52results about How to "Rapid differential diagnosis" patented technology

The invention discloses a recombinase polymerase application (RPA) primer for rapidly detecting African swine fever virus (ASFV) nucleic acid, a preparation method of the RPA primer, and a kit, belonging to the technical field of biology. A pair of RPA primers, i.e., an upstream primer <210>2 and a downstream primer <210>3 which have high specificity and strong sensibility are screened out; an RPAdetection system for rapidly detecting the ASFV nucleic acid is further established by means of the primers. Compared with the common polymerase chain reaction (PCR) method, RPA-lateral flow assay (LFA) has the advantages that firstly, the RPA-LFA belongs to an isothermal amplification technology and is low in requirements for instruments and equipment, and reactions can be completed only by means of a constant temperature water bath kettle; secondly, the detection speed of the RPA-LFA is fast, and the reaction time is 40 minutes and is shorter than the conventional PCR reaction time; thirdly, the visualization of detection results can be realized. Due to the characteristics, the RPA-LFA method established by the invention can be used for rapid detection and differential diagnosis of ASFVin common laboratories of grassroots units.

The invention relates to a reaction system for detecting African swine fever virus nucleic acid. The reaction system is an SHERLOCK reaction system. The system comprises an RPA primer pair for amplifying a target nucleic acid fragment and / or crRNA, the crRNA is used for combining ssRNA transcribed by the amplified target nucleic acid fragment, the primer pair comprises primers with sequences shownas SEQ ID NO.1 and SEQ ID NO.2, the crRNA is synthesized from crDNA, and the sequence of the crDNA is shown as SEQ ID NO.3. The system further comprises Cas13a, a fluorescence labeling probe and thelike, the Cas13a is combined with the crRNA combined with the ssRNA so as to have RNA enzyme activity, the Cas13a with the RNA enzyme activity cuts the fluorescence labeling probe to generate fluorescence, and the generated fluorescence can be detected in real time. The invention also relates to a kit comprising the reaction system and related application of the RPA primer pair and / or the crRNA. The kit and a detection method are used for detecting the African swine fever virus nucleic acid, can realize constant-temperature detection at 37 DEG C, have short reaction time, high detection speed,and high sensitivity and the specificity.

The invention provides microRNA (micro ribonucleic acid) for identification of subtypes of a lung cancer. The miroRNA is chosen from the following sequence: a) a sequence shown as SEQ ID No. n (sequence identification number), or b) microRNA complementary to the sequence shown as SEQ ID No. n, wherein the n is 2, 3, 4 or 7. The invention further provides an application of the microRNA and a reagent kit based on the microRNA. The invention further provides a set formed by the microRNA, an application of the set and a reagent kit based on the set. The miroRNA has the advantages that the microRNA can form two combinations, namely SEQ ID No.2 and SEQ ID No.7, as well as SEQ ID No.3 and SEQ ID No.4, which can be used to differentially diagnose the small cell lung cancer, non-small cell lung cancer, adenocarcinoma lung cancer and squamous cell lung cancer respectively; the diagnosis is high in speed, low in cost and accurate and reliable in result; and a significant basis is provided for performing precise targeted therapy on a patient with the lung cancer.

The invention provides a fluorescent quantitative PCR detection reagent, detection kit and detection method for identifying and diagnosing infection and immunization of African swine fever, and belongs to the technical field of biology. Specific primers and probes are designed aiming at CD2v of the African swine fever and eGFP of a fluorescent protein. An ASFVP72 / CD2v dual fluorescent PCR detection method or ASFVP72 / CD2v / eGFP triple fluorescent PCR detection method formed through combination with a P72 gene can be used for detecting the infection of the African swine fever and identifying a wild strain of a African swine fever virus and a CD2v gene deletion type recombinant vaccine strain, and thus, infected animals and immunized animals can be differentiated. The fluorescent PCR detectionkit has the advantages of simplicity and convenience in operation, high sensitivity, strong specificity, good repeatability and the like, and an important monitoring measure is provided for epidemicsituation detection and disease prevention and control.

