQ ID No.2. The invention further provides a detection method and application based on the specific primer.