33 results about "Reverse dot blot" patented technology

The present invention relates to a method and a reagent kit which are used for detecting the existence of the mycobacterium tuberculosis in the clinical biological sample and the mutations of the drug-resistant genes, in particular to a method and a reagent kit which are used for quickly detecting the existence of the mycobacterium tuberculosis in the biological samples such as the clinical sputamentum, the bronchoalveolar lavage fluid, the blood, the ascites, and the cerebrospinal fluid, etc. with the reverse dot-blot hybridization technology.

The invention relates to a kit for integrated detection of alpha and beta mutant type thalassemias, in particular to a kit for detecting alpha and beta thalassemias mutant types by using multiple asymmetric amplification and reverse dot blot hybridization techniques. The kit of the invention consists of a polymerase chain reaction (PCR) reagent, a low-density chip and a hybridizing reagent, and contains a set of specific nucleotide polymorphism probes and PCR primers for amplifying target gene in an amplification sample. The kit can be used for detecting six alpha thalassemia locus mutations and 27 beta thalassemia mutations, which are common in China, at the same time. The detection process of the kit is faster and the detection result of the kit is more accurate, so the kit is expected to be widely used in diagnosis of thalassemia in clinic and instruction on sound child rearing.

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 present invention discloses methods and devices for rapid genotyping. In one embodiment, the present invention is applied to human papillomavirus (HPV) genotyping, comprising the use of viral genotype-specific-oligonucleotide probes, reversed-dot-blotting genotype array format and flow through hybridization process, thereby providing a more efficient, faster and less expensive method for HPV genotyping. The genotyping method further comprises the use of generic probes to expand the detection of HPV genotypes.

The invention relates to a method and a reagent kit for detecting different gene subtypes of hepatitis b virus (HBV) in clinical blood samples and in particular relates to the method for rapidly detecting the HBV gene subtypes with a reverse dot blot hybridization technique and the reagent kid which is used in the method.

The invention relates to a helicobacter pylori specific molecular marker and a detection kit thereof. The kit is used for performing specific detection on helicobacter pylori in samples of gastric mucosa, gastric juice, excrement and throat swab according to PCR-reverse dot blot. A target gene at which the helicobacter pylori specific molecular marker aims is a urease ureH gene. With the adoptionof the kit, sensitivity of clinical examination of helicobacter pylori is improved greatly, and furthermore, helicobacter pylori infected persons, especially old people and children can be subjected to noninvasive screening, so that early detection and early treatment can be realized; besides, for detection of other helicobacter pylori genes such as cag and vec and other drug resistance genes, thekit can be taken as positive reference and used for monitoring all processes such as genome extraction, PCR and hybridization of other genes.

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 medical in-vitro diagnostic techniques, in particular to a reverse dot-blot hybridization gene chip. A plurality of microarrays in different areas are distributed on a nylon membrane, and a common specific probe for postoperative intracranial infection pathogenic bacteria is fixed in each microarray area. The reverse dot-blot hybridization gene chip can be used for identifying enterococcocci (enterococcus faecium and enterococcus faecalis), staphylococcus aureus, coagulase-negative staphylococci, escherichia coli, Klebsiella pneumonia, pseudomonas aeruginosa, acinetobacter baumannii and enterobacter cloacae. The reverse dot-blot hybridization gene chip further comprises a universal Gram-negative bacterium probe and a universal Gram-positive bacterium probe. The chip can be used for simultaneously detecting whether various pathogenic bacteria exist or not, has the advantages of rapidity and accuracy in diagnosis, high specificity and heavy information, and is of great help to clinical diagnosis and epidemic disease screening.

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 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 present invention discloses primers, a probe and a kit for reverse dot blot hybridization detection of mycobacterium tuberculosis and a usage method thereof. The kit is formed by a PCR primer group, a nylon hybridized membrane fixed with a mycobacterium tuberculosis-specific probe and other reagents for reverse dot blot hybridization. The primer group is constituted by an IS1' primer with the sequence of TACGGTGCCCGCAAAGTG and an IS2' primer with the sequence of AGGCGTCGGTGACAAAGG; and the probe comprises a Probe-B-3 'probe with the sequence of GCCTTTGTCACCGACGCCTA. The usage method of the kit is characterized by discriminating mycobacterium tuberculosis from other common pathogens at room temperature. The invention has the advantages of high sensitivity, strong specificity, simple operation and low temperature requirements, and provides technical support for discrimination and diagnosis of mycobacterium tuberculosis.

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.

The invention provides a method and a system for detecting brucella based on a micro-fluidic chip-PCR (Polymerase Chain Reaction) technology. The method for detecting Brucella comprises the following steps: performing target DNA extraction, PCR amplification and reverse dot blot hybridization on a to-be-detected sample, wherein the target DNA extraction is performed on the to-be-detected sample by adopting a NaI magnetic bead method; carrying out PCR (Polymerase Chain Reaction) amplification by taking the extracted DNA as a template and adopting a primer designed aiming at the Brucella BCSP-31 gene; carrying out reverse dot blot hybridization on a PCR amplification product on a hybridization membrane; wherein a detection probe designed aiming at a Brucella BCSP-31 gene is fixed on the hybridization membrane. The detection method disclosed by the invention can be fully automatically carried out by utilizing the micro-fluidic chip, is safe, rapid, timely and accurate, and the application of the technology disclosed by the invention can improve the diagnostic rate and diagnostic accuracy of Brucella, and plays an important role in preventing and treating Brucella diseases.