The invention discloses a DNA library for detecting idiopathic pulmonary fibrosis pathogenic genes with a targeted high-throughput semiconductor sequencing technique and an application of the DNA library. Specifically, a primer pool is designed according to 92 idiopathic pulmonary fibrosis pathogenic genes, super-multiplet PCR amplification is performed on sample genome DNA, an amplification product is sequenced with the high-throughput semiconductor sequencing technique, pathogenic mutations are searched, and a theoretical basis of genetics and molecular biology is provided for clinical diagnosis. The DNA library has the characteristics of being accurate, rapid, flexible and low in cost. Involved 92 gene detection regions can be used for detecting various types of idiopathic pulmonary fibrosis related to known genes and have great significance and clinical value in diagnosis and differential diagnosis of the idiopathic pulmonary fibrosis.

The invention relates to the technical field of immunochromatography, and particularly relates to a preparation method of an NSE test strip, and a test strip, a test card and an NSE test kit preparedaccording to the method. The preparation method of the NSE test strip comprises the following steps: arranging a sample pad, a bonding pad, a nitrocellulose membrane and an absorbent pad on a bottom plate in sequence, wherein an NSE antibody-fluorescent microsphere composite is fixed on the bonding pad, a detection line T and a quality control line C are arranged on the nitrocellulose membrane, the control line C is arranged at the end close to the absorbent pad, the detection line T is coated with an anti-NSE monoclonal antibody, and the control line C is coated with a goat anti-mouse IGg polyclonal antibody. The NSE test strip is used for detecting the NSE test speed without relying on large-scale medical equipment.

The invention relates to a targeted database creation kit for 43 pathogens. The pathogens comprise bacteria, viruses, fungi and protozoa. Targeted detection primer sequences of the pathogens are shownas SEQ ID NO. 1 to 86. According to the kit, targeted database creation can be performed on pathogen nucleic acids in an early infection stage for rapid diagnosis of the pathogens after next-generation sequencing, and directions are provided for subsequent pathogen isolation and identification, serologic detection, clinical diagnosis and symptomatic treatment.

The invention discloses primers for identifying an avian infectious bronchitis virus strain type, an RT-PCR detection kit, a method and application. Sequences of the primers are as follows: an H120 forward primer: 5'-ATTGCTTACGGTCCTCT-3', and an H120 reverse primer: 5'-CTGCTGGTTGACATCTT-3'; a QX forward primer: 5'-GTACAGGGTCTTGTCCTA-3', and a QX reverse primer: 5'-GTGTTGCTTAATTCACCT-3'; an LDT3 forward primer: 5'-CGCCACAGCAGGAAGAAT-3', and an LDT3 reverse primer: 5'-GTCCGTAGTTGGAATGAAGA-3'; a 4 / 91 forward primer: 5'-CCAGATAGGCGGTGTTAG-3', and a 4 / 91 reverse primer: 5'-TCGGCAATAGGAAAGTGT-3'. The primers, the RT-PCR detection kit, the method and the application have the beneficial effects that the kit provided by the invention separately amplifies four fragments of different sizes by only once gradient RT-PCR, and then, vaccine strains and popular wild strains of infectious bronchitis of chickens can be rapidly identified and diagnosed; the kit has the characteristics of high efficiency,low cost, high specificity, high sensitivity and the like; a convenient and quick detection method is provided for the differential diagnosis and molecular epidemiological analysis of avian infectious bronchitis viruses and prevention and control of the avian infectious bronchitis viruses.

