310 results about "Fluorescent pcr" patented technology

The invention relates to the field of biological detection, in particular to a primer group and a kit for fusing round cell sarcoma by using EWSR1 non-ETS and a use method. The EWSR1 non-ETS fused round cell sarcoma detection primer group comprises nine primer sequences, namely a primer group A and a primer group B. The primer group A and the primer group B are used for detecting EWSR1 non-ETS fused round cell sarcoma. According to the invention, the variation types of the EWSR1 non-ETS fusion sarcoma are divided into five variation types, the five fusion variation types are divided into two groups, two groups of upstream and downstream amplification primer groups are designed in a targeted manner, and each fusion variation type can be amplified to obtain a target product peak with the size of 100-250 bp, so that the target product peak can be amplified to obtain a target product peak with the size of 100-250 bp. The primer provided by the invention can be used for detecting five variation types fused by three types of genes at one time. Through combined application of the primer group, multiple fluorescent PCR and high-resolution capillary electrophoresis, the lowest detection limit is 10 copies, and the sensitive and efficient detection method for the EWSR1 non-ETS fused round cell sarcoma is provided and has the characteristics of simplicity, convenience, accurate result and economic benefit.

The invention discloses a nucleic acid product, a kit and a method for detecting various pathogens. The nucleic acid product comprises a primer pair and a probe, wherein the nucleotide sequences of the primer pair and the probe are respectively shown as SEQ ID NO: 1-24. Specific primer probes are respectively designed for conserved regions of eight respiratory tract pathogenic bacteria, synchronous detection of multiple pathogens is realized through a one-tube eight-color fluorescent PCR, freeze-drying and ultrasonic direct diffusion method technology, experimental operation steps are remarkably reduced, the detection time is shortened, the result interpretation complexity is reduced, and in addition, the detection efficiency is greatly improved. The anti-interference capability is strong, and the sensitivity is high.

The invention discloses a respiratory infectious disease multi-pathogen nucleic acid detection kit based on a fluorescent PCR detection technology and application of the respiratory infectious disease multi-pathogen nucleic acid detection kit. The invention provides two detection kits aiming at 20 and 6 respiratory tract detection, the two kits can specifically detect respiratory tract pathogens, the detection sensitivity is high, the whole detection system is short in time consumption and simple and convenient in use process, and detection results of 20 pathogenic microorganisms can be reported within 40 min at the shortest; the whole set of detection kit and the detection method are high in sensitivity and good in specificity; the normal-temperature storage and transportation of the reagent can be realized.

The embodiment of the invention discloses a probe primer combination for identifying TaqMan real-time fluorescent quantitative PCR of ampullaria gigas, a kit and application. Comprising a probe, an upstream primer and a downstream primer, the nucleotide sequence of the probe is 5 '-FAM-CCTTCTCTATTACTACTATTA-MGB-3', the nucleotide sequence of the upstream primer is 5 '-CTTTCCGCGTCTAATAACATG-3', and the nucleotide sequence of the downstream primer is 5 '-CATCAGTTCCAGCACCACTC-3'. The TaqMan probe real-time fluorescent PCR probe and primer combination established by the invention has excellent specificity and sensitivity in the aspect of identifying the species of the ampullaria gigas, the detection accuracy is greatly improved, the detection time is shortened, the effects of stability, reliability, short period and low cost are achieved, and the primer combination has good application prospects in rapid and accurate identification of the ampullaria gigas. The method is of great significance in promoting related prevention and research work.

The invention designs a carbapenem drug-resistant gene nucleic acid fluorescent PCR method detection kit and application thereof, the kit uses Taqman fluorescent probe multiple amplification technology to detect carbapenem drug-resistant genes, the kit comprises a nucleic acid reaction liquid Mix1 and a nucleic acid reaction liquid Mix2, the nucleic acid reaction liquid Mix1 is used for detecting drug-resistant genes IMP, NDM and OXA48, the nucleic acid reaction liquid Mix2 is used for detecting drug-resistant genes IMP, NDM and OXA48, and the nucleic acid reaction liquid Mix2 is used for detecting drug-resistant genes IMP, NDM and OXA48. The nucleic acid reaction liquid Mix2 is used for detecting drug resistance genes VIM, KPC and OXA23. The method is high in detection specificity, and the detection result is visual and easy to interpret. Clinical doctors are assisted to judge respiratory tract pathogen infection more comprehensively and more accurately. The kit covers six carbapenem drug resistance genes, is more comprehensive and accurate in detection, and is high in specificity and sensitivity. Meanwhile, multiple sample types can be detected, a sputum sample, an excrement sample and a rectum swab sample are detected through paramagnetic particle extraction, and pure bacterial colonies are directly detected without extraction.

