45 results about "Dot blot" patented technology

We describe examples using aptamers for capturing and reporting the presence of a target, such as a pathogen. Examples described here include a set of aptamers that are specific to F. tularensisis. Other examples described here include an Aptamer-Linked Immobilized Sorbent Assay (ALISA) and dot blot assay. An example of a method provided here comprises: providing a set of DNA sequences that exhibit high binding affinity to target antigen, placing the DNA sequences in a sandwich aptamer-linked immobilized sorbent assay (ALISA), contacting the DNA sequences with a sample, and detecting whether the target is present in the sample. Some alternative implementations may include dot blots and different reporters. Quantum dot sandwich assays and quantum dot de-quenching reporters can be used.

The present invention disclosed the use of Allele-Specific-Oligonucleotide (ASO) as the detection assay for human HLA classification. Using the Reversed-Dot-Blotting format and the Flow Through Hybridization process, more efficient, fast and less expensive HLA classification can be achieved. The simple procedures for the process were described. This invention can also provides the method for genotyping as well as DNA analyses in general for different genes and different organisms.

The present invention disclosed the use of single nucleotide polymorphism (SNP) as the detection assay for human identification. Using the reversed dot-blot format and the flow through hybridization process, the process can be more efficient, less expensive and with similar or better power of exclusion in definitive identification. The present method can be applied to any other organisms.

The invention provides a high-specificity and high-sensitivity coordination combination anti-human TK1 antibody prepared from an antigenic determinant consisting of 23 peptide at an N end, 20 peptide at a C end and 28 peptide at the C end of human cervical cancer cells, and application thereof to tumor diagnosis. The antigenic determinant comprises the following amino acid sequences: 1) the 23 peptide (3-25) at the N end: CINLPTVLPGSPSKTRGQIQVIL: 2) the 20 peptide (206-225) at the C end: CPVPGKPGEAVAARKLFAPQ; and 3) the 28 peptide at the C end: AGPDNKENCPVPGKPGEAVAARKLFAPQ. The invention simultaneously provides a method applying the antigen for preparing the antibody. An antibody kit provided by the invention has the characteristics of high sensitivity, high specificity, low cost and the like, tumor rehabilitation populations are periodically monitored by an enhanced chemiluminescence dot blot detection method, immunohistochemistry detection and the detection kit, and the recurrence and transfer risk and the prognosis of the tumor rehabilitation people are evaluated.

The present invention is directed to novel assays for detecting target molecules. The assays employ small size, detectably labeled, magnetic nanoparticles associated with a capture molecule. The detection assay is accelerated by applying magnetic field during the assay. The assays of the invention can be used to enhance the efficiency of the detection step in dot blot, Western blot and ELISA.

Methods and compositions for identifying novel genes that share regions of homology with known genes from target groups of genes of interest are provided. The methods comprise systematically designing oligonucleotide primers that are specific for regions of homology within the nucleotide sequences of a target group of known genes and performing successive rounds of PCR amplification of nucleic acid material from an organism of interest. The PCR steps are intended to identify and amplify nucleic acids comprising both known and novel genes. Nucleic acid molecules comprising known genes are detected and eliminated from further consideration by dot blot analysis using oligonucleotide probes specific for the known genes in the target group. Potentially novel genes are subjected to further sequence analysis to confirm novelty and assayed for biological activity. Compositions of the present invention include novel polynucleotides, and variants and fragments thereof, that comprise novel genes and the polypeptides encoded thereby.

The present invention discloses a nucleic acid molecular hybridization detection method of domestic silkworm nosema disease, including the following steps: using nucleic acid molecular hybridizatino technology to extract DNA of domestic silkworm microsporidian, utilizing PCR to clone the PCR product into recombinant vector plasmid, then using N.P.PCR product and DIG marker to make molecular probe, and adopting DNA Dot blot to detect the pathogenic spore of silwkorm nosame disease. The various microsporidian genomes extracted by said invention are good in quality, high in content, and the designed primers are specific for N.B spora, and said primer and template DNA specificity are highly matched, and its detection accuracy is greater than 98%.

