111 results about "Pcr test" patented technology

The present invention relates to fluorescent probe, and is especially fluorescent probe for real-time PCR detection and capable of resisting high temperature polyase exonuclease activity. The fluorescent probe has two-stage structure containing complementary sequences and including replacing probe, molecular beacon probe, scorpion primer, etc. The fluorescent probe includes a fluorescent probe such modified that it can resist the 5'-->3' exonuclease activity of high temperature polyase without generating non-specific signal, a fluorescent probe such modified that it can resist the 3'-->5' exonuclease activity of high temperature polyase without generating non-specific signal in real-time PCR detection, and a fluorescent probe such modified that it can resist both 5'-->3' exonuclease activity and 3'-->5' exonuclease activity of high temperature polyase without generating non-specific signal.

The present invention relates to a real time fluorescence metered PCR test way of HBV and a reagent kit. The present way adopts alkali cracking to extract HBV DNA, gets and adds a portion of the trace lysate directly into PCR reaction liquid for augmentation after cracking procedure; the reaction liquid includes a pair of specificity primers and a probe marked with fluorescent dye, PCR method is applied to augment target genes, the intensity of the reaction system is real-time tested, and Ct value is calculated according to analysis software as well as the virus content value of specimen. Under the conditions that sensitivity and accuracy are guaranteed, the reagent kit of the present invention can be operated easily and fast, thus reducing the labor intensity of clinical test and the cost of clinical PCR diagnosis at the same time, and continuing the rapidness and easiness function of fluorescence metered PCR test way. The way which is suitable for the clinical HVB metered test can be used as an assistant diagnosis way of HBV infection and a monitor mean of clinical treatment effect in a clinical lab, has comparatively low requirements of test operator, and possesses practical clinical application value.

The invention provides an extraction-free method and a kit for qualitatively detecting SARS-CoV-2 by fluorescence polymerase chain reaction (fluorescence PCR), and belongs to the field of biotechnology and medicine. The method integrates sample nucleic acid release and amplification. The kit comprises a lysis solution and a rapid enzyme, can directly lyse and amplify respiratory secretions of a patient, make a whole detection process fast and simple, reduce chance of mutual contamination, obtain accurate and reliable PCR test results, can diagnose presence of SARS-CoV-2 in patient secretions as soon as possible, and has important application value in the fields of disease monitoring and clinical diagnosis.

Thisa invention relates to a semi-set RT-PCR test reagent box for testing and verifying wheel-like virus including a MLV inverse transcriptase, Rnasin, DNA poly-enzyme, primers, dNTP, MgCl2 and 10 x buffer solution, in which, the upstream primer P1 is 5'-GTA TGG TAT TGA ATA TAC CAC-3', the downstream primer P2 is 5'-GAT CCT GTT GGC CAT CC-3. The RT-PCR test method includes: picking up a due test sample template, adding MLV inverse transcriptases, Rnasin, DNA poly-enzymes, primers, dNTP, MgCl2, 10 x buffer solution, the due test sample template and DECP water to be mixed uniformly to be enlarged on a PRC instrument, then the product is electrophoresis-enlarged in a device to get a result, which is recorded, analyzed and judged.

The invention relates to a green smut germ real time fluorescence fix quantity PCR agent box and the application. The box uses Taqman and positive primer, negative primer and masculine standard product to build up real time fluorescence PCR reacting system that could rapidly, and quantified testing green smut germ. It is suited for farm environment. It is also could be used to testing the germ testing for paddy seed and the multiplication speed in different paddy kinds.

The invention relates to a PCR test method of a vibrio parahaemolyticus with internal amplification control in the food safety technology filed, which comprises steps as follows: firstly the construction and synthesis of the internal amplification control sequence artificially and design of the specific primers; secondly the PCR reaction system is laid in the best reaction condition; thirdly PRC is amplified; fourthly DNA of the different-serotype vibrio parahaemolyticus and different-species non vibrio parahaemolyticus is extracted for PRC test to identify the specific primers and confirm whether the internal amplification control can be normally amplified in the non vibrio parahaemolyticus sample test; finally the evaluation of the built PCR reaction through the artificial and natural pollution samples tests. The invention shows the occurrence of the false negative in the testing process and improves the accuracy rate. The invention provides efficient and reliable technical means to satisfy the urgent need of the foodborne patheogen investigation and test in the quarantine law enforcement.

