198 results about "Nucleic acid test" patented technology

This invention describes a rapid (10 to 15 minutes), simple, flexible and efficient method of nucleic acids extraction for nucleic acid testing assays. This method has the following basic steps: i) mechanical cell lysis using solid particles in the presence of a chelating agent, followed by ii) controlling the presence and / or activity of NAT assays inhibitors. This method is applicable to various biological samples and universal for microorganisms, as one can use it to extract nucleic acids from test samples containing target viruses, bacteria, bacterial spores, fungi, parasites or other eukaryotic cells, including animal and human cells.

The invention discloses a one-step nucleic acid test method based on CRISPR / Cas (clustered regularly interspaced short palindromicrepeats / CRISPR-associated protein) and isothermal amplification and akit. The method is a quick field test method used for testing nucleic acid and displaying visual test results through fluorescent light or lateral side immunochromatographic test paper. The test accuracy is effectively ensured through specificity of crRNA (RISPR-derived ribonucleic acid); at a room temperature, whether a test sample contains test nucleic acid can be determined through the test paper or the fluorescent light within 1-2h; and compared with the traditional test method, the precision, simplicity and convenience in operation of the test method are improved to some extent. The kit with purified freeze-drying LbaCas12a, crRNA and RPA (recombinase polymerase amplification) can be formed; a nucleic acid segment of a specific sequence can be conveniently and quickly tested on site;a complicated temperature control instrument such as a PCR (polymerase chain reaction) instrument and other electrophoresis and centrifugation equipment are not required; and nucleic acid of the specific sequence can be tested quickly and sensitively only by a thermostatic device and fluorescence detection equipment and even visual observation.

The invention discloses a method for detecting food-borne pathogenic bacteria by using a colloidal gold nucleic acid test strip based on hyper-branched rolling cycle amplification and a kit for detecting Listeria monocytogenes and Salmonella, belonging to the technical field of biological detect. The method comprises the following main steps: (1) performing extraction and enzyme digestion on the DNA (deoxyribonucleic acid) of food-borne pathogenic bacteria; (2) designing an amplification primer and a nucleic acid probe sequence; (3) performing hyper-branched rolling cycle amplification reaction; (4) annealing and hybridizing; (5) preparing a nano gold probe; (6) preparing a colloidal gold nucleic acid test strip; and (7) detecting a sample. When the sample contains a target gene segment, both a T line and a C line of the colloidal gold nucleic acid test strip are formed into red lines; and if the sample does not contain the target gene segment, only the C line of the colloidal gold nucleic acid test strip is formed into a red line. According to the invention, food-borne pathogenic bacteria in the food sample can be detected quickly, specifically, sensitively, qualitatively and quantitatively; and the invention is simple in probe design, short in operation steps and favorable for popularization.

The invention relates to a novel method for isothermal amplification detection of a double-stranded nucleic acid based on nicking enzyme, belongs to the field of nucleic acid test, and in particular to a method for amplifying a single-chain target from a double-stranded nucleic acid target under the combined action of the nicking incision enzyme and polymerase and carrying out efficient specificity amplification on the target nucleic acid employing two amplification products and a replacement primer in an isothermal condition. Through selection of one amplification primer and an amplification template annealing region, efficient specificity amplification of the target nucleic acid can be finished by only three primers; the 3' end of the template is completely complementary with the primer; the specificity is improved; meanwhile, the problem of refractory amplification due to the fact that the replacement primer occupies the complementary position of the amplification primer in the prior art is avoided; the amplification primer can also be designed into a form of a molecular beacon; and only correctly amplified target sequence can be annealed together with the primer in the form, and generates subsequent fluorescence signals, so that the method has relatively good specificity.

The invention relates to the technical field of gene detection, in particular to a nucleic acid detection method based on CRISPR / Cas and nucleic acid test paper and a human papilloma virus rapid detection kit. The human papilloma virus detection kit comprises crRNA, Cas12, RPA upstream and downstream primers, a buffer system and a single-stranded DNA reporter molecule, the sequence of the reportermolecule is 5 '-NNN......-3', the length of the reporter molecule is 3-30 basic groups, FAM, DIG and Biotin labeling is carried out, and meanwhile modification for preventing DNA enzyme or cas12 degradation after activation is carried out on nucleic acid molecules. The CRISPR / Cas and nucleic acid test paper-based nucleic acid detection method and the human papilloma virus rapid detection kit havethe advantages that innovative reporter molecule design and modification are adopted, and corresponding test paper strips are combined, so that non-specific color development is completely eradicated, and the kit has remarkable significance in clinical practical application.

