44 results about "RNase P" patented technology

A viral vector production system is provided which system comprises: (i) a viral genome comprising at least one first nucleotide sequence encoding a gene product capable of binding to and effecting the cleavage, directly or indirectly, of a second nucleotide sequence, or transcription product thereof, encoding a viral polypeptide required for the assembly of viral particles, (ii) a third nucleotide sequence encoding said viral polypeptide required for the assembly of the viral genome into viral particles, which third nucleotide sequence has a different nucleotide sequence to the second nucleotide sequence such that said third nucleotide sequence, or transcription product thereof, is resistant to cleavage directed by said gene product; wherein at least one of the gene products is an external guide sequence capable of binding to and effecting the cleavage by RNase P of the second nucleotide sequence. The viral vector production system may be used to produce viral particles for use in treating or preventing viral infection.

The invention belongs to the technical field of virus detection, and particularly relates to a primer, probe, test kit and method for real-time fluorescent quantitative PCR detection of novel coronavirus 2019-nCoV. A single-tube double-fluorescence channel is adopted to simultaneously detect the existence of the novel coronavirus 2019-nCoV and reference gene Rnase P, and the existence of the novelcoronavirus 2019-nCoV RNA in specimens such as alveolar lavage fluid, nasopharyngeal swab, whole blood, serum, feces and tissues can be detected. The method is short in detection time period, and issuitable for clinical and bedside rapid detection of diagnosis; the virus detection specificity is high, and the accuracy is high; virus qualitative analysis and quantitative analysis are carried out,and the quantitative linear range is good; the detection sensitivity is high; the experimental result is good in repeatability and high in precision; and the reference gene is added into a detectionsystem, so that the quality of the whole process of extracting and amplifying the sample can be monitored through the detection result of the reference gene.

The invention discloses novel coronavirus 2019-nCoV fluorescent RPA detection primers, a probe, a kit and a method. The kit comprises novel coronavirus 2019-nCoV specific primers shown in SEQ ID NO. 1-2 and a probe shown in SEQ ID NO. 3. According to the invention, the existence of the novel coronavirus 2019-nCoV ORF1ab gene and the internal reference gene RNase P is simultaneously detected by adopting a single-tube double-fluorescence channel, and the existence of the novel coronavirus 2019-nCoV RNA in pulmonary alveolar lavage fluid, nasal swab, pharyngeal swab, sputum and other samples canbe detected. The method is short in detection time and suitable for clinical and bedside virus nucleic acid rapid screening, the kit has extremely high sensitivity and specificity, an internal reference gene is added into the detection system, and quality monitoring is performed on the extraction and amplification process of the sample according to the detection result of the internal reference gene.

The invention discloses a high-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit and a use method. The high-sensitivity novel coronavirus 2019-nCoV nucleic acid detection kit comprises an ORF1ab gene primer pair, an N gene primer pair, an endogenous reference gene primer pair and a probe set. Novel coronavirus (2019-nCoV) ORF1ab and N genes are used as target areas, specific primers and probes are designed; by detecting the change of fluorescence signals, the novel coronavirus 2019-nCoV nucleic acid in a sample is subjected to qualitative detection; the 5' ends of the ORF1ab gene probe and the N gene probe are modified by a fluorophore FAM to form a monochromatic double-target detection system, and a large-volume system higher than the conventional system is used for amplification, so that when nucleic acid detection is carried out on the novel coronavirus 2019-nCoV, the detection sensitivity is high and can reach 50 copies/mL; and moreover, the nucleic acid extraction and detection process is monitored by taking a ribonuclease P (RNase P, RNP) gene widely existing in normal human epithelial cells as an internal target gene, and false negative and false positive are avoided, so that the nucleic acid detection kit is wide in market popularization prospect.

The invention discloses a gastric juice multiple real-time polymerase chain reaction (PCR) detection-based helicobacter pylori (HP) individualized treatment auxiliary diagnosis method. According to the method, specific genes of HP, HP clarithromycin drug-resistant mutation sites A2143G and A2142G, and polymorphism of CYP2C19 gene sites such as CYP2C19*2(G681A) and CYP2C19*3(G636A) of corresponding patients can be simultaneously detected from 10-150mu l of gastric juice samples, and human epithelial cell ribonuclease P(Rnase P) is taken as a reference gene. The detection method is rapid, accurate and small in sample demand amount, whether HP infection exists can be detected from the 10-150mu l of gastric juice samples in a single test, the HP clarithromycin drug-resistant conditions and genotypes of CYP2C19 of the patients can be reported, and medication guide is provided for individualized eradication therapy of HP infection.

The invention relates to the field of nucleic acid detection, and particularly discloses a nucleic acid composition, a kit and a method for simultaneously detecting multiple mutant strains of SARS-CoV-2. The SARS-CoV-2 comprises a wild type and Alpha mutant strain, a Beta mutant strain, a Gamma mutant strain, an Eta mutant strain, a Delta mutant strain, a lota mutant strain, a Kappa mutant strain and a Lambda mutant strain, and the design sites of a primer probe composition comprise N gene, RNase P gene and S gene N501Y, E484K, K417T, L452R, E484Q, F490S, T76I, L452Q and 246-253del mutant types. The nucleic acid composition, the kit and the method disclosed by the invention can be used for simultaneously and rapidly detecting the SARS-CoV-2 and the main epidemic mutant strain thereof, and have the advantages of high sensitivity and good specificity.

