16 results about "RNA amplification" patented technology

The invention discloses a digital micro-droplet RNA amplification detection system for prostatic cancer and application of PCA3 combined with an ERG gene in the system, and belongs to the technical field of tumor diagnosis. The digital micro-droplet RNA amplification detection system for prostatic cancer comprises reagents for respectively and specifically detecting the following genes based on a real-time fluorescent nucleic acid isothermal amplification method: PCA3, ERG and SPDEF, and a digital PCR system. Compared with the existing kit or other products for detecting the prostate cancer, the digital micro-droplet RNA amplification detection system provided by the invention has the advantages that the detection omission ratio of the prostate cancer is greatly reduced, and the detection sensitivity and the negative predictive value of the prostate cancer are improved.

Synthetic endogenous ribonucleic acid virus satellite RNA molecules and satellite particles containing the same are disclosed. A synthetic endogenous ribonucleic acid virus satellite RNA molecule containing an endogenous heterologous TNA virus (HRV) amplicon is also disclosed. Also disclosed are methods of using endogenous ribonucleic acid virus satellite RNA molecules and satellite particles containing the same to alter plant phenotypes, improve stress resistance in plants, and improve pest and pathogen resistance in plants.

Disclosed herein is a recombinant vector for expressing a replicase comprising a novel polydeoxyribonucleotide. The thus-expressed replicase can be used in in vitro RNA amplification via an RNA-dependent RNA cycling reaction (RCR). Also disclosed herein is a method for producing an amplified RNA product in an RNA cycling reaction (RCR) via use of the present replicase encoded by the present polydeoxyribonucleotide.

This disclosure provides methods of measuring trans-amplifying RNA and self- amplifying RNA amplification in vivo. The taRNA and the saRNA are barcoded, administered to a subject, and barcode amplification in a specific cell type is measured.

Disclosed in the present invention are a digital droplet RNA amplification detection system for prostate cancer and a method for reducing the false negative rate of prostate cancer. The provided digital droplet RNA amplification detection system for prostate cancer comprises reagents for respectively and specifically detecting the following genes on the basis of a real-time fluorescent nucleic acid constant-temperature amplification method: PCA3, ERG and SPDEF, and a digital PCR system.

A method for sample preparation and / or DNA amplification from urine, saliva and / or mouthwash samples is disclosed. The method involves providing a sample preparation system comprising a filter and providing a urine, saliva and / or mouthwash sample, then filtering at least a portion of the sample through the filter, thereby retaining bacteria, fungi, viruses protists and / or cancerous or precancerous cells in the filter. Subsequently a lysis reagent is applied to the filter. Finally, the DNA found in the lysate is amplified.

This invention covers processes that amplify multiple DNA and RNA targets, solving the “multiplexed PCR problem”, as well as supporting isothermal amplification and delivering other capabilities in multiplexed molecular analysis. It uses inventive DNA analogs.

Synthetic split viral satellite RNA molecules and satellite particles containing the same are disclosed. Also disclosed are synthetic split viral satellite RNA molecules containing an internal heterologous TNA virus (HRV) amplicon. Also disclosed are methods of using the split viral satellite RNA molecules and satellite particles containing the same to alter plant phenotypes, improve stress resistance in plants, and improve pest and pathogen resistance in plants.

The invention relates to the technical field of biology, in particular to a primer group and a method for quantitatively amplifying and detecting HBV RNA in serum or plasma. The invention provides a blocking primer, a primer group, a reagent and a kit for RNA amplification of hepatitis B virus and application of the blocking primer, the primer group, the reagent and the kit. The blocking primer for hepatitis B virus RNA amplification is obtained through screening, the optimal primer group for hepatitis B virus RNA amplification is obtained through further optimization, the primer group is used for hepatitis B virus RNA amplification, it can be guaranteed that specific amplification is selectively carried out only on HBV RNA in HBV DNA and RNA genome mixed nucleic acid, time and labor are saved, and the detection efficiency is improved. The method is good in repeatability, good in accuracy, high in sensitivity and low in detection limit, can be matched with an automatic device for use, and has a wide application prospect.