The invention provides a primer group for identifying porcine circovirus types 1, 2 and 3 and application of the primer group. The sequences of the primer group are shown in SEQ ID NO.1 -4. On the basis of the genome structure characteristics of the three genotypes (PCV1, PCV2 and PCV3) of porcine circovirus, genome specific conserved areas of the three genotypes (PCV1, PCV2 and PCV3) of porcine circovirus are selected as upstream primer design areas, and downstream primers are designed separately in differential areas of genomes generated according to the three genotypes (PCV1, PCV2 and PCV3). The primer is simple in condition optimization, convenient and high in reaction speed, and a result is observed conveniently; an assembled kit can be used for rapid differential diagnosis of the three genotypes of porcine circovirus (PCV1, PCV2 and PCV3).

The invention relates to a protein chip for rapidly detecting pectoral ascites cancer cells and a preparation method thereof. The chip is provided with a solid matrix glass slide; and anti-tumor antigenic specific antibodies marked by various types of colloidal gold are combined and distributed at different positions on the surface of the glass slide in a dot matrix way. The preparation method comprises the following steps of: preparing the colloidal gold, the antibodies marked by the colloidal gold by using a trisodium citric acid reduction method; performing sample application on a prepared aldehyde group glass slide and the like. In a clinical practice, the cancer cells in pectoral ascites can be combined with the corresponding specific antibodies in the chip after pectoral ascites cast-off cell suspension and the chip are incubated together, and the combined cancer cells can be observed by a microscope so as to achieve the purpose of rapidly detecting and diagnosing cancerous pectoral ascites. The product has the characteristics of firm protein combination, good stability, difficult degradation, suitability for transportation and storage at room temperature, high throughput, time saving, low price and the like.

The invention discloses a dual fluorescent microsphere immunological detection method for pseudorabies virus gE and gB IgG antibodies. The detection method is based on a liquid protein chip technology-based carboxylated fluorescent microsphere group for detecting pseudorabies virus antibodies; the group comprises carboxylated fluorescent microspheres coupled with gE truncated proteins and gB truncated proteins, respectively; and the amino acid sequences of gE and gB truncated proteins are shown in SEQ ID NOs: 2 and 4, respectively. The dual fluorescent microsphere immunological detection method for simultaneously detecting PRV gE and gB IgG antibodies is good in reproducibility, high in sensitivity and good in specificity, and has no cross-reaction with other common porcine virus-positiveserums. The method can be used for the rapid differential diagnosis of pseudorabies virus wild-type infected pigs and vaccine-vaccinated pigs and the detection of protective antibodies, thereby providing an important method for the monitoring of swine virus diseases, having a great application value and being worthy of large-scale promotion.

The invention discloses a one tube PCR type kit for discriminating duck hepatitis virus (DHV-1A) and new I type duck hepatitis virus (DHV-1C), and the kit comprises a primer, a reagent for treating sample to be tested, a PCR reagent and an instruction, wherein, the primer is SEQ ID Nos:1-3. The kit of the present invention is suitable for identifying DHV-1A virus and DHV-1C virus in liver, spleen and kidney tissue samples, cloaca cotton swab, duck or chicken embryo allantoic liquid and cell culture sample of duck.

The invention relates to a virus detection method, in particular to a polymerase chain reaction (PCR) detection method universal for viruses. The method comprises the steps of sample treatment, nucleic acid extraction, amplification, electrophoresis observation, amplification product cloning, genome sequencing analysis and the like. Host genome infection, bacterial infection or exogenous virus infection are avoided by the PCR detection method universal for the viruses, and the method has better specificity and repeatability. The method is used for quickly identifying and diagnosing different viruses, the complexity of selecting and identifying a cause of disease caused by specific PCR one by one is overcome, and the method can be used for quickly detecting a clinically unknown virus infection sample during production and quickly and effectively detecting a new virus or a virus strain with high genetic variation.

