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74 results about "Murine leukemia virus" patented technology

The murine leukemia viruses (MLVs or MuLVs) are retroviruses named for their ability to cause cancer in murine (mouse) hosts. Some MLVs may infect other vertebrates. MLVs include both exogenous and endogenous viruses. Replicating MLVs have a positive sense, single-stranded RNA (ssRNA) genome that replicates through a DNA intermediate via the process of reverse transcription.

Heat-resistant reverse transcriptase mutant

Provided are: a reverse transcriptase mutant including an amino acid mutation at a position corresponding to position 55 of the amino acid sequence of wild-type reverse transcriptase derived from the Moloney murine leukemia virus, wherein the reverse transcriptase mutant is characterized in that the amino acid mutation is a substitution from threonine to another amino acid, and the other amino acid is selected from the group consisting of amino acids having a nonpolar aliphatic side chain and amino acids having a polar acidic functional group side chain; a nucleic acid that encodes the mutant; a method for producing the mutant and the nucleic acid that encodes the mutant; a method for synthesizing cDNA in which the mutant is used; and a composition and kit including the mutant.
Owner:TAKARA HOLDINGS

Avian influenza H7N9 virus RT-PCR (reverse transcription-polymerase chain reaction) detecting kit and detecting method

InactiveCN103276109AStrong specificityMeet the needs of prevention and control in a timely mannerMicrobiological testing/measurementMicroorganism based processesFluorescenceReverse transcriptase
The invention relates to an avian influenza H7N9 virus RT-PCR (reverse transcription-polymerase chain reaction) detecting kit and a detecting method, and aims at providing the detecting kit which has the characteristics of convenience in use and accuracy in detection, and the detecting method which has the characteristics of high accuracy, simplicity and convenience in detection. The technical scheme is as follows: the avian influenza H7N9 virus fluorescence-quantitative RT-PCR detecting kit comprises deoxynucleotide triphosphate, MgCl2, an RT-PCR buffer solution, an avian influenza H7N9 virogene standard product, an RNA (Ribonucleic Acid) enzyme inhibitor, an MMLV (Moloney Murine Leukemia Virus) reverse transcriptase and a DNA (Deoxyribonucleic Acid) polymerase, and is characterized in that the detecting kit also comprises an upstream primer, a downstream primer and a specific probe. The avian influenza H7N9 virus fluorescence-quantitative RT-PCR detecting method comprises the following steps of: (1) extracting RNA of a sample to be detected; (2) carrying out RT-PCR reaction; and (3) carrying out fluorescence detection on the RT-PCR reaction product.
Owner:ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION

Method for preparing visual gene detection reagent based on G4DNAzyme coloration

The invention relates to a method for preparing a visual classical swine fever virus (CSFV) gene detection reagent based on G4DNAzyme coloration. The method comprises the following steps: selecting cells infected with viruses, after freeze thawing and lysing, adopting a gene extraction kit for extracting a target gene and obtaining cDNA after carrying out reverse transcription and inactivation onMoloney murine leukemia virus (MMLV) reverse transcriptase; adding the cDNA to an asymmetric polymerase chain reaction (PCR) system and obtaining an asymmetric PCR product through degeneration, annealing and extension amplification for 50-100 cycles; and adding upstream and downstream probes and the asymmetric PCR product to G4DNAzyme coloration reaction buffer, and after degeneration and annealing, adding Hemin, ATBS and H2O2 to carry out coloration reaction and observing whether macroscopic green color appears, thus judging whether CSFV infection exists. The method has the beneficial effects of high detection speed, accuracy, stability, good repeatability, simple and easy-to-operate detection steps, capability of directly observing the coloration reaction with naked eyes, intuitionisticresults and low cost.
Owner:巨星农牧有限公司 +1

LAMP (Loop-Mediated Isothermal Amplification) detection method of PRV (Porcine Rotavirus) inverse transcription and application

The invention discloses an LAMP (Loop-Mediated Isothermal Amplification) detection method of PRV (Porcine Rotavirus) inverse transcription and an application. The detection method comprises the following steps: (1) building a RT-LAMP reaction system, and setting a negative control at the same time, wherein the RT-LAMP reaction system is 25 [mu] L and comprises 2 [mu] L of template, 0.5 [mu]L of 0.8 [mu]M FIP primer, 0.5 [mu]L of 0.8 [mu]M BIP primer, 0.25 [mu]L of 0.2 [mu]M of F3 primer, 0.25 [mu]L of 0.2 [mu]M of B3primer,2[mu]L of dNTP (Diethyl-Nitrophenyl Thiophosphate), 5 [mu]M of MgSO4 (Magnesium Sulfate), 8UBst of DNA polymerase and 1*ThermoPol Buffer, and 1[mu]L of MLV (Murine Leukemia Virus) reverse transcriptase, and supplementing the volume to 25 [mu]L by sterile water; and the negative control template is sterile water; (2) after reacting for 20-60min at 61-65 DEG C in a constant temperature water bath boiler, terminating for 20 min at 80 DEG C, carrying out AGE (Agarose Gel Electrophoresis) identification on an amplification product or adding 2 [mu]L of SYBR Green dye, and observing a result in an UV lamp (Ultraviolet Lamp) or by naked eyes. According to the LAMP detection method provided by the invention, a convenient, fast and accurate molecular biological diagnosis method can be provided for the clinical diagnosis and epidemiological investigation of the PRV.
Owner:NORTHEAST AGRICULTURAL UNIVERSITY

