33 results about "INHA" patented technology

The invention relates to a method for detecting multi-drug resistance of Mycobacterium tuberculosis, which aims at detecting the resistance of the Mycobacterium tuberculosis to isoniazid and rifampicin at the same time and has the characteristics of high specificity and sensitivity, quickness in detection, and easiness and convenience in operation. The technical scheme is as follows: the method comprises the following steps: A, establishing PCR (polymerase chain reaction) templates of an MTB clinical strain and a standard strain H37Rv; B, designing three pairs of primers of katG, inhA and rpoB gene fragments which are closely related to the isoniazid resistance and the rifampin resistance, performing mPCR amplification on katG, inhA and rpoB genes of the clinical strain, and performing PCR amplification on katG, inhA and rpoB genes of the clinical strain and the standard strain H37Rv; and C, detecting the mutation conditions of related INH-resistant and RFP-resistant 3 genes of the clinical strain at the same time by a single-strand conformation polymorphism detection method.

Compositions and process are provided for the rapid and specific detection of drug resistant forms of Mycobacterium tuberculosis based on real time PCR and high resolution melt analysis. The compositions and processes are useful for the detection of mutations within the Rifampicin Resistance Determinant Region (RRDR) of rpoB for the detection of rifampicin (RIF) and within specific regions of katG and the inhA promoter for the detection of isoniazid (INH) resistance. The invention also is capable of rapidly discriminating Mycobacterium tuberculosis complex (MTBC) strains from Nontuberculous Mycobacteria (NTM) strains.

A drug-resistance gene film chip for detecting mycobacterium tuberculosis is prepared by designing 54 probe spotting on a nylon film by aiming at the mutant sites of mycobacterium tuberculosis rpoB, kagG, embB, inhA, ahpC, gyrA, rrs, rpsL and pncA gene; 12 pairs of specific primers with biotin-labeled at 5' end are utilized; a sample DNA is subjected to triple PCR and amplified to form a large amount of gene fragment products with biotin; the amplified products and the probes on the film chip carry out specific hybridization; and then film washing, enzyme-linking and color reaction are carried out, thus preparing the chip. The gene film chip and the detection method thereof can detect the common gene mutations of the mycobacterium tuberculosis on the drug resistance of drugs such as isoniazid, rifampicin, streptomycin, ethambutol, pyrazinamide, quinolone and the like at one step, and are applicable to extracorporeal detection sputum sample, clinical isolation strains and mycobacterium tuberculosis multi-drug resistant gene in the organization sample.

The invention belongs to the field of biotechnological detection, and particularly relates to a multidrug-resistant mycobacterium tuberculosis detection method and a kit. The kit comprises PCR (polymerase chain reaction) forward and reverse amplification primers (a biotin label is arranged at the 5' end of the forward amplification primer) of the nucleic acid segments of the major drug-resistant gene of mycobacterium tuberculosis rifampicin and isoniazide and an oligonucleotides probe sequence (an amino mark is arranged at the 5' end); and the mutation conditions of the loca 511, 516, 526, 531 and 533 of the rpoB gene, the locus 315 of the katG gene and the locus 15 of the inhA gene can be detected at the same time in a reaction unit. The kit provided by the invention can quickly detect the mutation condition of the major drug-resistant gene of mycobacterium tuberculosis rifampicin and isoniazide, has low requirements on instrument and expenses, is easy to operate and can be used for detecting the drug-resistant gene of the mycobacterium tuberculosis.

The invention relates to the technical field of biological gene detection, and aims to provide an ovarian premature senility related gene whole exon amplification and detection method. The method comprises the following steps: extracting genome DNA from a detection material; using totally 120 primers shown in SEQ ID NO:1-120, carrying out an amplification test on all exons of 9 genes, namely FMR1,FOXL2, FSHR, POF1B, INHA, NOBOX, GDF9, BMP15 and FIGLA, wherein totally 61 PCR reactions in the amplification test and the negative control are carried out synchronously, and the Tm value range of the 120 primers is 57-62 DEG C; detecting an amplification product by adopting agarose electrophoresis and a gel imaging system; and analyzing the sequencing result of the amplification product by usingan automatic sequencer. All exons of 9 genes related to premature ovarian failure are synchronously amplified through optimized PCR amplification specificity, and a detection raw material is providedfor downstream detection. The method is simple and convenient to operate, short in amplification time, high in specificity and good in repeatability. The detected positive rate is greatly improved, and the application prospect is wide.