The invention relates to a method for detecting staphylococcus aureus based on a nucleic acid chromatography biosensing technique. The method provided by the invention comprises the following steps: designing a loop-mediated isothermal amplification (LAMP) primer (SEQ ID NO:1-4) according to a virulence gene nuc of the staphylococcus aureus, and establishing a staphylococcus aureus detection method based on an LAMP nano enzyme sensor by combining with a nano nucleic acid test strip. The method provided by the invention can be successfully used for distinguishing viable bacterial cells and dead bacterial cells, and lower detection limit on the staphylococcus aureus can reach 10CFU / mL.

The invention belongs to the technical field of molecular biology, immunology and nucleic acid chemistry and in particular relates to a non-amplification nucleic acid hybrid capture system and application thereof. The kit is an immunoassay method for capturing hybrid nucleic acids based on a non-amplification anti-nucleic antibody; inverse transcription, PCR (Polymerase Chain Reaction) and other amplification technologies are not needed in the detection process, while a probe complementary to a to-be-detected nucleic acid is subjected to annealing hybridization with the to-be-detected nucleic acid so as to form a structure which can be recognized by the anti-nucleic antibody, and the detection signal is amplified by utilizing enzyme linked immunosorbent assay. The detection method has the advantages of being rapid, low in cost, high in sensitivity, high in specificity, free in complicated amplification and detection instruments and capable of detecting initial viral load.

The invention provides a nucleic acid test reagent card and relates to the technical field of nucleic acid test equipment. The nucleic acid test reagent card comprises a reagent card body provided with a sample region, an extraction unit, an amplification unit, a test unit, a power unit and a microchannel system; the power unit can be communicated with one of a sample tube arranged in the sample region, the extraction unit, the amplification unit and the test unit; the power unit is used to drive a liquid to flow in the microchannel system. The nucleic acid extraction step, the amplification step and the test step can be completed in one reagent card; operating is simple; multiple devices for operating are not required; switching samplings among the multiple devices is not required either; test time is shortened, and the problem is solved effectively that samples are susceptible to external contamination or samples contaminate the environment easily.

The invention discloses a kit with RT-LAMP nucleic acid test strips for detecting a porcine epidemic diarrhea virus and applications of the kit. The kit comprises a primer group of a nucleic acid represented by SEQ ID No. 1-6 and test strips for detection of nucleic acid. The usage of the kit is as follows: first preparing a RT-LAMP reaction system comprising an AMV retrovirus, a 1* reaction buffer, a strand displacement DNA polymerase, a dNTP mixture, betaine, MgSO4, a FIP primer, a BIP primer, a LoopF primer, a LoopB primer, a F3 primer, a B3 primer and RNA of a sample to be measured; carrying out a reaction at a constant temperature, after testing the obtained products by using the nucleic-acid-detecting test strip, judging and reading directly: the positive result is that two red strips appear, and one strip is in the detection zone while the other strip is in the control zone. The kit has advantages of simple operation, low cost, easy observation of the reaction result, good specificity, easy popularization and application in large scope and being extremely suitable for export quarantine, food hygiene and on-site detection in animal husbandry.

The invention relates to a method for detecting salmonella based on a nucleic acid chromatography biosensing technology. According to a virulence gene invA of salmonella, loop-mediated isothermal amplification primers (SEQ ID NO:1-4) are designed, and through the combination with a nano-enzyme nucleic acid test strip, the salmonella detection method based on an LAMP nano-enzyme sensor is established. The method for detecting salmonella based on the nucleic acid chromatography biosensing technology can be successfully used for distinguishing live bacterial cells from dead bacterial cells, and the lower limit of detection on salmonella can reach 10 CFU / mL.