The invention relates to a preparation method of a multiple amplification internal standard used for the PCR detection of four bacteria of the technical field of food safety, the steps are that: a specific detection primer is designed according to a staphylococcus aureus vicK gene, a salmonella invA gene, a listeria monocytogene hlyA gene and vibrio parahaemolyticus toxR gene; the invention adopts an overlap PCR technology for assembling the sequence of the specific detection primer of the staphylococcus aureus vicK gene, the salmonella invA gene, the listeria monocytogene hlyA gene and the vibrio parahaemolyticus toxR gene into a nucleotide segment so as to construct the multiple amplification internal standard; the artificial sequence is cloned to a plasmid vector pMD-18T to constitute a plasmid pMD-mIAC which comprises the sequence of the multiple amplification internal standard; the multiple amplification internal standard is respectively verified in the detections of staphylococcus aureus, salmonella, listeria monocytogenes and vibrio parahaemolyticus. The invention can greatly reduce the detection cost and ensure the accuracy and the high efficiency of the detection at the same time.

The invention discloses a wild ginseng and cultivated ginseng multiple PCR test kit, which is characterized by containing: buffer solution, 12.5mM of deoxy-ribonucleoside triphosphate (dNTP), 0.1mM of one of a primer 1 and a primer 2, Taq DNA polymerase, a sample DNA to be tested and a double-distilled water identification reaction system; or the buffer solution, 12.5 mM of dNTP, 0.1 mM of one of the primer 1 and the primer 2, Taq DNA polymerase, wild ginseng and cultivated ginseng DNA 1:1 mixture and a double-distilled water positive reference reaction system; or the buffer solution, 12.5 mM of dNTP, 0.1 mM of one of the primer 1 and the primer 2, Taq DNA polymerase, araliaceae congeneric DNA 1:1 mixture, and a double-distilled water negative reference reaction system. The detection and identification method comprises the steps of designing two pairs of specific oligonucleotide primers of wild ginseng and cultivated ginseng mitochondrion DNAs, designing two pairs of specific oligonucleotide primers of synthetic wild ginseng and cultivated ginseng mitochondrion DNAs, determining a reaction process, determining result and the like. The method can accurately determine the specificity of both the wild ginseng and the cultivated ginseng, and the detection result is reliable.

This invention provides a primer combination of melon anthracins used in a specific test sample and a PCR test reagent box containing said primer combination and a method for testing melon anthracins.

This invention relates to a fluorescence quantitative PCR reagent box and a method testing vibro parahaemolyticus. The reagent box is composed of PCR reacting agent tube, the tube of sample treatment, the tube of positive comparison, the tube of negative comparison, the tube of standard product, the tube of Taq DNA polyose and the tube of dehydronium. The testing procedures are: first collect and treat the pending tested sample, then implement molecular quantitative PCR test, according the CT values of positive and negative sample comparisons and the CT of tested samples to judge whether the sample is positive or negative, then record the fluorescence value and CT value of each sample, and get the standard curve from the computer to calculate the copy's number of vibro parahaemolyticus of each sample. This invention has sealed-loop operation, avoids the fake positive, and the reaction and hybridization is synchronous, and the testing time is short.

The present invention relates to microbe detecting composition and method, and is especially PCR kit and test method of testing banana apical tuft virus. The PCR test kit includes primer, dNTPs, buffering solution and DNA polymerase, and its upstream primer is 5-CTC GTC ATG TGC AAG GTT ATG TCG-3' and downstream primer is 5'-GAA GTT CTC CAG CTA TTC ATC GCC-3'. The PCR test kit is easy to produce and the test method of testing banana apical tuft virus has high sensitivity and high repeatability and is simple and practical. In detecting coarsely purified virus liquid, the method can detect banana apical tuft virus equivalent to the amount of 1 ng of banana tissue in high test precision and relatively low test cost.