The invention provides a reagent kit for detecting porcine delta coronaviruses and acute diarrhea syndrome coronaviruses porcine delta coronavirus. According to a primer and probe (SEQ ID NO: 1-6) fordouble real-time fluorescent quantitative PCR detection of the porcine delta coronaviruses and the porcine acute diarrhea syndrome coronaviruses, and a double real-time fluorescent quantitative PCR detection method established based on the primer and the probe, and rapid identification and detection of the two viruses in one reaction can be realized at the same time. The reagent kit can completedetection within 2 hours, has the advantages of quickness, specificity, sensitivity, high flux and the like, and can meet the requirements of quickly identifying and detecting the porcine delta coronaviruses and the porcine acute diarrhea syndrome coronaviruses on a large scale.

The invention discloses a kit, an RPA primer pair, a probe and a method for detecting CPV nucleic acid. The kit comprises an RPA reaction system, the RPA reaction system comprises an RPA primer probemixed solution, the RPA primer probe mixed solution comprises a primer pair and a probe, the nucleotide sequence of the primer pair is as shown in SEQ ID NO.1 and SEQ ID NO.2, and the probe is as shown in 5' SEQ ID NO.3+FAM-dt+THF+BHQ1-dt+SEQ ID NO.4+C3Spacer 3'. The kit is used for detecting canine parvovirus, has short reaction time and high detection speed, and has high sensitivity and specificity.

The invention provides a kit for detecting 4 / 91-type infectious bronchitis viruses, and belongs to the technical field of RT-PCR (reverse transcription-polymerase chain reaction) detection. The kit comprises a pair of specific primers, and nucleotide sequences of the specific primers are shown as SEQ ID NO.1 and SEQ ID NO.2 respectively. The invention further provides a method for detecting the 4 / 91-type infectious bronchitis viruses. The kit and detecting method has the advantages of accuracy in detection, high sensibility, high specificity, simplicity, convenience and rapidness and has good detection ability of clinical specimens.

The invention discloses a method for detecting mutation on the locus deltaF508 of a CFTR (Cystic Fibrosis Transmembrane Regulator) gene and an oligonucleotide, relating to a pair of specific amplification primers SEQ NO 1 and SEQ NO 2 and a pair of specific detection probes SEQ NO 3 and SEQ NO4. The method has the advantages of short detection period, high specificity, high accuracy, high sensitivity, low conditional dependency, low pollution risk and the like, and can be used for assisting in accurately and rapidly diagnosing cystic fibrosis and male infertility.

The invention relates to the field of immunocolloidal gold chromatography analysis, in particular to a kit for rapidly detecting natural infection antibodies of animal hydatid. The kit can specifically detect the natural infection antibodies of the hydatid, and has no cross reaction with the vaccine immune antibody. The kit comprises a colloidal gold test paper card and a sample diluent, wherein the colloidal gold test paper card comprises a closed box body and a reaction test strip tiling in the box body, the box body is respectively provided with an observation window and a sample adding hole connected to the reaction test strip; and the sample diluent is a mixed solution including 0.01M PH7.2 PBS and 1% tween-80. The invention provides a simple, rapid and scientific method for diagnosing echinococcosis infection, which can quickly detect whether a blood sample contains natural infection antibodies of animal hydatid.

The invention discloses a method and oligonucleotide for detecting FGFR3 gene G380R site mutation, and relates to a pair of specific amplification primers SEQ NO 1 and SEQ NO 2, and a pair of specific detection probes SEQ NO 3 and SEQ NO 4. By adopting the method and oligonucleotide, congenital achondroplasia can be rapidly diagnosed and identified, and the method and oligonucleotide have the advantages of being short in detection cycle, good in specificity, high in accuracy, high in sensitivity, less in condition dependence, low in pollution risk and the like.

