244 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 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 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 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 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 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.

The invention relates to the technical field of molecular breeding, and discloses a functional marker development method based on a rice anthocyanin synthesis key gene OsTTG1 and application of the functional marker development method in molecular assisted breeding. The marker comprises a specific primer composition: a forward primer as shown in SEQ ID NO: 1, a reverse primer (used for specifically amplifying an A allele) with the 3'end being AT 'as shown in SEQ ID NO: 2, a reverse primer (used for specifically amplifying a G allele) with the 3' end being AC 'as shown in SEQ ID NO: 3, and universal fluorescent primers as shown in SEQ ID NO: 4 and SEQ ID NO: 5. The invention further provides a kit containing the primer composition, a method for carrying out fluorescent PCR typing by utilizing a PARMS technology and application of the kit in screening high-anthocyanin rice germplasm. The marker is used for directly detecting a functional site, the verification accuracy rate in 419 germplasm reaches 99.05%, the breeding efficiency can be remarkably improved, and the breeding period can be shortened.

The present application relates to the field of virus detection technology, specifically disclosing a dual microfluidic fluorescent PCR detection kit for the H3N2 subtype of avian influenza virus. The dual microfluidic fluorescent PCR detection kit comprises a sample lysis extract for detecting the H3N2 subtype of avian influenza virus, a sample wash solution I, a sample wash solution II, and a reaction premix. The reaction premix contains three sets of specific sequence combinations: an H3-specific sequence combination, an N2-specific sequence combination, and an internal standard gene-specific sequence combination. By designing specific primers and probes, and performing optimization and screening, the present application enables simultaneous identification of H3 and N2 subtype AIVs in a single reaction. Furthermore, the amplification reactions of the two target genes are free of interference, resulting in high specificity, high sensitivity, and good reproducibility. This kit enables accurate diagnosis of viral infection and has important application value in the prevention and control of avian influenza epidemics.

The invention discloses a primer probe composition for detecting measles, parotitis, rubella and varicella virus nucleic acid and application, and belongs to the technical field of virus nucleic acid detection. The invention protects the real-time fluorescent PCR detection primer and probe composition for simultaneously or respectively detecting measles, parotitis, rubella and varicella viruses and different subtype strains, and plays a positive role in respectively or simultaneously performing clinical detection and prevention and diagnosis on different strains of several viruses. The detection method disclosed by the invention is simple, time-saving and labor-saving, wide in application range and convenient to use, and measles, rubella, parotitis and varicella viruses and different strains can be rapidly detected at the same time by amplifying a PCR reaction once. The primer probe composition provided by the invention is high in detection sensitivity, strong in specificity and good in repeatability, can realize rapid differential diagnosis on clinical samples, enables different patients to obtain timely targeted prevention and treatment, and has important significance for development of quadruple vaccines.

The invention provides a nucleic acid composition and a kit for identifying a rice variety rice flower No.2, and relates to the technical field of biochemical detection. The nucleic acid composition comprises a specific amplification primer pair and a specific detection probe. The nucleic acid composition locks the specific SNP site of the rice, and by utilizing the synergistic effect of the primer and the double probes, the rice flowery No.2 variety and the similar variety can be accurately distinguished, and the doping can be sensitively identified. The method is suitable for a real-time fluorescent PCR platform, electrophoresis and sequencing are not needed, rapid and high-sensitivity closed tube detection is achieved, and an efficient means is provided for authenticity identification and purity analysis.

The invention discloses a primer, a TaqMan probe and a kit for carrying out fluorescent PCR (Polymerase Chain Reaction) detection on bovine-derived ureaplasma, and belongs to veterinary detection of animal epidemic diseases in the technical field of biological detection. The invention discloses a primer and a TaqMan probe which are used for carrying out fluorescent PCR (Polymerase Chain Reaction) detection on bovine-derived ureaplasma, and the primer and the TaqMan probe are an upstream primer U.diversum-F3, a downstream primer U.diversum-R3 and a TaqMan probe U.diversum-P3. The kit is simple and convenient in detection operation, strong in specificity, high in sensitivity and good in repeatability, and can be used for qualitatively detecting the bovine-derived ureaplasma; technical support is provided for detection of bovine-derived ureaplasma, epidemic situation epidemiological investigation, epidemic disease monitoring and purification, strain screening and quality monitoring and stable production and supply conservation in the vaccine production process, and the method plays an important role.