The invention discloses a primer pair which is used for amplification of a TK gene sequence of Cyprinid herpesvirus III. The invention also discloses an amplification method for the TK gene sequence of the Cyprinid herpesvirus III. The invention discloses a detection kit for the Cyprinid herpesvirus III. The invention discloses a method for detecting the Cyprinid herpesvirus III by using a fluorescent quantitative detection kit. The detection kit and the method can provide assurance for rapid and accurate detection of the Cyprinid herpesvirus III, and provide guarantee for disease prevention, scientific medication and fish health. The detection method and the kit on the basis of a special primer provided by the invention can be used for a nucleic acid test of a TK gene of main diseased tissues (the liver, the brain and the kidney) of fish.

The invention discloses a detection method of gene II type grass carp reovirus. The detection method comprises the following steps: (1) extracting the RNA of the grass carp sample, adding RNA into a premix reaction solution, and performing loop-mediated isothermal nucleic acid amplification on the conserved sequence of the gene II type grass carp reovirus by two pairs of primers, wherein the primers are respectively a primer GCRVII-F3 with the sequence of SEQ ID NO:1, a primer GCRVII-B3 with the sequence of SEQ ID NO:2, a primer GCRVII-F1P with the sequence of SEQ ID NO:3 and a primer GCRVII-BIP with the sequence of SEQ ID NO:4; labeling an FITC probe, wherein the sequence of the probe GCRVII-FITC-Probe is SEQ ID NO:5; and (2) dropwisely adding the amplified product on a sample spotting position of the LFD test paper, dropwisely adding a buffer solution in the front end of the sample position, finishing the reaction after the buffer solution is flown to the other end of the test paper, and judging the detection result according to the color development tape on the test paper. By combining the isothermal amplification and the nucleic acid test strip rapid detection technology, the detection method and the kit provided by the invention have the characteristics of simple operation, high detection reliability, good specificity and high sensitivity.

The invention relates to a method for detecting enterobacter sakazakii based on a nucleic acid chromatography biosensing technique. According to a toxicity gene ompA of the enterobacter sakazakii, a loop-mediated isothermal amplification primer (LAMP) (shown in SEQ ID NO:1-4) is designed, and an enterobacter sakazakii detection method based on an LAMP nano-enzyme sensor is established by combining with a nano-enzyme nucleic acid test strip. The method provided by the invention can be successfully used for distinguishing viable bacterial cells and dead bacterial cells, and lower detection limit on the enterobacter sakazakii can reach 10CFU / mL.

The invention relates to a triple nucleic acid test kit. The kit comprises a thallus lysate, a positive quantitative reaction liquid, a negative control reaction liquid and a thallus test reaction liquid, wherein the thallus test reaction liquid contains specific primers, specific probes and doping fluorescent dye for aspergillus, Cryptococcus neoformans and candida albicans, the three specific oprimers are designed according to areas of ITS sequences of aspergillus, Cryptococcus neoformans and candida albicans, FAM, VIC and CY-5 are selected for mark report groups of the three specific probes, MGB serves as a quenching group, the excitation wavelength of the FAM group is 492 nm, and the emission wavelength is 517 nm; the excitation wavelength of the VIC group is 557 nm and the emission wavelength is 574 nm; the excitation wavelength of the CY-5 group is 646 nm and the emission wavelength is 664 nm. The triple nucleic acid test kit for aspergillus, Cryptococcus neoformans and candida albicans can assist in diagnosing infection caused by aspergillus, Cryptococcus neoformans and candida albicans.

The invention establishes a single-tube multi-primer mini-pool (MP) human immunodeficiency virus (HIV) nucleic acid test (NAT) technology with combination of RNA (ribonucleic acid) reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR, which is applied in men who have sex with men (MSM) and other high-risk groups for detection during the window period.

The invention provides a DNA polymerase with DNA or RNA as a template. The DNA polymerase is obtained through amino acid replacement, including G198W, V222I, E306K, Q354E, A381E and E582K, of klenow fragments of escherichia coli polymerases I. The invention also provides a primer with a stem loop structure and used for constant-temperature nucleic acid amplification. The invention further providesa nucleic acid test technique and a test kit both based on combination between rapid constant-temperature nucleic acid amplification and a Cas test system. The DNA polymerase, the nucleic acid test technique and the test kit can be used for testing nucleic acid targets in test samples at a constant temperature, have the advantages of low cost, short test time, simplicity and convenience in operation, high specificity, high sensitivity and the like, and are particularly applied to POCT (point-of-care testing).