The invention relates to the technical field of molecular biological detection, and particularly discloses a quadruple fluorescent PCR primer probe combination, a reaction system and a detection method for identifying a mycoplasma gallisepticum wild strain and a vaccine strain. The quadruple fluorescent PCR primer probe combination for identifying the mycoplasma gallisepticum wild strain and the vaccine strain comprises a universal primer probe group for detecting mycoplasma gallisepticum and a specific primer probe group specially aiming at three vaccine strains F-36, TS-11 and 6 / 85 respectively. According to the quadruple fluorescent PCR primer probe combination, the reaction system and the method for identifying the mycoplasma gallisepticum wild strain and the vaccine strain, the purpose of simultaneously detecting and distinguishing DNA of the mycoplasma gallisepticum wild strain and DNA of three different vaccine strains (F-36, TS-11 and 6 / 85) of the mycoplasma gallisepticum wild strain can be achieved, so that detection and differential diagnosis are more efficient, economical, simple and convenient; the kit has the advantages of high sensitivity, good specificity, rapidness, high flux, simplicity and convenience in operation and the like.

The invention discloses a composition, a kit and a method for detecting a mitochondrial gene of sea eel. A primer probe composition targeting the D-LOOP gene of the sea eel mitochondria is designed, and the nucleotide sequences of the primer probe composition correspond to SEQ ID No.1-3. On the basis, the invention further constructs and optimizes a specific real-time fluorescent PCR detection method for the D-LOOP gene of the sea eel mitochondria. According to the method, a part of sequence of the D-LOOP gene of the mitochondria of the sea eel is taken as a target region, a primer and a TaqMan-MGB probe are designed in a targeted manner, and finally a set of rapid, sensitive and high-specificity detection system is established. The detection system can be effectively applied to adulteration and germplasm identification work of the sea eel, and has a very wide industrial application prospect.

The invention relates to the technical field of bacterial detection, and aims to provide a method for detecting staphylococcus aureus through real-time fluorescent PCR and application. The method comprises the following steps: (1) collecting a sample; (2) extracting a genome of a sample to be detected: extracting the collected sample, and taking a certain amount of the genome of the final product sample as a subsequent template; (3) real-time fluorescent PCR detection: taking the genome in the step (2) as a template, preparing a reaction system by using a primer and a probe, setting an amplification reaction program, and carrying out real-time fluorescent PCR detection; and (4) result interpretation: judging whether the to-be-detected sample is infected with the staphylococcus aureus or not according to the real-time fluorescence PCR detection result. The detection method has the advantages of high sensitivity, strong specificity and accurate quantification, and is suitable for detecting staphylococcus aureus in a large number of samples.

The invention relates to a multiplex PCR (Polymerase Chain Reaction) primer probe combination, a kit and a detection method for human red blood cell RHCE genetic typing detection, and belongs to the field of biomedicine clinical molecular detection. The invention provides a multiplex PCR (Polymerase Chain Reaction) primer probe combination for human red blood cell RHCE genetic typing detection, and further provides a kit containing the multiplex PCR primer probe combination. The method has the technical advantages that the RHCE gene variation site specific primer and probe are designed through an improved ARMS technology and a homologous sequence specific base method, so that higher specificity and accuracy are realized; only four-hole reaction is needed, detection of RHC, RHc, RHE, RHe, CW- / CW +, CX- / CX +, V-VS- / V + VS + and V + / V-can be completed at a time, and higher flux and efficiency are achieved; according to the method, multiple fluorescent PCR is combined with a Taqman probe technology, qualitative typing detection is carried out on the human red blood cell RHCE gene, and compared with a traditional method, higher sensitivity, higher experiment speed, higher accuracy and a wider application range are shown.