The invention discloses a real-time detection system and a monitoring and early warning method for African swine fever virus. The system comprises a latex microsphere labeled immunochromatography antibody detection card, a portable MINI-PCR instrument and a freeze-dried microsphere type fluorescent PCR detection reagent, the detection card takes the recombinant ASFV p30 protein as a target spot, antibody detection is completed within 10 minutes, the lowest detection limit is 1: 1024, and the specificity is greater than 99%; the PCR reagent adopts a double-primer double-probe design, and the lowest detection limit is 1 copy / microliter. According to the invention, through a collaborative process of antibody screening, nucleic acid detection and result evaluation, ASFV total infection cycle detection is realized, epidemic situation discovery time is advanced to 1-3 days after infection, and early warning response time is shortened by more than 60%; the system is simple to operate and low in cost, does not need professionals and laboratories, can be deployed in basic-level scenes such as pig farms and slaughter houses, effectively solves pain points of an existing detection system, and provides key technical support for prevention and control of African swine fever.

This invention discloses a primer and probe set for detecting goat type C Kubuvir based on a temperature-controlled isolated fluorescent PCR platform. It includes upstream and downstream primers for goat type C Kubuvir and a fluorescent probe. This invention also provides a kit for detecting goat type C Kubuvir based on a temperature-controlled isolated fluorescent PCR platform and its application. This kit, used in conjunction with a temperature-controlled isolated PCR instrument, offers rapid detection, high specificity and sensitivity, and is easy to carry and store. It enables rapid and efficient on-site detection of goat type C Kubuvir, solving the technical problems of low detection rate, inability to rapidly and accurately detect various goat type C Kubuvir variants, and inconvenient operation in existing technologies. It shows promising application prospects.

The invention discloses a primer group, a kit and a detection method for high-risk clone typing detection of klebsiella pneumoniae ST11, the primer group comprises one or more amplification primer pairs and probes for detecting high-risk clone marker genes (including bmr3, sdaC, pyrB, mltC and ppsC) of klebsiella pneumoniae and ST11, and the high-risk clone typing detection of klebsiella pneumoniae and ST11 is realized by using a method of asymmetric fluorescent PCR amplification and melting curve. According to the present invention, the mutation type sample and the wild type sample are subjected to the differentiated detection so as to form the stable PCR reaction kit system, such that the clinical klebsiella pneumoniae pathogen can be efficiently and accurately detected, and the ST11 typing detection can be performed on the ST11 high-risk cloning marker gene. A complete system formed by the primer group, the kit and the detection method has high specificity and accuracy, klebsiella pneumoniae pathogens and ST11 high-risk clone typing can be effectively detected, rapid, efficient and low-cost detection can be realized, the detection time is greatly shortened, and large-scale application is facilitated.

The application provides a primer probe set, a kit and application for quantitatively detecting murine infectious agents, and belongs to the technical field of genetic engineering.The six-reagent quantitative detection kit (fluorescent PCR method) for tsutsugamushi oriental body, fever with thrombocytopenia syndrome virus, prrillonella, hantavirus, leptospira, and rickettsia mooseri can detect the six pathogenic targets of tsutsugamushi oriental body, fever with thrombocytopenia syndrome virus, prrillonella, hantavirus, leptospira, and rickettsia mooseri in one reaction, and accurately quantitatively analyzes each pathogenic target.Through single-tube reaction, parallel detection and quantification of the six pathogenic targets are realized, the time consumed due to misjudgment of the targets is reduced, the consumption of consumables and the occupation of instrument space caused by multi-tube reaction are reduced.The quantitative results reflect the pathogenic load, and provide efficient detection and application support for rapid identification and quantitative monitoring of samples of murine origin or samples of the environment related to mouse activities.

The invention relates to a specific real-time fluorescent PCR (polymerase chain reaction) amplification primer, a probe and a detection method for identifying lutjanus purpureus, and belongs to the technical field of molecular detection and species identification. According to the invention, the specific primer and the probe for lutjanus purpuratus are designed, and the real-time fluorescent PCR technology is combined, so that the lutjanus purpuratus can be quickly and accurately identified. The method has the advantages of high specificity, high sensitivity, simplicity and convenience in operation and the like, and a reliable molecular biological method is provided for species identification of lutjanus purpurea.

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