The invention provides a dot blot hybridization reaction device and application thereof, and an immunoblotting detection method and relates to the technical field of molecular hybridization experimental devices. The dot blot hybridization reaction device comprises a sampling infiltration module and a suction filtration module used for supplying a negative pressure environment for the interior of the sampling infiltration module, wherein the sampling infiltration module comprises a micropore sampling plate provided with a sample through hole, a support layer provided with an infiltration through hole corresponding to the sample through hole, and a suction filtration tank; a first actuation unit is arranged in the micropore sampling plate; a second actuation unit matched with the first actuation unit is arranged in the suction filtration tank; a closed inner chamber is formed by the matched actuation units in the manner of adsorbing and fixing; and a hybridization film layer and the support layer are successively fixed under the micropore sampling plate. The dot blot hybridization reaction device is portable, is simple in operation and can effectively prevent the problem of cross contamination of sample rooms of the dot blot hybridization reaction device.

The invention provides a novel serology biomarker GSK3beta detection method for cognitive impairment of a diabetic. The method comprises an enzyme activity assay method and a relative quantification dot-blot method. According to the enzyme activity assay method, a GSK-3beta enzyme activity assay kit of GENMED is applied; GSK-3beta phosphorylation is performed under the suppression of GSK-3alpha; along with oxidation reaction of reduced form of nicotinamide-adenine dinucleotid (NADH), a pyruvate kinase and lactic dehydrogenase continuous circulation method reaction system adopts the spectrophotometric method to measure the peak value change after oxidization so as to reflect GSK 3beta activity in a sample. The dot-blot method transfers blood platelet protein to an NC film according to the antigen and antibody fixation reaction principle and then utilizes antibody to perform detection. The novel serology biomarker GSK3beta detection method is used for studying GSK-3beta protein and enzyme activity expression in the diabetic blood and is expected to become a serology biomarker for early diagnosis of Alzheimer's disease (AD).

The invention provides a processing method for low-allergenic crabs based on directional modification for key amino acids and relates to the field of food processing. The processing method comprises the following steps: salting pretreatment of crabs; directional modification for key amino acids; product packaging. After crabs are cooked in different ways, crude proteins are extracted; SDS-PAGE electrophoresis and Dot Blot are adopted for analyzing the crude proteins after thermal processing at different degrees; the allergenicity of crabs under different processing modes is judged according tothe degree of immune binding on the basis of taking binding activity of a rabbit anti-TM polyclonal antibody as an index; the processing mode of low-allergenic crabs is screened. Simple antigen antibody combination reaction is utilized to confirm the change in allergenicity after crab allergen is subjected to different thermal processing; the processing method for low-allergenic crabs based on directional modification for key amino acids is screened; the process is simple, the operability is high and the processing method is easy for large-scale production.

The invention discloses a method for identifying iron deficiency-resistant apple rootstocks. The method comprises the following steps: 1) conducting iron deficiency treatment on an apple rootstock to be detected and a standard iron deficiency-resistant apple rootstock respectively, and extracting total RNA; 2) conducting dot-blot hybridization on the total RNA with an iron deficiency-resistant gene probe; 3) scanning with an image scanner, and conducting an analysis with an image master to obtain the expression quantities of the iron deficiency-resistant genes in the apple rootstock to be detected and the standard iron deficiency-resistant apple rootstock; and 4) if the expression quantity of the iron deficiency-resistant genes in the apple rootstock to be detected is not lower than that of the iron deficiency-resistant genes in the standard iron deficiency-resistant apple rootstock, determining that the apple rootstock to be detected is an iron deficiency-resistant apple rootstock or an iron deficiency-resistant apple rootstock candidate. The method provided by the invention has the advantages of low cost and high efficiency, overcomes the limitation of conventional selective breeding of iron deficiency-resistant apple rootstocks, conducts selective breeding at the gene level, and lays a good foundation for selective breeding with the aid of gene expression marking.

Provided is a method for quantitative analysis of a sample. The method includes steps of (a) providing a singular marker representative of one or more features of the sample, the sample comprising a population of individual units of the marker; (b) measuring the marker with dot blot analysis, wherein the quantitation result is an absolute amount of the marker's population of individual units in the sample, normalized by the sample volume or by the sample weight; and (c) obtaining an objective determination of the one or more features of the sample based on the quantitation result of the marker. Also disclosed is a reference database and a method to use the reference database for diagnosing cancer in a patient.