Method for detecting nucleic acid of porcine reproductive and respiratory syndrome virus in one step

The invention relates to a method for detecting nucleic acid of a porcine reproductive and respiratory syndrome (PRRS) virus by one step, which comprises the following steps of: collecting, processing and detecting samples, wherein in the collecting step, an animal blood sample is dripped into a full type approval (FTA) card sample area; and the detecting steps comprises the: (1) designing specific primers and a probe for general type PRRS virus, and marking carboxyfluorescein (FAM) and tetramethyl rhodamine (TAMARA) fluorescent groups on the probe; and (2) preparing and optimizing a detection system and a reaction condition, wherein a reaction system comprises 25 or 50 microliters of trihydroxymethyl aminomethane-hydrogen chloride (HCl) (the pH value is between 7.8 and 9.0), 0.1 to 0.5 micro mol of upstream primer and 0.1 to 0.5 micro mol of downstream primer, 100 to 400 micro mols of deoxynucleotide mixture, 0.1 to 0.5 micro mol of probe, 100 to 300 U of Moloney murine leukemia virus (M-MLV) reverse transcriptase, 1 to 5 U of thermostable deoxyribonucleic acid (DNA) polymerase, 4 to 8 mols of Mg<2+->, 300 to 500 nano mol of homogenized reference dyes ROX, and 1 to 15 microliters of sample which is re-suspended in trihydroxymethyl aminomethane-ethylene diamine tetraacetic acid (EDTA) buffer solution and is added before each time of reaction. In the method, the sample collection is easy and convenient, so that an FTA card containing s ribonucleic acid (RNA) sample can be posted to any one central laboratory to be detected according to a form of regular mails; and the pollution risks are reduced, and the detection sensitivity is improved.
Owner:湖南农安生物技术有限公司

Avian influenza virus H7 type RT-PCR (reverses transcription-polymerase chain reaction) detecting kit and detecting method

The invention relates to an avian influenza virus H7 type RT-PCR (reverses transcription-polymerase chain reaction) detecting kit and a detecting method, and aims at providing the detecting kit which has the characteristics of convenience in use and accuracy in detection, and the detecting method which has the characteristics of high accuracy, simplicity and convenience in detection. The technical scheme is as follows: the avian influenza virus H7 type fluorescence-quantitative RT-PCR detecting kit comprises deoxynucleotide triphosphate, MgCl2, an RT-PCR buffer solution, an avian influenza virus H7 gene standard product, an RNA (Ribonucleic Acid) enzyme inhibitor, an MMLV (Moloney Murine Leukemia Virus) reverse transcriptase and a DNA (Deoxyribonucleic Acid) polymerase, and is characterized in that the fluorescence-quantitative RT-PCR detecting kit also comprises an upstream primer, a downstream primer and a specific probe. The avian influenza virus H7 type fluorescence-quantitative RT-PCR detecting method comprises the following steps of: (1) extracting RNA of a sample; (2) carrying out RT-PCR reaction; and (3) carrying out fluorescence detection on the RT-PCR reaction product.
Owner:ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION

Detection kit and detection method of reverse transcription polymerase chain reaction (RT-PCR) of avian influenza virus N9

The invention relates to a detection kit and a detection method of reverse transcription polymerase chain reaction (RT-PCR) of an avian influenza virus N9, and aims at providing the detection kit and the detection method, wherein the provided detection kit has the characteristics of being convenient to use and accurate to detect; and the provided method has the characteristics of being high in detection accuracy, and simple and convenient to detecting process. The technical scheme is as follows: the detection kit of fluorescent quantitative RT-PCR of the avian influenza virus N9 comprises deoxy triphosphate nucleoside, MgCl2, RT-PCR buffer solution, an avian influenza virus N9 genetic standard, a ribonucleic acid (RNA) enzyme inhibitor, moloney murine leukemia virus (MMLV) reverse transcriptase and deoxyribonucleic acid (DNA) polymerase. The detection kit of fluorescent quantitative RT-PCR is characterized by also comprising an upstream primer, a downstream primer and a specific probe. The detection method of fluorescent quantitative RT-PCR of the avian influenza virus N9 is carried out according to the following steps of: (1) extracting the RNA; (2) carrying out RT-PCR; and (3) carrying out fluorescence detection on an RT-PCR reaction product.
Owner:ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION

Visual gene detection reagent for classical swine fever virus based on G4DNAzyme coloration

The invention relates to a method for preparing a visual classical swine fever virus (CSFV) gene detection reagent based on G4DNAzyme coloration. The method comprises the following steps: selecting cells infected with viruses, after freeze thawing and lysing, adopting a gene extraction kit for extracting a target gene and obtaining cDNA after carrying out reverse transcription and inactivation onMoloney murine leukemia virus (MMLV) reverse transcriptase; adding the cDNA to an asymmetric polymerase chain reaction (PCR) system and obtaining an asymmetric PCR product through degeneration, annealing and extension amplification for 50-100 cycles; and adding upstream and downstream probes and the asymmetric PCR product to G4DNAzyme coloration reaction buffer, and after degeneration and annealing, adding Hemin, ATBS and H2O2 to carry out coloration reaction and observing whether macroscopic green color appears, thus judging whether CSFV infection exists. The method has the beneficial effects of high detection speed, accuracy, stability, good repeatability, simple and easy-to-operate detection steps, capability of directly observing the coloration reaction with naked eyes, intuitionisticresults and low cost.
Owner:巨星农牧有限公司 +1
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