The invention relates to the technical field of biology, in particular to a reagent which can simultaneously distinguish and detect four animal insect-borne diseases as well as a preparation method and application of the reagent. The animal insect-borne disease multi-RT-PCR distinguishing and detecting reagent comprises four pairs of specific primers, and the respective amplification target fragment lengths of Bluetongue virus (BTV), epizootic haemorrhagic disease virus of deer (EHDV), Vesicular stomatitis virus (VSV) and Akabane virus (AKV) are 351bp, 536bp, 300bp and 250bp. The VP7 of the BTV, the EHDV, the VSV and the AKV and a conservative fragment of an N gene are respectively selected as targets, and the primer Express software and the primer prere 5.0 software are applied to deign and combine a primer. An optimal matching and screening test and a multi-RT-PCR test are carried out on a plurality of pairs of designed primers, and four pairs of primers which can carry out the distinguishing and detection on the four animal insect-borne diseases and have high amplification efficiency and good specificity can be obtained by a plurality of reaction condition optimization and comparison tests and verification tests. A kit formed by the reagent can obtain a qualitative distinguishing and detecting result in six hours after a sample is received. The multi-RT-PCR distinguishing and detecting reagent is a sensitive and reliable method for detecting the BTV, the EHDV, the VSV and the AKV in the clinical sample.

This invention provides a PCR test reagent box for diagnosing and testing An- masked sporidiosis, which is prepared by designing specific primers according to the ITS-1 gene sequence of the masked sporidiosis, setting up PCR test method and optimizing the PCR reaction condition. This invention also provides a method for using the box.

The invention relates to a method for transforming Sudan grass through an agrobacterium mediating gene, which belongs to the biotechnology field. When the agrobacterium mediating method is used to transforming exogenous gene of Sudan grass, a faint yellow granular callus which is obtained from the in-vitro culture and induction of a Sudan grass young ear is taken as acceptor material; after the infection and co-culturing of the agrobacterium , the acceptor material undergoes 10 to 20mg/L kanamycin resistance screening to obtain a resistance callus in a subculture medium of carbenicillin with attached antibiotics of 500mg/L; the resistance callus is translated into a differentiation culture medium of the carbenicillin with the attached antibiotics of 500mg/L to obtain a regenerated plantlet; the plantlet is cultured to have sound seedling and root and survives after transplantation; and a transgenic plant is obtained after the plant passes a PCR test. The Sudan grass genetic transformation method established by the invention has a transformation rate of more than 3 percent, thereby laying the foundation for carrying out the transgenic breeding of the multi-purpose Sudan grass through the agrobacterium mediating method.

The upstream pimer for testing sulfate reducing bacterium is 5-CGCTGAAATGACCATGATGG-3, and its downstream primer is 5-CGCGCATTTCACGAAGC-3. The fluorescent probe is composed of reporter fluorescent group, quenching fluorescent group and probe gene sequence, which being of 5-CTGAAGCCTTACGAAGATCG-3. Said 5 end of probe gene sequence labels that reporter fluorescent group is FAM; and said 3 end of probe gene sequence labels that quenching fluorescent group is TAMRA. This invented primers and fluorescent probe have advantages: high specificity; short-term testing of sulfate bacterium for fluorescent quantitative PCR test, and needing only 6 Hrs. after DNA extraction and fluorescent quantitative PCR test.

This invention belongs to the moleculal biology field. It involves a kind of fluorescent quantitive PCR test kit of detection gene mutations. The test kit includes fluorescent quantitive PCR premixed liquid, fluorescent quantitive reaction liquid A, luorescent quantitive reaction liquid B (the feature is that it contains primer and anti-primer and fluorescence probes), positive comparison sample A, positive comparison sample B, positive comparison sample C. This invention features that: short detection time, easy operation, low quality requirement of sample DNA, the result reading is clear, direct with high sensitivity.

The invention relates to a test method in the bioengineering field, in particular to a fluorescence quantitative PCR test method used for quantitatively test the titer of Hepatitis E virus (HEV) in a clinical specimen. The steps of invention include the design of virus specific quantitative PCR primers and probes, the construction of standard molecular, the extraction of a virus sample RNA, the cDNA synthesis of the sample RNA and the establishment and optimization of the fluorescence quantitative PCR test method. The testing sensitivity of the invention can reach to 10 degrees magnitude virus copy. The invention is equipped with good stability and specificity, which can be used for mass samples test and is capable of precisely measuring the virus load of the sample.