The invention discloses a kit for detecting HLA-B27 and a detection method. The kit for detecting HLA-B27 comprises a PCR (Polymerase Chain Reaction) solution, a primer probe mixed solution, a positive reference substance and a negative reference substance, the primer probe mixed solution comprises an upstream primer 1F, a downstream primer 1R and a probe 1P for detecting the HLA-B27, and an upstream primer 2F, a downstream primer 2R and a probe 2P for detecting a reference gene. A fluorescent PCR-melting curve method is adopted, sequence alignment is carried out according to an NCBI database, primers and probes which are high in specificity and capable of simultaneously detecting main subtypes (HLA-B27: 02, HLA-B27: 04, HLA-B27: 05, HLA-B27: 06, HLA-B27: 07 and HLA-B27: 24) of the HLA-B27 are selected, and compatibility detection of different subtypes of the HLA-B27 can be achieved through a pair of primers and a probe. The Tm value in the melting curve instead of the Ct value of the amplification curve is adopted for result judgment, qualitative accuracy is achieved, and the result is visual and easy to interpret.

The invention belongs to the field of molecular biological detection, and provides a joint detection composition which mainly utilizes a multiple fluorescent PCR analysis method to detect related pathogens, so that detection and distinguishing of at least four of clostridium praerzii, eubacterium rectum, streptococcus salivarius, clostridium tenella, prevotella and escherichia coli are simultaneously realized in a single-tube reaction system. Meanwhile, the detection sensitivity of the composition is higher and reaches 500 copies / mL, and the detection is more accurate.

The invention discloses a detection kit and method for Mycobacterium tuberculosis complex and rifampicin drug-resistant mutation, and belongs to the technical field of biologication.A Taqman probe and a molecular beacon are combined, a fluorescent signal is generated based on a Taq enzyme hydrolysis fluorescent probe, the fluorescent signal is accumulated, and the molecular beacon is used for detecting the drug-resistant mutation of the Mycobacterium tuberculosis complex and the rifampicin drug-resistant mutation of the Mycobacterium tuberculosis complex and the rifampicin drug-resistant mutation. According to two technical principles that a molecular beacon and a fluorescent product are hybridized to generate a fluorescent signal, fluorescent PCR amplification is adopted for detection of one target spot in a monochromatic fluorescent channel, and a melting curve mode of a probe and a target gene is adopted for detection of the other target spot. Therefore, two target spots can be detected and analyzed in a monochromatic fluorescence channel at the same time. Single-tube multiple detection is achieved, and mycobacterium tuberculosis complex group detection and mycobacterium tuberculosis rifampicin drug-resistant mutation site detection can be achieved at the same time.

The invention belongs to the field of molecular biological detection, and provides a joint detection composition which is mainly used for detecting related pathogens by using a multiple fluorescent PCR (Polymerase Chain Reaction) analysis method. Therefore, at least four of the klebsiella aerogenes, the stenotrophomonas maltophilia, the streptococcus angina, the acinetobacter baumannii, the klebsiella pneumoniae and the klebsiella oxytoca can be simultaneously detected and distinguished in a single-tube reaction system. Meanwhile, the detection sensitivity of the composition is higher and reaches 500 copies / mL, and the detection is more accurate.

The present application relates to the field of veterinary diagnostic technology, and specifically to a probe composition, a primer composition, and a triple fluorescence PCR kit and related methods for differential diagnosis of three genes of different strains of African swine fever composed of the above-mentioned probe composition and primer composition. The kit includes primer probes specific to the B646L gene, the CD2v gene, and the I177L gene. The present application designs fluorescent PCR primers and probes for the B646L gene, the CD2v gene, and the I177L gene, respectively, and establishes a triple fluorescence PCR detection method, which can quickly identify African swine fever virus strains and gene-deficient strains. At the same time, it has no cross-reaction with swine fever virus, porcine reproductive and respiratory syndrome virus, pseudorabies virus, porcine epidemic diarrhea virus, porcine circovirus, porcine parvovirus, healthy whole blood, oropharyngeal swabs, nasal swabs, anal swabs, environmental swabs, pig tissues, etc., and has good specificity.