The invention relates to kendir DNA untwisting enzyme gene APDH1 clone, recombination, salt tolerance function analysis, and transgene salt and alkali proof cotton breeding selection. It belongs to molecular biology and biotechnology field. It includes the following steps; utilizing inhibition difference subtraction crossing technique to form kendir DNA positive direction difference subtraction cDNA library; processing enzyme cutting for positive cDNA clone, PCR and reverse direction Northern dot blot hybridization identification, DNA sequencing and nucleotide sequence homology comparison to gain cDNA sequence; one segment of the sequence is DNA untwisting enzyme gene middle segment; processing 3í»and 5í»end fast amplification to gain coding kendir DNA untwisting enzyme span cDNA named APDH1; and forming expression vector to transform cotton. The formed transgene cotton plant can normally come out, flower, and boll opening at 0.45% alkaline land.

The present invention disclosed the use of single nucleotide polymorphism (SNP) as the detection assay for human identification. Using the reversed dot-blot format and the flow through hybridization process, the process can be more efficient, less expensive and with similar or better power of exclusion in definitive identification. The present method can be applied to any other organisms.

The invention relates to a gene chip, an amplification reagent and a kit for detecting alpha-thalassemia, and belongs to a molecular diagnosis technology, wherein 3 non-deletion (alpha<CS>alpha, alpha<QS>alpha and alpha<WS>alpha) alpha-thalassemia and 4 deletion (--<SEA>, --<THAI>, -alpha<4.2> and -alpha<3.7>) alpha-thalassemia can be rapidly and simultaneously detected by combining direct PCR andreverse dot blot (RDB).

The invention provides a probe and kit for detecting fetal free DNA beta-thalassemia mutation in maternal blood. The kit comprises a detection probe, a capture probe, a reaction film strip, a target gene amplification reagent, a cleaning solution, an incubation solution and a developing solution. According to the kit, a dot blot hybridization method is utilized; in the method, a DNA hybridizationprocess is used for detecting a product; and the kit has the advantages of simplicity in operation, rapidness, simplicity, convenience, small sample dosage, intuitive and reliable result and the like,and can be widely applied to detection of fetal free DNA beta-thalassemia mutation in maternal plasma in noninvasive prenatal diagnosis.

The invention relates to tumor markers, in particular to the application of prothymosin alpha to preparation of a breast cancer diagnosis marker. According to the application of prothymosin alpha to preparation of the breast cancer diagnosis marker, the dot blot hybridization method and the ELISA method are applied to detection of the level of prothymosin alpha protein in urine of a breast caner patient, and the relation between prothymosin alpha and the breast cancer biological behaviors is discussed; the immunoassay method is adopted, it is only required that the level of prothymosin alpha in urine is detected, it can quickly prove that the level of prothymosin alpha in urine of the breast cancer patient is obviously higher the normal level, preliminary judgment is conducted on breast cancer, and a basis is provided for further diagnosis; the needed urine sample is easy to obtain, any wound is avoided, an expensive instrument is not needed, operation is convenient, and popularization and application are easy.

The invention discloses a mycobacteria strain detecting kit, comprising a dot blot membrane. A specific oligonucleotide probe for specifically detecting 23 mycobacteria and a primer for specifically amplifying an insertion sequence between mycobacteria 16S rRNA and 23S rRNA are fixed. The mycobacteria strain gene detecting kit adopts PCR combining with reverse DNA hybrid chip detection technology to simultaneously identify 23 mycobacteria within one day. The novel technology can simultaneously identify the most mycobacteria within shortest time and has the most reliable detection result. The kit distinguishes mycobacteria through the difference of gene sequences of different mycobacteria and carries out distinction and identification in the level of gene sequence of bacteria and molecular structure, so that the classification is more accurate and reliable.

We describe examples using aptamers for capturing and reporting the presence of a target, such as a pathogen. Examples described here include a set of aptamers that are specific to F. tularensisis. Other examples described here include an Aptamer-Linked Immobilized Sorbent Assay (ALISA) and dot blot assay. An example of a method provided here comprises: providing a set of DNA sequences that exhibit high binding affinity to target antigen, placing the DNA sequences in a sandwich aptamer-linked immobilized sorbent assay (ALISA), contacting the DNA sequences with a sample, and detecting whether the target is present in the sample. Some alternative implementations may include dot blots and different reporters. Quantum dot sandwich assays and quantum dot de-quenching reporters can be used.