The invention discloses automatic nucleic acid sampling and detecting equipment. The equipment comprises a detection cabin, and a sampling device, a detection device and a control system which are arranged in the detection cabin, wherein the detection cabin comprises a cabin body and a solar power generation system, and an entrance is formed in one side of the cabin body and provided with an intelligent door for opening or closing the entrance; the sampling device comprises a jaw bracket, a multi-degree-of-freedom mechanical arm and an image collector; and the detection device comprises a test tube conveying belt, a sorting mechanical arm, a constant-temperature box, a medicine injector and a color recognizer, wherein the sorting mechanical arm sorts nucleic acid test tubes into the constant-temperature box and sorts the nucleic acid test tubes to the position below the color recognizer for color recognition, and the color recognizer transmits detection information to the control system. The whole nucleic acid detection process does not need the operation of nurses, and the operations from nucleic acid detection, to appointment auditing, automatic sampling of pharyngeal specimens and finally nucleic acid detection result sending, are completed by corresponding devices, so that cross infection during manual detection is avoided.

The present invention discloses a COVID-19 nucleic acid test sampling kiosk having a positive pressure and a disinfection apparatus, comprising a base and a cabinet body fixedly mounted on the base. Barrier glass is provided on at least one side surface of the cabinet body; two through holes are provided on the barrier glass, and are both connected to rubber extension gloves; a glove conveyor anda glove withdrawer are mounted on both sides of the barrier glass, respectively; rolled glove belts are mounted on the glove conveyer and the glove withdrawer by means of rolling shafts; and the barrier glass is provided with roller sealing strips on the upper and lower sides of the rolled glove belts, respectively. By means of a physical barrier, medical workers are protected from being infectedby sprays disseminated from oral cavities and nasal cavities of patients during nucleic acid test sampling. By means of a positive-pressure efficient filter air conditioning system, non-filtered external air is prevented from flowing in to reduce the infection risk of doctors by aerosol and to improve the working temperature in the cabinet body. By means of a disinfectant spray apparatus, gloves and the barrier glass are disinfected before and after each test sampling to reduce the risk of cross infection between the patients.

The invention provides a constant-temperature amplification detection kit for detecting dengue nucleic acids accurately, sensitively and rapidly, and a detection method. The detection kit is suitable for qualitative detection of dengue viruses (I, II, III and VI types). The kit comprises a constant-temperature amplification reaction solution, a positive control and a negative control. First, extracted dengue virus RNAs are subjected to amplification for 90min at a constant temperature of 60 DEG C through a cross primer constant-temperature amplification target nucleotide sequence method. Second, the products after amplification are hybridized with two probes with Biotin and fluorescein isothiocyanate (Fitc) labellings respectively in the constant-temperature amplification reaction solution, and the results are shown on nucleic acid test strips. Finally, detection reports are obtained through contrastive analysis with the positive and negative controls. The kit is advantaged by good specificity, high sensitivity and good repeatability. The kit can finish rapid detection of samples within 2h without complex apparatuses, and can meet detection requirements of high throughput and low throughput at the same time.

The invention relates to a method for detecting escherichia coli O157 based on a nucleic acid chromatography biosensing technology. According to a virulence gene rfb E of escherichia coli O157, loop-mediated isothermal amplification primers (SEQ ID NO:1-4) are designed, and through the combination with a nano-enzyme nucleic acid test strip, the escherichia coli O157 detection method based on an LAMP nano-enzyme sensor is established. The method for detecting escherichia coli O157 based on the nucleic acid chromatography biosensing technology can be successfully used for distinguishing live bacterial cells from dead bacterial cells, and the lower limit of detection on escherichia coli O157 can reach 10 CFU / mL.

