10 results about "NASBA" patented technology

The invention discloses a method for rapidly detecting mycobacteria. The method comprises the following steps: using a primer pair for detecting the Mycobacterium tuberculosis, wherein the primer pair comprises an upstream primer and a downstream primer; the upstream primer comprises a nucleotide sequence as shown in SEQ ID NO: 1 or 2, and the downstream primer comprises a nucleotide sequence as shown in any one of SEQ ID NO: 3-6. According to the present invention, the RNA of the MTB viable bacteria in the biological sample can be rapidly and accurately detected with high sensitivity, the detection limit is reduced by 1000 times compared with the existing NASBA kit, the template requirement is reduced, and the better choice is provided for the instant detection of MTB.

A nucleic acid extraction composition, reagents and kits containing the same and uses thereof. Provided is a nucleic acid extraction and purification reagent free of volatile organic solvents, which prevents the damage of volatile organic solvents to people and greatly improves the timeliness of nucleic acid extraction and purification, making the operation extremely simple, and the nucleic acid can be obtained within 10 minutes. The obtained nucleic acid may be used for biological reactions such as PCR, NASBA, LAMP and RPA. Moreover, the reagents of the present disclosure may be used to extract nucleic acids of cells, bacteria, fungi, DNA viruses and RNA viruses from various samples such as blood, throat swab preserving fluid, saliva, urine, sputum, excrement and the like, and very suitable for clinical and scientific research uses.

Provided is a method for rapidly detecting Mycobacterium tuberculosis. The method comprises using a primer pair for detecting Mycobacterium tuberculosis, which primer pair comprising an upstream primer and a downstream primer; wherein the upstream primer comprises a nucleotide sequence as shown in SEQ ID NO: 1 or 2, and the downstream primer comprises a nucleotide sequence as shown in any one of SEQ ID NOs: 3-6. The method can rapidly and accurately detect the RNA of MTB viable bacteria in a biological sample with high sensitivity, reduces the detection limit by up to 1000 times compared with existing NASBA kits, reduces the requirements for templates, and provides a better option for the real-time detection of MTB.

The invention discloses a universal probe for multiple nucleic acid detection, a probe combination and application of the universal probe and the probe combination, aiming at the limitation that the existing nucleic acid detection technology depends on expensive equipment, is complicated to operate and is difficult to realize multiple detection. A bridging detection system is formed by innovatively designing a general probe, a specific probe series A (two ends of which are fixed in sequence and are used for connecting the general probe and a quality control line) and a specific probe series B (one end of which is fixed on a detection line); the test strip structurally comprises a sample pad, a colloidal gold pad (for fixing a universal probe), an NC (nitrocellulose) membrane (containing a plurality of detection lines and quality control lines) and absorbent paper. The method is suitable for single-stranded nucleic acid (such as NASBA amplification products) detection, has high sensitivity, high specificity (no false positive), rapidness (completion within 10 minutes) and multiple detection capability, does not need complex instruments, is low in cost, and is easy to popularize in a basic level.

The invention discloses a hand-foot-and-mouth disease virus isothermal amplification detection method, and relates to the technical field of virus detection. The hand-foot-and-mouth disease virus isothermal amplification detection method comprises three steps, by adopting the LAMP technology, the specificity is high, the sensitivity is higher than that of a PCR detection method, the method is an improvement and supplement of an existing nucleic acid molecule detection technical means, the method has the advantages of being simple, rapid, high in sensitivity and high in specificity, primers are designed according to the specific sequence of the hand-foot-and-mouth virus, and the detection efficiency is improved. Compared with a traditional culture method, the method has the advantages that the specificity is higher, the sensitivity is high, the operation is simple and convenient, the price is low, the end point detection of an NASBA amplification result is realized by combining a handheld fluorescence detector while the real-time fluorescence detection is met, and the detection sensitivity reaches the fluorescent quantitative PCR level.

The present invention relates to a method for detecting a target nucleic acid in a sample using an optimized SMART method that involves the use of at least one fluorescence-generating aptamer for the direct detection of RNA transcripts. Preferably, the method is used after an amplification step, for example, after a NASBA assay.

The present invention belongs to the technical field of genetic engineering, in particular to a method for detecting classical swine fever virus based on a G4-ThT biosensor and NASBA and a kit thereof. The specific method includes using the viral RNA extracted from a classical swine fever virus sample as the amplification target, performing isothermal amplification using an isothermal nucleic acid amplification technique, then adding a buffer solution containing Tris-HCl and KCl, and further adding thioflavin T to promote the folding of G-quadruplexes on the amplification products to form a complex, detecting fluorescence under the excitation of a light source with a specific wavelength, and making a judgment according to the intensity thereof. The present invention provides a probe that does not require special biotin labeling or thiol modification treatment, a detection method with significantly reduced detection costs, and a kit.

The invention relates to the technical field of virus detection, and provides a swine influenza virus detection reagent, a swine influenza virus detection kit and a swine influenza virus detection method.The swine influenza virus detection reagent and the swine influenza virus detection kit comprise a pair of NASBA amplification primers, an NASBA amplification system and NASBA enzyme reaction liquid; according to the method, an NASBA isothermal amplification system and a Mango detection system are coupled in a tubular detection mode, detection can be carried out without adding a fluorescent group labeled probe, the detection cost is low, the sensitivity and accuracy are high, the specificity is good, the detection operation is simple, rapid and high in efficiency, and the method is suitable for rapid detection of the swine influenza virus.

The application discloses a gene and drug resistance mutation detection method based on a double-site nucleic acid circuit probe and belongs to the technical field of molecular biology. In order to solve the technical problems of false positive detection results, complex operation, high requirements for detection personnel and high cost in the prior art gene and drug resistance mutation detection method, the application provides a gene and drug resistance mutation detection method based on a double-site nucleic acid circuit probe. The method is based on isothermal amplification technology combined with one or more nucleic acid circuits. The isothermal amplification technology includes but is not limited to NASBA, SDA, LAMP, HDA and RPA. The nucleic acid circuit includes but is not limited to a cascade strand displacement nucleic acid circuit and a one-step strand displacement nucleic acid circuit. The detection method provided by the application can simultaneously detect multiple drug resistance mutation sites through a single tube and a single reaction, realizes pathogen detection and pathogen drug resistance mutation detection on unknown samples, has high detection accuracy, is simple in steps and low in cost.

The present invention discloses a detection technology for quantitative live bacterial RNA single nucleotide polymorphism, which belongs to the field of nucleic acid detection. This method integrates nucleic acid sequence-dependent amplification technology (NASBA), allowing rapid and specific generation of target RNA amplification, and amplification hindrance mutation system (ARMS), which introduces mismatches at the 3' end base of the forward primer to enhance the specificity of identifying single nucleotides, thereby highly selectively identifying RNA mutations, and can sensitively identify single nucleotide polymorphisms with abundance as low as 0.5%, and detect live Salmonella as low as 80 CFU. This technology allows RNA to be amplified under isothermal conditions and single nucleotide polymorphisms on RNA to be detected in a single tube, and can be used to evaluate the presence and vitality of drug-resistant bacteria under the action of disinfectants, and has good application prospects.