The invention discloses an autoimmunity detection kit with enzyme-linked immuno spot blotting and an operation process thereof. The autoimmunity detection kit with the enzyme-linked immuno spot blotting comprises a kit body, target molecules and a reagent. The kit is a technology based on combination of enzyme-linked immuno spot blotting and enrichment and is used for detecting pathoglycemia caused by deficiency of the autoimmunity function. The operation process of the autoimmunity detection kit with the enzyme-linked immuno spot blotting comprises the following steps of: preparing the targetmolecules, preparing a covering device of the target molecules, preparing cell lysis solution, preparing anti-human immune cell antibodies, enriching the immune cells, preparing amplification culturemedium, detecting the target molecules and analyzing clinical-sample detection data. The autoimmunity detection kit and the operation process disclosed by the invention have the beneficial effects that by combination of the immune cell enrichment technology and the enzyme-linked immuno spot blotting technology with high sensitivity and high specificity and accurate analysis for the correspondingtarget molecules, the detection specificity is improved and the false positive probability is reduced.

The invention is a procedure for measuring the binding of an entity (ligand) to a surface by using a hapten-conjugated version of the ligand (hapten-ligand). An excess of the hapten-ligand is presented to the binding surface and excess (unbound) hapten-ligand is washed off. Bound hapten-ligand is then solubilized (removed) and applied to a membrane support or separated by electrophoresis and applied to a membrane support. Known amounts of hapten-ligand are similarly applied to the membrane, to provide for hapten-ligand standards. The membrane-bound hapten-ligand is detected by application of an enzyme-conjugated antibody to the hapten; or by application of an antibody to the hapten followed by application of an enzyme-conjugated antibody to the anti-hapten antibody. The resultant membrane-associated enzyme is detected and quantitated by the application of a color or light-producing substrate which reacts with the enzyme. A combination of the use of anti-hapten antibodies along with membrane-blotting technologies to assess hapten-ligand binding to surfaces is not found in the scientific or patent literature, particularly in regards to assessing protein binding to cell surfaces.

The invention relates to the technical field of marine organisms and in particular relates to a method for rapidly screening a microorganism strain capable of generating tetrodotoxin and a digoxin labeled DNA (Deoxyribonucleic Acid) probe used by the method. According to the method provided by the invention, the utilized probe is single-stranded DNA with a length of 254 basic groups which are complementary with a DNA basic group to be detected, and a 3' terminal of the probe is labeled by digoxin. Colony in-situ dot blot molecular hybridization is carried out, and the probe can be used for specifically detecting whether a strain DNA sample to be detected has tetrodotoxin synthesizing sxt gene nucleic acid or not, and a toxin-producing positive strain is screened. According to the method provided by the invention, DNA in-situ extract of the strain is used for carrying out colony dot hybridization, and 96 samples can be screened within 8 to 10 hours. The method provided by the inventionis rapid, high in specificity and high in accuracy.

The invention provides whole blood RNA quick lysate which does not need erythrocyte pyrolysis firstly, and an RNA extraction method. According to the method, the usage of toxic reagents is reduced toa large extent, RNA extraction steps are simplified, and RNA obtained through purification is good in integrity and high in purity and can be directly used for various molecular biology experiment ofRT-PCR, Northern blot, Dot blot and the like. Every 1000ml by volume of the whole blood RNA quick lysate disclosed by the invention consists of the following components by dosage of 0.2M-4M of guanidine thiocyanate, 0.2M-3M of ammonium thiocyanate, 0.01M-0.2M of anhydrous sodium acetate, 0.05M-0.3M of glacial acetic acid, 0.1%-2%(W / V) of sodium lauroyl sarcosine, 0.02M-0.3M of sodium chloride, 1%-15%(V / V) of glycerine, 10%-75% (V / V) of a water saturation phenol solution, and the balance DEPC treating water.

The invention relates to a method for detecting the infection of avian anemia viruses in a pathologic material by the combination of polymerase chain reaction (PCR) and a nucleic acid probe and dot blot hybridization technology. According to the method, deoxyribonucleic acid (DNA) of a tissue sample of an animal (dead animal) which is suspected to be infected with the avian anemia viruses is subjected to PCR amplification by using a specific primer for the avian anemia viruses, and then a PCR product is subjected to dot blot hybridization by using a specific nucleic acid probe for the avian anemia viruses, so that the specificity of the PCR product can be shown, and the detection rate is also greatly improved. The experimental result shows that: the infection of the avian anemia viruses in the pathologic material is detected by the combination of the PCR and the nucleic acid probe and dot blot hybridization technology, so that the sensitivity and specificity of the detection of the infection of the avian anemia viruses can be remarkably improved.