The invention relates to a novel nucleic acid test micro-fluidic chip device for automatic extraction and purification detection of nucleic acid. The novel nucleic acid test micro-fluidic chip deviceis of a sealed cavity type structure and comprises a waste liquid chamber, a sample reagent chamber, a mixing chamber, a pre-amplifying chamber, an amplifying chamber, a biosensor and a signal outputport, wherein the waste liquid chamber, the sample reagent chamber, the mixing chamber, the pre-amplifying chamber and the amplifying chamber are connected through fluid microchannels, the biosensor and the signal output port are connected through electrical signals, and the fluid microchannels are distributed in the novel nucleic acid test micro-fluidic chip device in a three-dimensional and tubular mode; micropumps are respectively arranged in the waste liquid chamber, the sample reagent chamber, the mixing chamber and the pre-amplifying chamber, or flow dividing holes which are respectivelyconnected with external micropumps are formed in the waste liquid chamber, the sample reagent chamber, the mixing chamber and the pre-amplifying chamber. According to the novel nucleic acid test micro-fluidic chip device disclosed by the invention, pipelines can be cleaned by the micropumps, circular, repeated and discontinuous detection can be achieved, one item or a plurality of items of detection of one sample or a plurality of samples is achieved, and use cost is reduced.

The invention discloses a RT-LAMP nucleic acid test-strip kit for determining hog cholera virus and application. The kit comprises a primer group with nucleotide sequences shown as SEQ ID NO. 1-6 and nucleic acid detection test strips. The application method of the kit comprises as follows: preparing a RT-LAMP reaction system which comprises AMV retrovirus, a 1* reaction buffer, strand displacement DNA polymerase, a dNTP mixture, betaine, MgSO4, a FIP primer, a BIP primer, a hybridization probe, a LoopB primer, a F3 primer, a B3 primer and RNA of a sample to be measured; and carrying out a reaction at a constant temperature, after testing the obtained products by using the nucleic acid detecting test strip, judging and reading directly: the result is positive when two red bands appear, and one band is in the detection zone while the other band is in the control zone. The kit has the advantages of simple operation, low cost, easy observation of the reaction result, good specificity and easy popularization and application in large scope.

The invention discloses a macrobrachium rosenbergii nodavirus detection kit and detection method. According to the detection kit, with the combination of the characteristics of isothermal amplification and rapid detection of nucleic acid test strips, expensive instrument equipment is not required during the detection process; and the color display is performed by utilizing the nucleic acid test strip, so that the detection reliability and the result judgment easiness are improved. The detection kit is low in detection cost and convenient to use and is safe for people and environment, and is capable of replacing the existing relevant detection methods. The detection kit can be used on wild production site and in laboratories in various levels, and has popularization and application value in the aspects of enhancing macrobrachium rosenbergii nodavirus detection, discovering and controlling the individual propagation of infected macrobrachium superbum infected by virus in real time, discovering and preventing the large-scale epidemic outbreak of the epidemix disease and the like.

The invention discloses a method for quickly identifying a food pathogenic bacteria subtype based on an asymmetric polymerase chain reaction (PCR) combined test strip platform and a kit, and belongs to the technical field of biological detection. The method comprises the main steps of: (1) extracting food pathogenic bacteria subtype DNA; (2) designing an amplification primer and a nucleic acid probe sequence; (3) performing amplification reaction based on asymmetric PCR; (4) preparing a nanogold probe; (5) preparing a colloidal gold nucleic acid test strip; (6) detecting a sample. According to the method, food pathogenic bacteria in food samples can be quickly, specially, sensitively, qualitatively and quantitatively detected and the special subtype of the pathogenic bacteria is determined; the probe is simply designed, the operation steps are simple and short, the product is a single chain, and the method is easy to detect and convenient to popularize.

The invention discloses a nucleic acid isothermal amplification detection kit for Salmonella and a detection method. The detection kit comprises a DNA extraction reagent, an isothermal amplification reaction liquid, a positive control and a negative control. The detection kit is high in specificity and high in sensitivity; the temperature of all nucleic acid isothermal amplification reactions is consistent, only 35 minutes are required for isothermal amplification, 1-2 minutes are required for a detection result of a nucleic acid test strip, only about 1 hour is required for the whole detection process from sample receiving to result acquisition, only one isothermal instrument is required in the whole reaction process, and the detection kit is particularly suitable for on-site and rapid detection and elimination of the Salmonella in food.

