33 results about "Reverse dot blot" patented technology

The invention provides a kit capable of simultaneously detecting 6 deletion type alpha-thalassemia genes (-alpha<3.7>, -alpha<4.2>, --<SEA>, --<THAI>, --<FIL> and -alpha<27.6>) and 6 non-deletion type alpha-thalassemia genes (alpha<CS>alpha, alpha<QS>alpha, alpha<WS>alpha, alpha<13>alpha, alpha<43-44>alpha and alpha<49>alpha).The kit comprises a DNA chip, PCR reaction liquid I and PCR reaction liquid II, wherein the DNA chip comprises a substrate and probes fixed on the substrate.The kit has the advantages that the detection principle of PCR-reverse dot blot is used, the corresponding amplification primers and probes are designed according to the mutated or deleted sites of the genes, the probes are fixed on the DNA chip, and thalassemia is diagnosed through the hybridization of PCR products amplified by the specific primers and the probes fixed on the DNA chip and the interpretation of a signal developing box.

The invention relates to a kit for detecting a hepatitis c virus genotype, particularly relates to that the nucleic acid reverse dot blot hybridization technology is used to prepare a kit for hepatitis c virus genotype detection. The invention is used for rapidly and accurately distinguishing the hepatitis c virus genotype in a clinical blood sample.

The present invention disclosed the use of single nucleotide polymorphism (SNP) as the detection assay for human identification. Using the reversed dot-blot format and the flow through hybridization process, the process can be more efficient, less expensive and with similar or better power of exclusion in definitive identification. The present method can be applied to any other organisms.

The invention relates to a high throughput test method for a tomato bacterial disease, and more particularly relates to a high throughput test method which can utilize a locking probe to carry out reverse dot blot hybridization on tomato bacterial wilt original bacteria, tomato ulcer pathogenic bacteria and a tomato spot disease, and belongs to the crop preventing and curing disease and exit and entry plant quarantine range. The high throughput test method provided by the invention is based on a rolling circle amplification technology of the locking probe, a pair of universal primers are utilized for amplifying multiple pathogens, then, the high throughput test method and a reverse dot blot are combined together, a result is judged through a chromogenic reaction, the testing time of the traditional tomato bacterial disease is shortened, the specificity and the sensitivity of the detection are guaranteed, and the high throughput test is realized. According to the method, the testing cycle of the traditional pathogenic bacteria can be shortened, the cargo clearance can be increased, the intercept and capture relevance ratio of epidemic situations of imported bulk farm-product is improved, the pest invasion capacity is improved, the cargo clearance of cargoes imported and exported at ports in China is accelerated, and the enterprise cost is reduced and the high throughput test method has the important meanings.

The invention relates to examination mutant hepatitis virus method, especially the method of using DNA reverse dot blot hybridization technique to quickly and exactly distinguish clinic blood sample wild type and rummy fuding tolerance mutant hepatitis b virus. And it also relates to clinic measuring kit.

The invention relates to the technical field of biological detection, in particular to HPV (human papilloma virus) typing detection primers as well as a detection method and an application thereof. QPCR primers for typing detection of HPV are shown as SEQ ID NO:1-38; the invention also discloses an application of the qPCR primers for typing detection of the HPV in preparing an HPV detection kit. The specificity and the sensitivity of the primer pairs and the kit provided by the invention in HPV typing detection are greatly higher than that of a common reverse dot blot method, and the primer pairs and the kits are relatively low in requirements on environment and operation and are broad in application. Meanwhile, according to the invention, a DNA double-stranded chimeric fluorescent dye qPCR detection method is applied to the HPV typing detection; the detection specificity is improved based upon double analysis through an amplification curve and a melting curve, so that the specificity can reach the detection level of a probe method, and meanwhile, cost can be greatly reduced; the primers have an important significance for the promotion of HPV general survey and for the prevention and treatment of cervical carcinoma; and the primers are quite high in popularization and application values.

The invention relates to a method and a kit for detecting mutations in the pre-C region and core promoter region (BCP) of the hepatitis B virus (HBV) genome in clinical blood samples, in particular to rapid detection of the pre-C/BCP region of the HBV gene by reverse dot hybridization technology The method of mutation and the kit used.

The invention relates to a gene chip, a primer set and a kit for single-tube detection of non-deletion alpha-thalassemia and beta-thalassemia, and belongs to a molecular diagnosis technology. According to the present invention, based on the direct multiplex PCR and reverse dot blot combined detection principle, the corresponding amplification primers and the corresponding probes are designed according to the mutation or deletion sites of each genotype, the primer is labeled with biotin, the probe is labeled with amino, a gene chip is used as substrate, the probe is immobilized on the DNA chip,the PCR product amplified by the specific primer is hybridized with the probe, and the diagnosis of thalassemia is performed by interpreting a signal coloring box.

The invention discloses a probe for classificatory diagnosis of four plasmodia infecting human, a kit and a using method thereof, wherein species special probes are respectively adopted for plasmodium falciparum, plasmodium vivax, plasmodium ovale and plasmodium malariae; 10 T tails are added at the 5' end of the probe and are cross-linked and fixed on a nylon hybrid membrane in an ultraviolet manner. The kit consists of PCR and reverse dot blot hybridization reaction reagents and materials and can be used for classificatory diagnosis of plasmodium falciparum, plasmodium vivax, plasmodium ovale and plasmodium malariae or two or three or four of the plasmodium falciparum, plasmodium vivax, plasmodium ovale and plasmodium malaria, so that the invention provides a simple, convenient and quick plasmodium classificatory diagnosis technique.