The invention discloses novel shRNA for increasing animal fertility and an application thereof, and the shRNA is characterized in that the shRNA comprises gene sequences of Sequence NO. 15 and Sequence NO. 16. According to the invention, several pairs of shRNA single-chain oligonucleotides are designed and synthesized based on the nucleotide sequence of sheep interferon INHA gene alpha subunit; an INHA shRNA interference vector pGFP-V-RS-shRNA is constructed; the interference effect is screened by a RT-PCR method so as to obtain a pGFP-V-RS-shRNA vector with good inhibition effect; the vector comprises gene sequences of Sequence NO. 15 and Sequence NO. 16; and a shRNA lentivirus interference vector pLenti6-shRNA3 is further constructed which comprises the gene sequences of Sequence NO. 15 and Sequence NO. 16. The invention also discloses an application of the novel vector.

A method to detect whether a female subject is predisposed to POF, the method comprising analysing at least one or more polymorphism in the INHA gene chosen from the group consisting: −124A>G; −16C>T; TG repeat (as herein after described); and one or more polymorphism in the INHA gene which is in linkage disequilibrium with one or more of −124A>G, −16C>T or TG repeat (as herein after described).

The invention provides a reagent kit and method for detecting an M.tuberculosis isoniazide drug tolerance mutation gene, particularly a multiplex PCR amplification system of three target genes katG, inhA and aphC with M.tuberculosis isoniazide drug tolerance mutation is established, and through a high-resolution melting curve method, wild types and different mutation types can be distinguished. Inaddition, through binding a molecular beacon probe, a multiple asymmetric PCR reaction can be realized, so that the mutation site of a tuberculosis resisting bacterial strain can be detected quicklyand accurately, and multiple sites can also be detected in a disposable manner.

The invention discloses primers and a kit for quickly detecting Mycobacterium tuberculosis isoniazide drug resistance. The primers comprises at least one set of the following primers: first set: katG Forward: 5'-TCGTATGGCACCGGAACC-3' and katG Reverse: 5'-CAGCTCCCACTCGTAGCC-3'; second set: katG-1 Forward: 5'-GGGCTGGAAGAGCTCGTAT-3' and katG-1 Reverse: 5'-CCGTACAGGATCTCGAGGAA-3'; and third set: inhA Forward: 5'-CGTTACGCTCGTGGACATAC-3' and inhA Reverse: 5'-TCCGGTAACCAGGACTGAAC-3'.

The invention discloses a gene editing system for high-quality porcine nuclear transplantation donor cells with high fecundity and resistance to porcine reproductive and respiratory syndrome and series diarrhea diseases and application of the gene editing system. The invention discloses a CRISPR/Cas9 system for pig INHA-CD163-pAPN gene editing. The CRISPR/Cas9 system comprises a Cas9 expression vector, a gRNA expression vector aiming at a pig INHA gene, a gRNA expression vector aiming at a pig CD163 gene and a gRNA expression vector aiming at a pig pAPN gene, the Cas9 expression vector is a plasmid complete sequence as shown in SEQ ID NO. 2. According to the invention, corresponding gRNA expression vectors are respectively designed aiming at different targets of INHA, CD163 and pAPN genes, and gRNA with relatively high editing efficiency and the expression vector thereof are obtained through screening. The modified Cas9 efficient expression vector is used for gene editing, and the editing efficiency is remarkably improved compared with that of an original vector.

The invention provides a mycobacterium tuberculosis rifampicin and isoniazide drug-resistant mutation detection kit and method, and particularly discloses a method and kit for detecting mycobacterium tuberculosis rifampicin and isoniazide drug-resistant mutation based on a fluorescent PCR melting curve method. The method and kit can be used for rapidly and qualitatively detecting drug-resistant mutations of drug-resistant determining regions of rifampicin drug-resistant genes rpoB and isoniazide drug-resistant genes katG, inhA and ahpC in a positive sputum culture sample of a mycobacterium tuberculosis complex of a tuberculosis patient in vitro.

Methods for the rapid detection of the presence or absence of Mycobacterium tuberculosis (MTB) resistant to rifampicin (MTB-RIF) and/or MTB resistant to isoniazid (MTB-INH) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the genes for rpoB, inhA, and katG, along with kits are provided that are designed for the detection of MTB-RIF and/or MTB-INH.

The invention provides a method and a chip for detecting multidrug-resistant mycobacterium tuberculosis. The method and the chip disclosed by the invention relate to detection of katG315 mutant I and/or II or rpoB526 mutant I and/or II, rpoB531 mutant I and/or II, and inhA-15 mutant. The invention also provides a primer for detecting the mutants.

The invention provides a method for preparing isoniazid nicotinamide adenine dinucleotide by synthetic biology, wherein the method includes the steps: 1) PCR amplification of katG and inhA DNA sequence fragments; 2) construction of a pTrcHis2c-katG plasmid; 3) construction of a pET28a-inhA plasmid; 4) purification of an InhA protein; and 5) purification of an INH-NAD compound. The method has the beneficial effects of directly synthesizing the INH-NAD in escherichia coli cells and realizing purification and separation of the INH-NAD through ultrafiltration tube concentration and purification, short-time heating, centrifugation, filtration and other steps.

