37 results about "NASBA" patented technology

The invention discloses an NASBA-ELISA (nucleic acid sequence-based amplification-enzyme-linked immuno sorbent assay) detection primer, a probe and a kit for type II grass carp reovirus. Nucleotide sequences of the primer and the probe are as shown in SEQ ID NO.1-4. The kit contains the primer, the probe, an NASBA amplification buffer solution, an enzyme mixed liquor, an amplification product detection reagent, a negative control and a positive control. The kit for the type II grass carp reovirus has the advantages that a detection time period is short, and detection efficiency is high; virus detection specificity is high, and accuracy is high; virus quantitative analysis can be realized while virus qualitative analysis is realized; detection sensitivity is higher than that of common PCR (polymerase chain reaction); operation is easy, and popularization is easy; and repeatability of experimental results is good.

The invention relates to an enteropathogen rapid detection technology based on real-time nucleic acid sequence-based amplification (NASBA). Specifically, the invention provides an Astrovirus real-time isothermal amplification detection kit, and a pair of primers and a molecular beacon probe thereof. The kit includes: a 2*real-time NASBA reaction solution containing the primers and the probe, a 5*enzyme mixed solution and a positive control template, a negative control and a blank control. The sequences of the primers and the probe are the sequences numbered as SEQIDNO:1-3, and the primers and the probe can specifically amplify and detect an Astrovirus ORF2 gene fragment. The kit provided in the invention has the characteristics of fastness, high efficiency, sensitivity and specificity, and real-time detection analysis, etc., and can be used in the fields of conventional detection and disease control and prevention in clinical practice and ports.

The invention relates to the field of virus detection, and particularly relates to a virus nucleic acid extraction or preservation reagent, a primer probe combination, a virus amplification reagent, akit and application thereof. A virus detection device provides a simple and feasible virus nucleic acid extraction method, the whole process is about 5-15 minutes, and purified nucleic acid is recovered and can be used for detecting virus nucleic acid. The method comprises PCR, NASBA, LAMP, RPA and the like. Compared with a traditional virus extraction method, the method is high in virus nucleicacid recovery rate, short in time, convenient to operate and easy to clinically popularize. The invention relates to an isothermal amplification primer, a probe combination sequence and a reaction buffer solution for simultaneously detecting novel coronavirus COVID-19N and ORF genes and human-derived reference genes through a single tube. The system is good in specificity, high in sensitivity (50cp/mL) and high in specificity, only needs 20min of detection time, and can report positive within about 10 min at the soonest.

The invention discloses a nucleic-acid sequence-based amplification (NASBA) method for detecting a swine influenza virus (SIV). The NASBA method mainly comprises the following steps of: extracting the ribonucleic acid (RNA) from the SIV, preparing an NASBA system, preparing an NASBA program, carrying out identification on an NASBA product and carrying out PCR (Polymerase Chain Reaction) amplification. According to the NASBA method, the NASBA rapid detection method is established by taking the NP gene of the SIV as a target gene and is used for the diagnosis of the SIV. The method has higher specificity and sensitivity so as to detect a virus solution with the dilution factor of 10<-5>, so that the method can be used for the rapid detection of clinically-suspected SIV samples, meanwhile, the technical level of China in the diagnosis, epidemic surveillance, inspection and quarantine of swine influenza can be improved so as to ensure the healthful and rapid development of swine industry.

The invention discloses a swine fever virus detection method based on G-quadruplex fluorescence characteristic and a kit, and belongs to the technical field of gene engineering. The method comprises: utilizing a nucleic acid sequence-based amplification (NASBA) technology to amplify target RNA to obtain a single-chain RNA product; adding an upstream probe, a downstream probe and the RNA amplification product into a G- quadruplex fluorescence detection buffer, performing denaturation and annealing, then adding protoporphyrin, and utilizing a microplate reader or fluorophotometer to detect the fluorescence intensity in the reaction system, so as to determine whether the sample possesses a swine fever virus. The method is fast, precise and stable in swine fever virus detection speed, good in reappearance, simple in detection steps and low in cost.

The invention discloses a Macrobrachium rosenbergii Nodavirus NASBA-LFD (nuclear acid sequence-based amplification-lateral flow dipstick) detection method and a detection kit thereof. The detection method includes: according to a Macrobrachium rosenbergii Nodavirus (MrNV) sequence published by GenBank, screening a conserved region, and designing the primers and probe needed by an MrNV NASBA-LFD reaction system; employing a tissue RNA extraction reagent prepared by the inventor to extract the RNA of a to-be-detected sample, then utilizing the MrNV NASBA reaction system established by the invention to perform detection, and at the end of the reaction, determining the result according to nucleic acid rapid test paper. Compared with common detection technologies of the virus, the method and the kit provided in the invention have the characteristics of simplicity and rapidity, good specificity and high sensitivity, can be used for field service by technical personnel and farmers at the production front line, and are in favor of diagnosis prevention and pathogen blocking of Macrobrachium rosenbergii white tail disease.