The invention belongs to the technical field of animal pathogeny detection and particularly relates to primer and a reagent kit of differentiating virulent and attenuated Newcastle disease viruses, and a differential detection method. The differential detection method particularly comprises the steps of extracting RNA (ribonucleic acid) of strains to be detected, dissolving RNA with 30 microliter DECP (diethyl chlorophosphite) water, performing RT-PCR (reverse transcription-polymerase chain reaction), and conducting agarose gel electrophoresis test on a DNA (deoxyribonucleic acid) amplification result. The primer designed according to different nucleotide sequences of F gene cracking sites of the virulent and attenuated NDV (Newcastle disease virus) strains has good specificity and can accurately differentiate the virulent and attenuated Newcastle disease virus strains; the reagent kit is high in sensitivity, short in consumed time, and accurate and effective in detection, can detect to-be-detected samples, with nucleic acid not less than 100 ng / microliter, of the Newcastle disease viruses and facilitates quick clinical differential diagnosis.

The invention provides a portable Fourier infrared spectroscopy disease diagnosis system for a bone nonunion broken end. The system comprises a portable Fourier transform infrared spectrometer and anautomatic judging module adopting electric signal connection. The portable Fourier infrared spectroscopy disease diagnosis system for the bone nonunion broken end is used for carrying out disease diagnosis on a bone nonunion broken end sample. Based on the classification method of a support vector machine (SVM), the rapid differential diagnosis can be carried out on the bone nonunion type of patients during the surgery, and meanwhile, clinical guidance is provided for a clinician for accurately judging the osteotomy position of the bone nonunion broken end during the surgery. In addition, thedevice has the function of machine automatic learning, according to the increase of detected cases, the judging module can realize automatic updating and optimizing, and thus the accuracy rate in diagnosing the bone nonunion type and the efficiency in judging the osteotomy position during the surgery are improved.

The invention discloses an RPA (recombinase polymerase amplification) specific primer pair, a single-stranded DNA (deoxyribonucleic acid), a crRNA fragment, a CRISPR-Cas13a (clustered regularly interspaced short palindromic repeats) detection system and application thereof in preparation of ALV related diagnostic products and vaccines. RPA amplification can be carried out on target nucleic acid under the condition of constant temperature of 37 DEG C, the requirement for instruments and equipment is low, an amplified target fragment is transcribed into RNA, which is identified by specific crRNA and then is cut by Cas13a, the 'incidental cutting' activity of the Cas13a can cut RNA reporter molecules in the system, the RPA reaction time is about 30 minutes, the SHERLOCK reaction time is about 40 minutes, and the overall detection time is shorter than 2 h; According to the invention, when matched with the application of a lateral flow chromatography test strip, a detection result is visualized, and rapid detection and differential diagnosis of ALVA / B / J can be realized.

The invention discloses a degenerate primer group, a kit and a method for LAMP detection of a prawn white spot syndrome virus. The degenerate primer group comprises a primer OF, a primer OR, a primer IF, a primer IR and a primer LB, the primer OF: 5'-CTTAGACCATATKTAAGAGTG-3'; the primer OR:5'-GCAGATTCCAAACCAGGAA-3'; the primer IF: 5'-GAGCGGGCATAGAAGTGTAAAAMAT-TCTCTTCTTGGAATGATGAACG-3'; the primer IR: 5'-TCCAATATGAAGCAATGGGAGCTA-GGTTTAAGCCATGGTCTCT-3'; and the primer LB: 5'-GTGCTAATGGCAGCTGAA-3'. Specific amplification of the prawn white spot syndrome virus is completed by designing the degenerate primer group for LAMP detection of the prawn white spot syndrome virus, the specificity is good, the sensitivity is high, and an LAMP detection kit for the prawn white spot syndrome virus is designed on the basis. The LAMP detection kit can complete detection of various prawn white spot syndrome virus strains within 40 minutes, is high in detection speed and simple and convenient to operate, and can be widely applied to rapid differential diagnosis of various prawn white spot syndrome virus strains.