The invention discloses a multi-fluorescent PCR rapid detection kit for novel coronavirus, influenza A virus and influenza B virus. The kit comprises freeze-dried solid RT-PCR Mix, liquid redissolution Buffer, freeze-dried solid positive control and freeze-dried solid negative control, wherein the freeze-dried solid RT-PCR Mix contains a primer group corresponding to primer sequences of a SARS-CoV-2 specific gene ORF1ab, an influenza A M gene, an influenza B M gene and a human reference gene RNAse P. The kit combines a multi-fluorescent quantitative PCR technology and a freeze-drying process, utilizes three pairs of special primers and human reference genes to amplify specific sequences of three pathogens in vitro, and performs real-time detection in combination with a fluorescent probe. The detection method is simple and convenient to operate, has low requirements for the operation level of detection personnel, and can detect three common respiratory pathogens at a time, the detection time and the detection cost are greatly saved, rapid screening of large-batch samples is realized, the whole detection process only takes 40 minutes to 1 hours, and results are accurate and reliable.

The invention discloses an LTL (leukocyte telomere length) detecting method based on a qPCR (quantitative polymerase chain reaction) method. The method comprises steps as follows: 1) extracting genomic DNA of different tissue, diluting DNA samples to 5 ng/ul, taking 5 ul of each tissue sample, and mixing the tissue samples to prepare a whole genomic DNA template; 2) performing primary real-time qPCR detection and detecting a telomere primer of a telomere repetitive sequence; 3) performing secondary real-time qPCR detection and transcribing a single-copy reference RNase P gene of RNase P enzyme; 4) calculating the relative telomere length T/S. The LTL detecting method based on the qPCR method has the benefits as follows: the requirement for the samples and genomic DNA concentration is not high, the experiment operation process is simple and feasible, the result credibility is high, and the method is adaptable to scientific research and clinical popularization and application and plays an important role in preventing aging, cancer and diseases.

The invention discloses an influenza A and B virus nucleic acid detection kit. The influenza A and B virus nucleic acid detection kit comprises a nucleic acid amplification reaction solution, an enzyme mixed solution, an influenza A/B virus mixed reaction solution, a positive control solution, a weakly positive control solution and a negative control solution, wherein the influenza A/B virus mixedreaction solution comprises an influenza A virus upstream primer, an influenza A virus downstream primer, an influenza A virus probe, an influenza B virus upstream primer, an influenza B virus downstream primer, an influenza B virus probe, an internal reference RNase P upstream primer, an internal reference RNase P downstream primer, an internal reference RNase P probe and DEPC water, and the internal reference means the detection of the internal reference of the product. The internal reference of the embodiment participates in the process from nucleic acid extraction to RT-PCR, and can be used for monitoring the collection, storage and transportation of samples and the nucleic acid extraction process and amplification process, so that false negative interpretation results are avoided.

The invention discloses an EGS of 12nt, which can conduct RNase P ribozyme and comprises EGS which is based on RNA and has the nucleotide sequence to be shown in SEQ ID NO: 1, and EGS that is based on DNA and has the nucleotide sequence to be shown in SEQ ID NO: 2. The EGS can effectively inhibit the expression of HCMV UL49 gene. The EGS with the characteristic of DNA has the silent gene efficiency of 99%, and the EGS with the characteristic of RNA has the silent gene efficiency of 98%. Compared with the existing EGS which can cause the UL49 gene to be silent, the EGS of the invention has the sequence which only contains 12 basic groups, thus not only having the shortest nucleic acid molecules, but also effectively and specially leading the expression of the UL49 gene to be silent. The EGS of the invention is used for researching and developing the anti-HCMV drug, and can greatly reduce the cost when the drug is researched and applied.

The invention provides a freeze-dried type multiple fluorescent RT-PCR reagent and a method for detecting and distinguishing influenza A virus, influenza B virus and coronavirus SARS-CoV-2. Primers and probes of the fluorescent RT-PCR reagent for detecting the influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 provided by the invention are obtained by designing and screening based on the latest gene sequence information of the influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 in a public gene sequence database, and the reagent has good inclusivity, specificity and reaction performance. According to the reagent, a human RNase P nucleic acid fragment is used as a detection internal reference. In addition, the fluorescent RT-PCR detection reagent forthe influenza A virus, the influenza B virus and the coronavirus SARS-CoV-2 is prepared into a freeze-dried form capable of being stored at normal temperature through freeze-drying technology, thus avoiding the risk that the conventional liquid fluorescent PCR detection reagent is invalid due to the change of storage temperature or repeated freezing and thawing, and the transportation and management cost is reduced.