The invention relates to an apolipoprotein E detection reagent and application thereof, and belongs to the technical field of biomedicine. The invention provides application of a plant derivative in preparation of an APOE epitope exposure agent. The plant derivative is selected from at least one of curcumin, emodin, EF24 and FLLL-32. The invention also provides an APOE epitope exposure agent which comprises a surfactant, salts, a reducing agent and plant derivatives. According to the present invention, the sample and the protein are pre-treated by using the epitope exposure agent containing the plant derivative to improve the sufficiency of the ApoE protein epitope exposure so as to achieve the accurate typing of the ApoE gene based on the immunological detection method and the accurate quantification of the proteins with different genotypes. Genetic typing of ApoE and quantitative detection of proteins with different genotypes are carried out based on an immunological detection method, and the genotype detection result is highly consistent with a fluorescent PCR (Polymerase Chain Reaction) result.

The invention belongs to the field of agriculture and plant quarantine, and particularly relates to a method for detecting phomopsis helianthi based on RPA / CRISPR-Cas12a, which comprises the following steps: S1, designing a detection composition: designing a composition for detecting phomopsis helianthi, the composition comprising a specific primer RPA-F / R nucleic acid sequence, a specific crRNA nucleic acid sequence and probes FL-ssDNA and LF-ssDNA; s2, extracting DNA: extracting the genome DNA of the diseased tissue of the sunflower to be detected; s3, DNA isothermal amplification: taking the genome DNA obtained in the step S1 as a template, adding a specific primer RPA-F1 / R1 into the template, and carrying out RPA amplification to obtain an amplification product; s4, CRISPR / Cas12a reaction: the amplification product obtained in the S2 and the crRNA are respectively mixed with FL-ssDNA and LF-ssDNA probes for a cleavage reaction, and a reaction product FL and a reaction product LF are obtained; and S5, visual detection: respectively placing the reaction product FL and the reaction product LF in a fluorescent PCR instrument and on a Cas12 special detection test strip for detection. The method is based on an RPA / CRISPR-Cas12a technology, by designing a specific primer and a crRNA nucleic acid sequence, the detection duration of the phomopsis helianthi is greatly shortened, the detection sensitivity is improved, and the method has very high use value in detection scenes such as customs and the like.

The invention provides a kit for detecting TP53 gene mutation. The kit for detecting TP53 gene mutation provided by the invention can accurately detect six TP53 gene mutation sites, namely R175H, R249S, R282W, R248Q, R273H and G245S, in genome DNA to be detected at the same time through a multiple fluorescent PCR reaction system and a multi-channel fluorescent signal, can accurately detect the six mutation sites, has ultrahigh amplification efficiency, detection efficiency, sensitivity and specificity, and can be widely applied to detection of TP53 gene mutation. The genotype of a sample with the genome DNA content of 1 ng can be accurately detected, the operation is simple and convenient, and the cost is low.

This application relates to the field of virus detection technology, specifically disclosing a real-time fluorescent PCR detection kit and method for H10N7 avian influenza virus. The kit includes sample lysis extract, sample washing solution I, sample washing solution II, and reaction premix for detecting H10N7 subtype avian influenza virus; the detection method is an integrated detection method. This application achieves simultaneous detection of H10 and N7 subtype AIVs by designing specific primers and probes. The kit prepared using these primers and probes not only improves detection efficiency but also significantly enhances sensitivity and specificity. The combined use of MAOPA technology and real-time quantitative PCR technology further improves the accuracy of the detection results, making this kit of this application of significant application value in avian influenza prevention and control.

The invention provides a reagent for detecting the proportion of vibrio in prawn seeds or prawn culture water in total bacteria. The reagent comprises a vibrio detection primer and probe, a bacteria detection primer and probe, Taq DNA polymerase, a dNTP mixture, UNG enzyme, a freeze-drying protective additive and a PCR buffer solution. According to the method, the specific DNA of the vibrio and the bacteria in the sample is quantitatively detected by utilizing the fluorescent PCR technology, the proportion of the vibrio in the total bacteria is obtained through the obtained quantitative detection data, the proportion of the vibrio in the total bacteria in shrimp seeds and shrimp culture water can be obtained in a short time, and guidance is provided for shrimp breeding and culture.