The invention provides a three-dimensional biological detection system based on fluorescent quantum dots, and a preparation method and application thereof, and belongs to the field of in-vitro diagnosis and detection. According to the three-dimensional biological detection system provided by the invention, the water-soluble fluorescent quantum dot is used as a fluorescent probe, a three-dimensional structure is combined as a detection substrate, a double-antibody sandwich method of 'coated antibody-antigen to be detected-fluorescent quantum dot labeled antibody' is utilized, and a simple, sensitive and stable novel method for detecting various markers is established. A detection signal can be amplified, the detection sensitivity can be improved, and meanwhile, the detection accuracy is ensured by good detection specificity, simplicity and convenience in operation and a stable fluorescent marker.

The invention discloses a recombinant expression vector for a lung fluke excretion and secretion protein, an engineering strain, a preparation method and an application thereof. The recombinant expression vector contains the encoding gene of the lung fluke excretion and secretion protein, the nucleotide sequence of the coding gene is shown as SEQ ID No.1, and the expression vector is pQE-80L. Theengineering strain contains the recombinant expression vector, and a host strain is bacillus coli. The method for preparing the lung fluke excretion and secretion protein through the recombinant expression vector comprises four steps of: constructing the engineering strain, culturing, inducting and expressing, and purifying an expression product. The lung fluke excretion and secretion protein canbe used for preparing agents or devices for detecting a paragonimiasis The recombinant expression vector and the engineering strain have the advantages of high expression amount and stable expression, and the lung fluke excretion and secretion protein prepared by the method has high protein yield and high immunoreactivity.

The invention discloses a recombinant expression vector for a lung fluke excretion and secretion protein, an engineering strain, a preparation method and an application thereof. The recombinant expression vector contains the encoding gene of the lung fluke excretion and secretion protein, the nucleotide sequence of the coding gene is shown as SEQ ID No.1, and the expression vector is pQE-80L. The engineering strain contains the recombinant expression vector, and a host strain is bacillus coli. The method for preparing the lung fluke excretion and secretion protein through the recombinant expression vector comprises four steps of: constructing the engineering strain, culturing, inducting and expressing, and purifying an expression product. The lung fluke excretion and secretion protein can be used for preparing agents or devices for detecting a paragonimiasis The recombinant expression vector and the engineering strain have the advantages of high expression amount and stable expression, and the lung fluke excretion and secretion protein prepared by the method has high protein yield and high immunoreactivity.

The invention provides a method for detecting avian paramyxoviruses, which detects RT-PCR of avian paramyxoviruses (APMV) by using specific primers. The method comprises the following steps of: performing extraction and reverse transcription of total sample RNA to obtain sample cDNA; amplifying target fragments by using the specific primers; performing gel electrophoretic analysis; and judging results. A pair of specific detection primers is designed according to conservative region sequence of NP genes of different serotype avian paramyxoviruses, the sequence is as shown in Seq ID NO:1 and Seq ID No:2, and target gene fragments of 387bp are amplified. The method has the characteristics of high specificity, high flexibility, high efficiency, low cost, suitability for quickly detecting the different serotype avian paramyxoviruses as well as analyzing and detecting a large number of samples and extremely important significance for quickly diagnosing the avian paramyxoviruses at early stage as well as analyzing molecular epidemiology, and simultaneously provides technical means for research on the molecular variation mechanism of the avian paramyxoviruses.

The invention discloses a kit for porcine reproductive and respiratory syndrome virus (PRRSV) gene-labeled vaccine strain ELISA differential diagnosis. The kit comprises NP49 polypeptide antigen and an amino acid sequence of the NP49 polypeptide antigen is shown in the formula of SEQ ID NO: 1. The kit for PRRSV gene-labeled vaccine strain ELISA differential diagnosis utilizes NP49 polypeptide as a coating antigen, has the characteristics of strong specificity, high sensitivity and good repeatability, can fast and accurately distinguish traditional PRRS vaccine- and labeled vaccine-immunized pig serum antibodies, can fast and accurately distinguish and diagnose the PRRSV gene-labeled vaccine strain and a natural infection strain, and has an important meaning for wide clinical application of the PRRSV gene-labeled vaccine strain.