The application discloses a recombinant Bst DNA polymerase with high DNA polymerase activity and reverse transcriptase activity, and preparation and application thereof. Sulfolobus tokodaii The application carries out sequence rearrangement on a Headpiece domain truncated body Hp47 derived from chicken villin and a Sto7d domain derived from a thermophilic archaea, and fuses the amino terminal end of the Bst DNA polymerase large fragment to construct a novel Bst DNA polymerase rearrangement mutant HpSt-G23. The recombinant polymerase HpSt-G23 obtained by the application has high polymerase activity and reverse transcriptase activity, and can resist inhibitors such as urea and sodium chloride. Tth The HpSt-G23 can be used in a RecA detection system, and only two primers are needed to complete DNA or RNA amplification in the same system, and the detection sensitivity of the RNA template is greatly improved, so that the HpSt-G23 has a good development prospect in the field of nucleic acid detection.

The invention discloses a digital micro-droplet RNA amplification detection system for detecting Alzheimer's disease as well as a special primer and a probe thereof, and belongs to the technical field of senile disease diagnosis. The digital micro-droplet RNA amplification detection system for the Alzheimer's disease comprises reagents for respectively and specifically detecting the following genes based on a real-time fluorescent nucleic acid isothermal amplification method: TRPC6 and Gapdh, and a digital PCR (Polymerase Chain Reaction) system. The digital micro-droplet RNA amplification detection system provided by the invention can be used for detecting the Alzheimer's disease by taking the blood of a patient as a sample to be detected, and has extremely high sensitivity and a negative predicted value, so that the diagnosis accuracy of the Alzheimer's disease is greatly improved.

The present invention provides a method for single-cell transcriptome sequencing. Specifically, the method of the present invention adopts the "one-tube method" to complete the sample lysis, reverse transcription, PCR amplification and product purification steps in the same reaction vessel, thereby improving the sensitivity of amplification of low-abundance RNA in the cell transcriptome and RNA in special structures, reducing the loss of transcriptome information, and improving the efficiency of single-cell transcriptome amplification and sequencing. In addition, the method of the present invention is applicable to transcriptome sequencing of fixed single-cell samples and live fresh single-cell samples. The present invention also provides a kit for single-cell transcriptome sequencing and a single-cell transcriptome sequencing device, which realizes more convenient, rapid, sensitive and complete sequencing of single-cell transcriptomes.

The invention belongs to the technical field of molecular biological detection, and discloses a reaction system, a kit and a detection method applied to RNA amplification of a complex whole blood sample. The reaction system comprises a buffer component and an enzyme protection component; the buffer component consists of Tris and Bicine and is used for cooperatively maintaining the stability of the pH value of a reaction system in the RNA amplification process of a complex whole blood sample; and the enzyme protection component is composed of AEBSF and dextran sulfate. Through collaborative innovation of a double-buffer system and an active enzyme protective agent, the integrated challenges that the pH is easy to drift, the template compatibility is poor and the enzyme activity is easy to damage when a complex whole blood sample is faced in the prior art are effectively overcome, and a key technical support is provided for rapid, stable and reliable bedside molecular diagnosis.

The invention discloses a recombinant Bst DNA polymerase with high DNA polymerase activity and reverse transcriptase activity as well as preparation and application of the recombinant Bst DNA polymerase. According to the present invention, a Headpiece structural domain truncated body Hp47 derived from chicken villus protein and a Sto7d structural domain derived from thermophilic archaea Sulfolobus tokidaii are subjected to sequence rearrangement, and are fused to the amino terminal of a Bst DNA polymerase large fragment so as to construct the novel Bst DNA polymerase rearrangement mutant HpSt-G23; the recombinant polymerase HpSt-G23 obtained by the invention has both high polymerase activity and reverse transcriptase activity, and can tolerate inhibitors such as urea, sodium chloride and the like. HpSt-G23 and TthRecA can construct a detection system, DNA or RNA amplification can be completed in the same system only by using two primers, the detection sensitivity of an RNA template is greatly improved, and the HpSt-G23 has a good development prospect in the field of nucleic acid detection.

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.

Provided are a digital microdroplet RNA amplification detection system and method for reducing the missed detection rate of prostate cancer, which belong to the technical field of tumor diagnosis. The provided digital microdroplet RNA amplification detection system for reducing the missed detection rate of prostate cancer comprises: reagents based on a real-time fluorescent nucleic acid isothermal amplification method for specifically detecting the following genes, respectively: PCA3, ERG, and SPDEF; and a digital PCR system. Compared with existing kits or other products for prostate cancer detection, the provided digital microdroplet RNA amplification detection system can significantly reduce the missed detection rate of prostate cancer and improve the detection sensitivity and negative predictive value for prostate cancer.