The utility model discloses a constant-temperature fluorescent PCR (polymerase chain reaction) nucleic acid detection kit for food-borne escherichia coli O157, and relates to the technical field of detection kits. The plurality of cutting parts are fixed at the lower end in the box body at equal intervals and are used for cutting the reagent tubes into a plurality of groups; the plurality of clamping devices are arranged above the cutting part in a sliding manner, and are used for clamping the reagent tubes; the lifting assembly is mounted in the box body in a manner of sliding up and down, and is positioned above the clamping device; the lifting assembly and the clamping device are arranged in a matched mode to clamp and fix a reagent tube when the cover plate assembly is buckled, meanwhile, the lifting assembly slides to the upper portion, the upper portion of the reagent tube is limited, the reagent tube is prevented from shaking in the transportation process, clamping and fixing of the reagent tube are relieved when the cover plate assembly is taken down, and therefore the reagent tube is prevented from being damaged. The reagent tube is convenient to take and place.

The utility model relates to the technical field of biological detection, in particular to a fluorescent PCR (Polymerase Chain Reaction) nucleic acid detection reagent strip for detecting Listeria monocytogenes, which is characterized in that after a sample to be detected is added into the reagent strip through a sample adding hole, the sample flows into a nucleic acid extraction area for nucleic acid extraction, and the extracted nucleic acid enters an amplification reaction area for amplification reaction; when a target nucleic acid sequence is amplified and then enters fluorescence detection to generate a fluorescence signal, a sensor in a fluorescence detection area captures the fluorescence signal in real time and transmits the signal to a control chip, the control chip processes and analyzes data, and a result is transmitted to external equipment (such as a computer, a mobile phone and the like) through a data transmission module. The rapid detection on the listeria monocytogenes is realized. The reagent strip disclosed by the utility model integrates the functions of nucleic acid extraction, amplification reaction and detection, is simple and convenient to operate, does not need complex sample pretreatment and professional experimental equipment, and is suitable for on-site rapid detection and basic application.

1. Name of the Design Product: Fully Automatic Nucleic Acid Purification and Real-Time Fluorescence PCR System. 2. Use of the Design Product: The design product is used for nucleic acid detection, mainly including an integrated process such as sample pretreatment, nucleic acid extraction, purification, real-time fluorescence PCR detection and report output. 3. Design Key Points of the Design Product: The combination of shape and pattern. 4. Picture or Photo Most Indicating the Design Key Points: Front View.

The present invention provides dual-target site reverse transcription fluorescent PCR primers, probes, and a kit for detecting the novel coronavirus SARS-CoV-2. By performing nucleic acid sequence alignment on the entire genome of the novel coronavirus SARS-CoV-2, three specific nucleic acid sequences W1, W2, and W3 in the viral N gene are found, and the sequences are shown as SEQ ID NOs. 1-3, respectively. Multiple pairs of primers and probes are designed for the three target sites, respectively. It is found that the dual-target site reverse transcription fluorescent PCR using a primer-probe combination designed for the W1 and W2 targets (sequences shown as SEQ ID NOs. 4-6 and SEQ ID NOs. 7-9, respectively) can specifically and sensitively detect the novel coronavirus SARS-CoV-2, with a detection sensitivity of less than 10 copies. There is no non-specific amplification of 22 common clinical respiratory pathogens, which can effectively avoid false negative results caused by single target site mutations and has good specimen detection capabilities.

The present application relates to the field of molecular biology, in particular to a primer probe composition, kit and detection method for 23 human papilloma virus genotyping detection, wherein the hydrolysis probe and molecular beacon probe are respectively designed in two different regions of HPV gene sequence E6 and L1, and the fluorescence collection point in the amplification stage is set in the high-temperature denaturation stage, so that the detection in the amplification stage and the melting curve stage is independent and does not interfere with each other, thereby realizing the combined use of real-time fluorescent PCR amplification detection technology and melting curve technology; the technology improves the single-well detection target flux of fluorescent PCR analysis technology, reduces the detection cost, and provides a more optimal detection method for HPV multiplex fluorescent PCR genotyping detection.