Synergistic pharmaceutical compositions including an RNase P inhibitor and a tRNA synthetase inhibitor are provided, as well as methods for their use in treating infections. Also provided herein are methods of using the compositions to inhibit a bacterial tRNA synthetase in a cell and to decolonize bacteria on a surface.

The invention discloses a mycoplasma genitalium nucleic acid real-time fluorescence PCR detection primer, a probe and a kit. The kit comprises an amplification reaction solution, a mycoplasma genitalium nucleic acid detection solution, a positive reference substance and a negative reference substance, wherein the mycoplasma genitalium nucleic acid detection liquid comprises specific primers and probes aiming at mycoplasma genitalium and a human RNase P gene, the sequences of the primers aiming at the mycoplasma genitalium are as shown in SEQ ID NO.1 and SEQ ID NO.2, and the sequence of the probe aiming at the mycoplasma genitalium is as shown in SEQ ID NO.3. According to the primer, the probe and the kit for real-time fluorescence PCR detection of the mycoplasma genitalium, provided by the invention, the mycoplasma genitalium can be rapidly, accurately and sensitively detected, the repeatability of an experimental result is good, the precision is high, the detection time period is short, and the detection can be completed within 70 minutes as short as possible. Therefore, the detection time is greatly saved, and the clinical diagnosis efficiency can be accelerated.

The invention discloses a novel coronavirus 2019-nCoV real-time fluorescent PCR detection primer and probe, kit and method. Two loci of nucleic acid after 2019-nCoV replication are selected for detection; the two loci include a 2019-nCoV specific locus with high specificity, and the locus has no cross reaction with other common pathogens of which infection site are the same as the locus or have similar infection symptoms and the total nucleic acid of human leukocytes. A single tube with three fluorescence channels are adopted to simultaneously detect the presence of novel coronavirus 2019-nCoVand an internal reference gene Rnase P, and can also detect the presence of the RNA of the novel coronavirus 2019-nCoV in specimens such as bronchoalveolar lavage fluid, nose swabs and throat swabs.Through the primer, probe, kit and method, detection time is short and can be shortened to 1.5 h, so that the primer, probe, kit and method are suitable for rapid clinical and bedside detection and diagnosis; and the primer, probe, kit and method are high in detection of virus specificity, and high in repeatability, sensibility and accuracy rate.

The invention relates to the technical field of biological detection, and particularly discloses a primer and probe composition for detecting helicobacter pylori 23S rRNA gene mutation, application of the primer and probe composition and a kit. The primer and probe composition comprises a mutant primer and probe composition for detecting an A2142C site, an A2142G site and an A2143G site of a helicobacter pylori 23S rRNA gene, a primer and probe composition for detecting a helicobacter pylori 16S rRNA gene, a primer and probe composition for detecting a human RNAse P gene, and a blocking primer. The primer and probe composition is applied to preparation of a reagent or a kit for detecting helicobacter pylori 23S rRNA gene mutation. The kit comprises the primer and the probe composition. According to the application, the detection efficiency of helicobacter pylori 23S rRNA gene mutation is improved.

The invention relates to the field of molecular biology, in particular to a circular RNA targeted knockdown method of and application thereof. According to the circular RNA knockdown method, dCas13 protein is used for fusion expression of HRSP12, dCas13 and matched gRNA are used for specific targeting of circular RNA, RNase P/MRP compounds are recruited through dCas13-HRSP12 fusion protein, specific targeting degradation of circular RNA is achieved, and the method has the beneficial effects that interference to in-vivo steady-state environment is little, experiment results are more real and reliable, and specific targeting knock-down efficiency is high.

The invention provides a novel coronavirus fluorescent PCR detection kit and a use method thereof. The novel coronavirus fluorescent PCR detection kit comprises primers, probes, Taq DNA polymerase, UNG enzyme, dNTPs, Tris-HCl, MgSO4, glycerol and gelatin, wherein the primers and the probes are used for detecting a novel coronavirus ORFlab gene and an N gene and an internal standard virus RNase P gene; the probes of the ORFlab gene, the N gene and the RNase P gene are all probes modified by locked nucleic acid. The detection limit of the kit is 200 copies/mL, the locked nucleic acid probe, UNG enzyme, glycerol and gelatin are adopted, false negative results can be effectively avoided, and the kit has the advantages of being good in specificity, high in sensitivity and good in reproducibility.

The invention provides primers, probes and a detection method for nucleic acid detection of 2019-nCov or SARS-Cov-2 virus, including upstream and downstream primers and probes used for amplifying Orf1ab, E and N regions covering 2019-nCov virus nucleic acid, and upstream and downstream primers and probes used for detecting internal standard gene RNase P. The four probes are labeled by different fluorophores, and all detection can be completed in one tube through condition optimization, so that the detection cost and the detection time are greatly saved. The detection reagent and the detectionmethod have the advantages of good detection specificity, high sensitivity and simple operation, and can realize rapid and accurate detection of 2019-nCov or SARS-Cov-2 viruses.