The invention discloses a kit capable of detecting a natural water sample of prorocentrum donghaiense in a high-throughput manner. The kit comprises 12 constituents, including a natural sample nucleic acid crude-extract reagent, 5*NASBA (Nucleic Acid Sequence Based Amplification) Buffer, 5*Primer mix, 4*Enzyme mix, an RNA (Ribonucleic Acid) denaturing agent and a hybridization solution. The detection principle of the kit is as follows: rRNA (Ribosomal Robonucleic Acid) of an algae cell is taken as a target sequence which is prepared by using an isothermal nucleic acid amplification technology, and a plurality of detection samples are detected at the same time by virtue of the reverse dot blot of RNA. According to the kit, a detection process is in need of simple equipment only, the detection is rapid, a detection result can be judged directly through naked eyes, and the kit has high sensitivity, can be used for site detection of a daily water environment and has a certain practical value for early warning of the red tide of the prorocentrum donghaiense.

The invention relates to a method for specifically detecting and identifying retroviral nucleic acids/retroviruses in any item to be examined using RT-PCR and reverse dot blot hybridization (RDBH) as well as to diagnosis kits for carrying out said method. The invention also relates to retrovirus-specific, oligonucleotide/primer mixtures (MOP) which comprise forward primers and reverse primers and which are provided for generating amplification products of retrovirus-specific nucleic acids from the item to be examined. These inventive primary mixtures (MOP-ABD, MOP-C) are comprised of retrovirus-specific, degenerated oligonucleotides which correspond to the highly conserved regions located within the reverse transcriptase gene (RT-gene) of all known human retroviruses and which have a head or extension sequence consisting of a clamping and interface sequence. The invention also relates to retrovirus-specific probes for the RDBH, whereby defined quantities of synthetically produced, precisely defined nucleic acid sequences are concerned which stem from the reverse transcriptase gene of those retroviruses already characterized, against which the item to be examined should be tested, and which do not overlap at all with the nucleotide sequences of the forward primers and reverse primers used in the PCR or RT-PCR.

The invention provides an HPV typing detection method. The HPV typing detection method includes the following steps that an in-vitro sample to be detected and containing HPV is provided, and nucleic acid precipitant is adopted to extract nucleic acid according to different types of primers and hybridization probes marked by biotin and designed by L1 gene target sequence of HPV; the primers are used for PCR amplification of nucleic acid to obtain a PCR product with a nucleic acid fragment of an L1 region of HPV; the different types of hybridization probes are fixed to different positions of a hybridization chip and numbered to obtain a hybridization chip with the hybridization probes fixed; the PCR product and the hybridization chip with the hybridization probes fixed are subjected to reverse dot blot hybridization to obtain a nucleic acid chip; the nucleic acid chip is processed with streptavidin marked by alkaline phosphatase and then reacts with a chromogenic substrate BCIP/NBT for color development, and the type of HPV in the in-vitro sample to be detected is judged according to the chromogenic position of the nucleic acid chip.

The invention relates to a chip, an amplification reagent and a kit for directly and simultaneously detecting alpha-thalassemia and beta-thalassemia mutation sites, and belongs to a molecular diagnosis technology. According to the present invention, based on direct multiplex PCR, Gap-PCR and reverse dot blot combined detection principle, the corresponding amplification primers and the corresponding probes are designed according to the mutation or deletion sites of each genotype, the primer is labeled with biotin, the probe is labeled with amino, a gene chip is used as substrate, the probe is immobilized on the DNA chip, the PCR product amplified by the specific primer is hybridized with the probe, and the diagnosis of thalassemia is performed by interpreting a signal coloring box.

The invention discloses a method for detecting various respiratory viruses based on a gene membrane chip. The method comprises the following detection steps: pretreating a sample: taking a nasal/pharyngeal secretion swab into a preservation tube containing a sample preservation solution, fully washing, sucking all the solution into a new centrifuge tube, centrifuging, and discarding the supernatant; extracting nucleic acid: adding the extracting solution and protease K into a centrifugal tube, heating and centrifuging after vortex oscillation, and taking supernate for PCR; carrying out multiple PCR sample adding and amplification: taking multiple respiratory viruses PCR Mix, a nucleic acid extract solution and enzyme-free water, and carrying out multiple PCR sample adding and amplification; performing membrane chip hybridization: performing hybridization reaction on the multiplex PCR amplification product and a membrane chip through a full-automatic membrane chip nucleic acid molecule hybridization instrument; and issuing a detection result report: directly issuing the detection result report through an automatic analysis system of the full-automatic membrane chip nucleic acid molecule hybridization instrument. According to the invention, multiple PCR and reverse dot blot hybridization technologies are combined, so that multiple respiratory pathogens in a single sample can be simultaneously detected through a single reaction, and a detection result report can be automatically issued.

The invention provides a kit for detecting six common porcine viruses in a Fukuo raw material and a product thereof. The kit comprises a gene membrane chip, a hybridization reagent and an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) reaction system, the gene membrane chip comprises a nylon membrane substrate, and oligonucleotide probes, contrast probes and blank dot coatings which are distributed on the nylon membrane substrate in an array manner; the RT-PCR reaction reagent comprises a combination of seven primer pairs. Multiple PCR is combined with a reverse dot blot hybridization technology, seven specific primer pairs are utilized, seven specific gene segments can be amplified at the same time, and the specificity and accuracy of a detection result are further ensured through hybridization of PCR products and probes on a membrane chip; compared with common qualitative PCR and fluorescent quantitative PCR, the detection efficiency and accuracy are greatly improved.