The invention discloses a colloidal gold nucleic acid test strip for watermelon bacterial fruit blotches as well as preparation and application thereof, which belong to the technical field of colloidal gold detection. A detected substance is firstly bound with colloidal gold coupled Aptamers (1) in a competitive way under the action of a capillary siphoning effect formed by an absorbent pad, and residual aptamers drift to a detection zone to be bound with streptavidin-Aptamers (2) and realize color development in the presence of excessive coupled Aptamers (1); and the V region binding sites of the colloidal gold coupled Aptamers (1) bound with the detecting substance are occupied by the detected substance, so that the Aptamers can only cross the detection zone and drift to a reference zone to be unspecifically bound with streptavidin-Aptamers (3) at binding sites in a C region and carry out color comparison with the detection zone to semi-quantitatively detect the residual quantity of watermelon bacterial fruit blotches in the sample. The test strip can meet the requirement of food safety on the residual quantity of watermelon bacterial fruit blotches, is suitable for feeds, meat producing plants and government detection mechanisms, and has the characteristics of convenience for using, economical efficiency, rapidness, easiness in manufacturing and low cost.

The invention discloses a cyprinid herpes virus type 2 CPA detection primer and application. A cyprinid herpes virus type 2 CPA detection kit comprises 10*ThermoPol Reaction Buffer, Bst DNA polymerase, dNTPs, a cross primer 1S, probe primers 2A and 3A, stripping primers 4S and 5A, MgSO4, Betaine and nucleic acid test strips. The primer has the advantages that of being easy and convenient to use, rapid, high in specificity and sensitivity, objective and visual in result judgment, low in cost, convenient to use and quite safe to people and environment. The cyprinid herpes virus type 2 CPA detection primer not only can be used in a special laboratory, but also can be applied in wild on-site rapid detection, and the cyprinid herpes virus type 2 in a sample can be accurately detected within 1.5 h through only one metal bath or water bath pot.

The invention belongs to the technical field of nucleic acid test sampling, and provides a flexible operating head, a rigid base and a facility for nucleic acid test sampling. On the basis of the principle that a cavity formed inside a flexible material can deform in the air inflation and deflation and / or pumping process, the flexible operating head with an air cavity structure is obtained and used for allowing a throat swab to be installed and controlling the throat swab to conduct sampling operation, and therefore a medical worker does not need to hold the throat swab to conduct operation byhimself in the sampling process. The flexible operating head can cooperate with the rigid base and a mechanical arm to be used, and therefore the facility which can allow the medical worker to conduct long-distance sampling operation is obtained, the medical worker is prevented from touching a patient in a short-distance mode, and the infection risk is reduced. In addition, the replacement of thedisposable flexible operating head can be automatically accomplished on the basis of the magnetic attraction connection principle, the process is safe and effective, and the cost is low.

The invention relates to a method for detecting clostridium perfringens based on a nucleic acid chromatography biosensing technology. According to a virulence gene cpa of clostridium perfringens, loop-mediated isothermal amplification primers (SEQ ID NO:1-4) are designed, and through the combination with a nano-enzyme nucleic acid test strip, the clostridium perfringens detection method based on an LAMP nano-enzyme sensor is established. The method for detecting clostridium perfringens based on the nucleic acid chromatography biosensing technology can be successfully used for distinguishing live bacterial cells from dead bacterial cells, and the lower limit of detection on clostridium perfringens can reach 10 CFU / mL.

The invention provides a nucleic acid test card and an application method thereof. The nucleic acid test card comprises a sealing film, a card body, a reagent hole, a nucleic acid amplification reagent, reaction holes, nucleic acid collection test paper, a nucleic acid purifying reagent for matching use, a washing buffer and an organic phase for preventing a solution from being volatized. The application method comprises the following steps: opening the sealing film, adding a pathogene sample into the reaction holes packaged with the nucleic acid collection test paper, automatically washing by the use of the nucleic acid purifying reagent and the washing buffer for several times, respectively adding the nucleic acid amplification reagent in the reagent hole into each corresponding reaction hole to carry out a nucleic acid amplification reaction under the protection of the organic phase for preventing a solution from being volatized, and carrying out qualitative and quantitative analysis on the reaction result through an optical inspection instrument for matching use. By the adoption of the test card and through a nucleic acid amplification reaction, simultaneous testing of a single sample or various pathogens or target genes in various samples is realized. Time consumption of the test is low, test accuracy and sensitivity are high. The test card is applicable in the fields of experiment research, clinical test and food safety testing, etc.