The invention discloses an NASBA detection primer and an NASBA detection method for identifying the 1/2c serotype of listeria monocytogenes, belonging to the field of biotechnology. A forward primer sequence is 5'-GAAATACATTTTGGTCTAG-3', a reverse primer sequence is 5'-aattctaatacgactcactatagggATTACTGGGAATAACTTGA-3', a sequence of a molecular beacon probe for detecting the 1/2c serotype of listeria monocytogenes is 5'-JOE-CGATCGAGGAAATGCTTATGCAGATATTACGATCG-DABCYL-3'. The detection method comprises the following steps: extracting RNA of listeria monocytogenes, carrying out a real-time NASBA reaction, and carrying out qualitative analysis. The constructed primer and molecular beacon have high specificity and sensitivity for the identification of the 1/2c serotype strain of the listeria monocytogenes, for the detection method, the operation steps are simple, the detection time is short, real-time qualitative analysis can be carried out, and meanwhile, living bacteria and dead bacteria can be effectively distinguished.

A method for detecting the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in a biological sample includes the steps of lysing the biological sample to form a lysate and generating an amplified lysate by performing a nucleic acid sequence-based (NASBA) amplification for a target nucleic acid sequence in the lysate in the presence of: a forward primer having the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 9, SEQ ID NO: 13, and SEQ ID NO: 17; a reverse primer having the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 10, SEQ ID NO: 14, and SEQ ID NO: 18; and a molecular beacon having the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 7, SEQ ID NO: 11, SEQ ID NO: 15, and SEQ ID NO: 19 and a fluorophore. The amplified lysate is exposed to an excitation source. A fluorescence of the fluorophore is detected in the amplified lysate exposed to the excitation source The SARS-CoV-2 is determined to be present in the biological sample in response to detecting the fluorescence of the fluorophore.

The invention discloses a method for detecting REV by using NASBA, and a reagent set used therein. The reagent set is composed of a primer pair including a primer F and a primer R, and a probe set. A target sequence of the primer pair contains a specific DNA fragment as shown in SEQ ID NO: 9. The probe group consists of an upstream probe as shown in SEQ ID NO: 3 and a downstream probe as shown in SEQ ID NO: 4. The reagent set provided by the invention is used for detecting the REV, the specificity is good (the REV is accurately identified from ALV, MDV, CIAV and REV), the sensitivity is high (the sensitivity is 2 * 10 copy number/reaction system), the reagent set is suitable for detecting the REV in a sample with low virus content, and the detection method is simple, convenient, rapid and accurate (the detection is completed within 1 hour). The method and the reagent set have great popularization value.

The invention discloses a 2019-nCoV nucleic acid isothermal amplification detection kit based on SYBR Green I and a detection method. The kit comprises an ORF1ab gene primer sequence and an N gene primer sequence of a 2019-nCoV virus, an SYBR Green I dye, an NASBA amplification buffer solution and an NASBA enzyme mixed solution. The detection method using the kit comprises the following steps: firstly, preparing the NASBA amplification buffer solution, preparing the NASBA enzyme mixed solution, preparing a primer mixed solution and preparing an RNA sample; and carrying out mixed reaction on all the solutions, carrying out incubating, and finally completing detection by using a fluorescence detector. The kit and the method show high-specificity and high-sensitivity detection performance, and a large amount of experimental time is saved.

The invention discloses a NASBA primer, a reagent kit, and a method for detecting apple scar skin viroid; the sequences of the upstream primer and the downstream primer are respectively SEQ ID NO: 1-2; the NASBA augmentation reagent kit comprises the constituents as follows: (1) NASBA augmentation reaction liquid A; (2) NASBA augmentation reaction liquid B; the detection method is as follows: 1) extracting sample RNA; 2) augmenting NASBA of ASSVd; 3) detecting with electrophoresis. The NASBA primer, the reagent kit, and the method for detecting apple scar skin viroid are applicable to quickly detecting and confirming apple scar skin viroid, can be widely applied for epidemic surveillance in agricultural production and environment as well as monitoring and detecting the viroid in import and export trade; the operation is very simple; the amount of the required sample is small; and the quality requirements to template RNA is lower.

A system for detecting the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in a biological sample includes a sampling device, a lysing chamber, a NASBA fluidic network, and an analytical instrument. The sampling device is configured to contain a sample containing a pathogen target sequence for SARS-CoV-2. The lysing chamber is configured to be in fluid communication with the sampling device to receive the sample. The is lysing chamber is configured to lyse the sample into a lysate. The NASBA fluidic network is configured to be in fluid communication with the lysing chamber to receive the lysate. The NASBA fluidic network includes an enzyme, a forward primer, and a reverse primer for amplifying a predetermined genetic sequence in the pathogen target sequence contained within the lysate. The forward primer has the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 9, SEQ ID NO: 13, and SEQ ID NO: 17. The reverse primer has the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 10, SEQ ID NO: 14, and SEQ ID NO: 18. A molecular beacon is configured to attach to the pathogen target sequence. The beacon has the oligonucleotide sequence selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 7, SEQ ID NO: 11, SEQ ID NO: 15, and SEQ ID NO: 19 and a fluorophore. The analytical instrument is configured to excite the beacon when the molecular beacon is attached to the pathogen target sequence to signal a presence of the pathogen target sequence.