The present invention provides methods of unambiguously identifying a human herpesvirus in a sample. The assays, which allow for the detection and typing of all ten human herpesviruses, involve multiplex PCR assays using consensus primers to amplify conserved regions of the herpesvirus DNA. A dot blot / chemiluminescence assay and real time PCR assay ideal for clinical setting were disclosed. A heteroduplex mobility assay suitable for uses in research laboratory was also presented.

The invention provides a reaction cup. The reaction cup comprises a cup wall and a cup bottom; the cup wall is further provided with retaining parts; the retaining parts can make dot blot membranes stand upright in the reaction cup, and a dot blot membrane upright placement position is formed in a region of connecting lines of two opposite retaining parts. The invention further provides a nucleicacid molecular hybridization instrument comprising the reaction cup, and a detection method for detecting an analyte. The new reaction cup provided by the invention can vertically hold the dot blot membranes, and can be operated in an open state, so that the area of the reaction cup on the molecular hybridization instrument is reduced; and more reaction cups can be placed on the molecular hybridization instrument once, so that the working efficiency of the molecular hybridization instrument to detect the analyte is improved.

The invention discloses a Digoxin probe for human-derived MANF gene detection and application thereof. The sequence of the Digoxin probe is shown as SEQ NO:1. The invention further relates to the application of the Digoxin probe for human-derived MANF gene detection in a recombinant virus titer detection method. The invention further relates to a dot blot hybridization detection method which involves the Digoxin probe in claim 1. The Digoxin probe for human-derived MANF gene detection is creatively designed for the first time in at home and has high specificity. The designed Digoxin probe can be applied to relevant recombinant virus titer detection, has high detection specificity and sensitivity and brings great convenience to relevant experimental study.

The invention discloses a method for jointly detecting a whole genome DNA adduct based on dot blot hybridization and chromatin co-immunoprecipitation sequencing. The method adopts the dot blot hybridization technology to detect the total amount of the DNA adduct and optimize each test condition so as to obtain the DNA adduct state of the whole genome in the maximum range, then the chromatin co-immunoprecipitation sequencing technology principle is introduced, a DNA fragment library of a BPDE-DNA adduct is generated in an established contaminated cell by using a specific recognition reaction of an antigen and an antibody, an injury map is obtained by a sequencing technology, the site of the DNA adduct in a whole genome is defined so as to screen a gene related to lung cancer caused by BPDE, and the gene related to lung cancer caused by BPDE is screened by combining a BPDE-DNA adduct antibody and specifically enriching a DNA fragment acted by BPDE for sequencing, so that more comprehensive and exact information is provided for further screening key pathways and targets of B[a]P carcinogenesis.

The invention relates to a monoclonal antibody B of the pathogenic toxin protein PirA of the prawn hepatopancreas and its application. The monoclonal antibody B is secreted by a hybridoma cell with the preservation number: CCTCC NO: C2017153, and can interact with the epitope of the toxin protein PirA specific binding. The application is to apply the monoclonal antibody B to the pathogen detection of acute hepatopancreatic necrosis in prawns, specifically, to detect whether the prawns contain PirAB plasmid pathogens by using the dot blot method. The monoclonal antibody B developed by the present invention can combine with the PirA protein epitope, so that the monoclonal antibody B can be applied to the early rapid detection of the AHPND pathogen, in order to establish a monoclonal antibody diagnostic method for the AHPND pathogen, and to study the pathogenesis of AHPND It has established a solid foundation for cutting off the infection route, preventing and controlling shrimp AHPND through the infection dynamics, proliferation characteristics, and epidemiological laws.

The invention belongs to the technical field of animal pathogen molecule detection, and particularly relates to a detection primer, a detection kit and a virus detection method for a fowl adenovirus-I, and application of the primer. A specific primer is used for synthesizing a digoxigenin-labeled specific nucleic acid probe through PCR (Polymerase Chain Reaction). Through dot blot, the specific probe can be used for detecting the existence of 12 types of serotype nucleic acids of the fowl adenovirus-I in a pathological material sample, and is used for judging whether fowl adenovirus-I infection is in the presence or not. By use of the kit to which the invention relates, the DNA (deoxyribonucleic acid) of a pathological material tissue sample is extracted to carry out the dot blot, detection can be finished in 24-36h, a result is reported, so that specificity is high, and accuracy is high.