The invention discloses a spirochete-like multiplex nucleic acid detection kit and a matched artificial chromosome engineering bacterium quality control product, and belongs to the field of molecular biology. The kit comprises a leptospira specific primer probe group, a hot-regressive spirospira specific primer probe group, a borrelia burgdorferi specific primer probe group and other PCR reaction system components, and can realize high-specificity, high-sensitivity and high-accuracy quantitative detection of target genes of the leptospira, the hot-regressive spirospira and the borrelia burgdorferi. The standard requirements of fluorescent PCR and digital PCR multi-gradient standard curve establishment, nucleic acid extraction kit performance verification and clinical diagnosis are met. According to the quality control product, target gene consistency sequences of leptospira, retro-hot spirospira and borrelia burgdorferi are integrated into an escherichia coli artificial chromosome through a homologous recombination technology, the extraction characteristic of a cell structure simulation sample is reserved, the copy number is calibrated through digital PCR, and the quality control product has nuclease degradation resistance.

The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) primer probe and a kit for detecting abalone muscular dystrophy virus. The probe and the primer are designed for a highly conserved region of an AbSV genome, a real-time fluorescent PCR system is established, and the probe and the primer can be used for detection of abalone muscular dystrophy virus and quantitative analysis of virus load; the detection sensitivity is high, and 10 copies / mu L of viruses can be detected; the method is simple to operate, low in detection cost and wide in application range; the detection specificity is high, the detection speed is high, and the detection process can be completed in only 80 minutes.

The invention relates to a quadruple TaqMan real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer group and a probe group for simultaneously detecting cGPV (Complementary Glutathione Virus), MDPV (Minimal Disease Papilloma Virus), MDGPV (Minimal Disease Papilloma Virus) and SBDSV (Sequence Broadcast DSV) and a kit thereof. The sequences of the primer group and the probe group are respectively shown as SEQ ID NO.1-12. The research successfully develops and verifies a multiple TaqMan-MGB real-time fluorescent PCR method capable of simultaneously detecting and distinguishing four important waterfowl parvoviruses (cGPV, MDPV, MDGPV and SBDSV). The method has good specificity, repeatability and high sensitivity. Compared with a conventional PCR method, the multiple detection method has the advantages that the clinical waterfowl parvovirus detection rate is remarkably improved, mixed infection can be effectively recognized, and the clinical diagnosis efficiency is greatly improved.

This invention relates to the field of molecular biology detection technology, specifically disclosing a quadruple fluorescent PCR primer-probe combination, reaction system, and detection method for identifying wild-type and vaccine strains of Mycoplasma gallisepticum. The quadruple fluorescent PCR primer-probe combination for identifying wild-type and vaccine strains of Mycoplasma gallisepticum includes a universal primer-probe set for detecting Mycoplasma gallisepticum and specific primer-probe sets targeting three vaccine strains, F-36, TS-11, and 6 / 85, respectively. The quadruple fluorescent PCR primer-probe combination, reaction system, and method of this invention for identifying wild-type and vaccine strains of Mycoplasma gallisepticum can simultaneously detect and differentiate the DNA of wild-type Mycoplasma gallisepticum and its three different vaccine strains (F-36, TS-11, and 6 / 85), making detection and differential diagnosis more efficient, economical, and convenient; it has advantages such as high sensitivity, good specificity, rapid high throughput, and simple operation.

The invention discloses a multiple real-time fluorescent PCR (polymerase chain reaction) primer probe combination, a kit and a detection method for detecting four common animal-derived components in a blood product. The multiplex real-time fluorescent PCR detection system comprises four groups of primer probe combinations of ducks, chickens, pigs and geese aiming at cytochrome b genes, and the sequences of the four groups of primer probe combinations are sequentially shown as SEQ ID No.1-12. According to the present invention, by using the specificity of the target gene, the primer probe combination only performs specific amplification on the target gene segment of the respective species, and is not interfered by other species; the method has the advantages of high throughput, strong specificity, high sensitivity and the like, the detection limit can reach 0.01% (w / w), and the method is suitable for large-scale rapid detection of four common derived components of ducks, chickens, pigs and geese in blood products.

The invention discloses a hereditary thrombophilia gene detection kit and application thereof, and relates to the technical field of gene detection. The kit comprises three tubes of PCR reaction liquid, and each tube of PCR reaction liquid contains a specific primer and a fluorescent probe and covers 11 SNP sites. According to the hereditary thrombophilia gene detection kit and the application thereof, high-throughput typing of 11 SNP loci is realized through a multi-channel fluorescent PCR and melting curve analysis technology. The kit has the advantages of simplicity and convenience in operation, high sensitivity, low cost and the like, and is suitable for clinical early screening and precise intervention.