The invention provides a method for detecting fecundity of alpine Merino sheep based on INHA gene, which relates to the technical field of genetic engineering, and detects the base of the first exon of INHA gene of alpine Merino sheep at 206bp . When the base is T, the genotype is TT; when the base is A, the genotype is TA or AA; the fecundity of the genotype AA is greater than that of the genotype TA, and the fecundity of the genotype TA is greater than that of the genotype TT. The invention can judge the fecundity of the alpine merino sheep by detecting the base at 206bp of the first exon ofthe INHA gene of the alpine merino sheep.

A method for database service to prevent dual master node using HAis provided, The method includes such steps as setting up local listening port in database service of host and standby in HA cluster system, monitoring local listening port before database instance is started, receiving connection of other database instance to local listening port, setting up local listening port in HA cluster system, setting up local listening port in HA cluster system, setting up local listening port in HA cluster system, setting up local listening port in HA cluster system, setting up local listening port inHA cluster system, setting up local listening port in HA cluster system, setting local listening port in HA cluster system, setting local listening port in HA cluster system, setting local listening port in HA cluster system. S2, in the dual-computer HA cluster system, the connection IP address of the opposite end server is respectively set in the database service of the host computer and the standby computer; S3, in the dual-machine HA cluster system, when the HA cluster resource manager starts the resources of the database service, the database service first listens to the monitoring port set locally for preventing the dual-master, and then connecting the monitoring port of the database service of the peer through the set peer server IP; S4, executing the startup process of the local database instance and other startup processes in the HA cluster system until the dual-machine HA cluster system is successfully started.

The invention relates to a drug-resistant site of mycobacterium tuberculosis and a detection method thereof. The invention discloses a mycobacterium tuberculosis drug-resistant gene locus, a primer group and a detection method based on a MassARRAY nucleic acid mass spectrum platform. The detection method comprises the following steps: designing amplification primers covering drug-resistant loci such as rpoB, inhA, katG, eis, rrs, gyrB and gyrA and extension primers covering drug-resistant loci such as rpoB, inhA, katG, eis, rrs, gyrB and gyrA; preparing a nucleic acid template, preparing a corresponding extension mixed reaction solution, and carrying out PCR amplification reaction to obtain a target product; and carrying out SAP enzyme digestion reaction and extension reaction: carrying out desalination treatment on the obtained extension amplification product, carrying out sample application, and analyzing by using analysis software of a mass spectrometer. The method is high in flux, good in flexibility, easy to implement and high in cost performance.

The invention discloses a nucleotide sequence of a recombinant-expression human inhibin A, and application of the nucleotide sequence. The application of the nucleotide sequence comprises the human inhibin A which is expressed by the nucleotide sequence, a preparation method for the recombinant-expression carrier of the human inhibin A, and a preparation method for the human inhibin A. The above method adopts artificial whole genes to synthesize the nucleotide sequence of the human inhibin A, a signal peptide nucleotide sequence, a leading peptide nucleotide sequence, a restriction enzyme cutting site, a G+C content, a purification tag and the like on the sequence are optimally designed to enable the nucleotide sequence to be more suitable for mammal cell expression; an INHA (inhibin A) gene sequence is synthesized by whole genes, and therefore, stability is good; a Kozak sequence is designed, expression can be accelerated, and the yield of the inhibin A is improved; and a His tag is designed so as to be favorable for purifying the expressed inhibin A.

The present invention discloses a kit for prenatal screening during pregnancy, the kit comprises magnetic beads coated with a production screening polyclonal antibody 1, an analysis buffer solution and an enzyme working solution, the production screening polyclonal antibody 1 is a mixture of AFP, Total beta hCG, PAPP-A, INHA and uE3 monoclonal antibodies 1, the enzyme working solution comprises anenzyme labeled production screening polyclonal antibody 2, and the production screening polyclonal antibody 2 is a mixture of AFP, Total BetahCG, PAPP-A, INHA and uE3 monoclonal antibodies 2. According to the invention, the magnetic beads are coated with the sieve-producing polyclonal antibody 1 and the sieve-producing polyclonal antibody 2 is marked on the enzyme to form an enzyme working solution, and the polyclonal antibodies in the sieve-producing polyclonal antibody 1 and the sieve-producing polyclonal antibody 2 are mixtures of AFP, Total beta hCG, PAPPA, INHA and uE3, so that the sensitivity of the kit can be obviously improved, and the usage is more convenient.