43results about How to "Primer specificity is high" patented technology

The invention discloses a universal RT-PCR (Reverse Transcription Polymerase Chain Reaction) detection primer and a detection method for avian pneumovirus (APV), belonging to the technical field of detection for avian viruses.. The universal RT-PCR detection primer has nucleotide sequences shown as SEQ ID NO:1 to SEQ ID NO:2. The detection method is as follows: the universal RT-PCR detection primer for avian pneumovirus and the RNA (Ribonucleic Acid) of a sample to be detected are subjected to RT-PCR, and the presence of avian pneumovirus in the sample to be detected is confirmed if the PCR product fragment is 319bp o 320bp. The method disclosed by the invention is high in primer specificity, small in fragment and high in sensitivity, can finish the entire PCR process within 2 hours and can also specifically detect the presence of various subtype APVs, thus greatly reducing the erroneous diagnosis for APV infection.

The invention provides a method and a system for realizing Y-STR typing on the male individual by adopting 26 Y-STR genetic loci. The method comprises the following steps: acquiring DNA of the male individual, and acquiring the genotypes of the 26 Y-STR genetic loci of the DNA, wherein the 26 Y-STR genetic loci are as follows: DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS448, DYS456, DYS458, DYS635, GATA H4, DYS438, DYS439, DYS460, DYS447, DYS527a/b, DYS617, DYS522, DYS444, DYS508 and DYS533; and acquiring the Y-STR typing result of the male individual according to the genotypes of the 26 genetic loci of the male individual. With the typing result, the operations including family checking, paternity identification, detection for the male element in male and female mixed samples, and the detection for the male element in a plurality of male mixed samples can be realized on the male individual.

The invention relates to the technical field of security detection of agricultural and sideline products, in particular to a primer for simultaneously detecting five food-borne pathogenic bacteria through a liquid chip and a detection method. The detection method comprises steps as follows: a specific primer and a gene probe for five foodborne pathogenic bacteria are designed; DNA templates of the detected five foodborne pathogenic bacteria are prepared; PCR (polymerase chain reaction) amplification is performed on the five foodborne pathogenic bacteria; coupling is performed on the gene probe for the five foodborne pathogenic bacteria and microspheres, and a microsphere probe is obtained and verified; an obtained PCR amplification product and the microsphere probe have a hybridization reaction; a fluorescence signal of a hybridized product is detected, and quantitative analysis is performed on the five foodborne pathogenic bacteria. A method capable of simultaneously detecting five common foodborne pathogenic bacteria is successfully established, the variable coefficient is within 2%, the specificity is 100%, the sensitivity can reach 100 CFU/mL, and the method is equivalent to a fluorescent PCR technology and can be applied to high-throughput rapid detection of clinic and environment specimens of foodborne diseases.

The invention discloses a method and a kit for detecting mutation of an EGFR gene. The method comprises the steps of bonding a specific primer with a target template DNA sequence, amplifying a to-be-tested sample target region by virtue of a universal primer, purifying a library by virtue of magnetic beads, carrying out high-throughput sequencing on the obtained library, and analyzing the mutation of the EGFR gene. The kit comprises the primers, a PCR mixing reagent, a positive control and the magnetic beads. By utilizing the method and the kit, the experimental steps can be simplified, and the detection sensitivity is improved. The method and the kit are applicable to the rapid detection of the mutation of the EGFR gene of clinic tumor patients and are helpful for guiding clinic personalized medication.

The invention provides method and system for Y-STR typing of an individual man. The method comprises the following steps: acquiring the DNAs of the individual man, wherein acquired DNAs comprise genotypes of 25 gene loci including 24 Y-STR gene loci and a gender determination locus Amelogenin, and the 24 Y-STR gene loci are DYS460, DYS389I/II, DYS390, DYS392, DYS458, DYS437, DYS385a/b, GATA_H4, DYS522, DYS456, DYS391, DYS447, DYS438, DYS448, DYS393, DYS635, DYS439, DYS19, DYS444, DYS527a/b, DYS617; and acquiring the Y-STR typing results of the individual man according to the 25 gene loci of the individual man. The Y-STR typing results can be used for family checking, male paternity testing, detection of male components in a hybrid male and female sample, detection of male components in a mixed sample of a plurality of males and the like for the individual man.

The invention provides an SNP marker related to a pepper tomato spotted wilt virus disease resistance gene as well as specific primers and application of the SNP marker. A nucleotide sequence of the SNP marker related to the pepper and tomato spotted wilt virus disease resistance gene is as shown in SEQ ID NO.1, and the SNP marker is a 21st basic group of the sequence as shown in SEQ ID NO.1. Theinvention also provides the specific primers and a corresponding test kit for identifying the SNP marker, and provides an identification method for pepper resisting the tomato spotted wilt virus disease by using the specific primers or the test kit. By adopting the SNP molecular marker disclosed by the invention, the pepper TSWV (Tomato Spotted Wilt Virus) disease resistance gene can be quickly and accurately judged only by carrying out DNA detection on the pepper plant in a seedling stage without artificial inoculation identification of pepper TSWV disease resistance, and large-batch detection can be carried out on samples, so that the breeding efficiency is improved, and the identification time is shortened.

The invention discloses a DNA examination reagent for sample examination, and a special primer thereof. A DNA examination primer group provided by the invention is composed of a primer pair group used for amplifying 21 Y-STR sites, a primer pair for amplifying a sex site Amelogenin, and a primer pair for amplifying an autosome SRT site. Experiments prove that the DNA examination reagent can simultaneously examine the 21 Y-STR sites and the autosome SRT site, and the primer specificity is high, so complete SRT typing can be obtained, peak patterns are sharp and intelligible, the balance is good, no Pull-up peaks or stutter bands appear, no non-specific artificial products appear, and legal medical experts' DNA large scale sample examination requirements are completely met.

The invention discloses a detection method for EV71 virus in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect EV71 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at EV71 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect EV71 virus in the environmental water body accurately.

The invention discloses a detection method for CVA16 in an environmental water body, wherein a semi-nested RT-PCR (Reverse Transcription-Polymerase Chain Reaction) and real-time fluorescence quantification RT-PCR reaction system is established to detect CVA16 virus in the environmental water body by using a recombinant plasmid as a reference product, and the recombinant plasmid is prepared by using a primer and a probe aiming at CVA16 virus designed according to a genetic conservative fragment of enterovirus belonging to a VP1 serotype area. The method has good specificity and high sensitivity and precision, and can qualitatively and quantitatively detect CVA16 virus in the environmental water body accurately.

The invention discloses a quick amplifying reagent kit and amplifying method of a CYP2C19 gene. The reagent kit comprises 3 pairs of amplifying primers, Taq enzymes, dNTPs and a buffered solution, wherein the amplifying primers are used for amplifying at least two SNP sites in a CYP2C19*2 type, a CYP2C19*3 type and a CYP2C19*17 type, and each pair of primers corresponds to upstream and upstream regions of one of SNP sites. A multiplex-PCR amplifying method is used, a plurality of important SNP sites of the CYP2C19 are amplified at the same time, the combination of sites to be amplified can beflexibly selected according to actual demands, and the effect that one reagent kit has many functions can be achieved; the primers in the reagent kit are high in specificity and low in mispairing rate, the length and the Tm value of the primers are reasonable to design, and amplified products are convenient to detect and separate. Multiple purpose products are amplified at a time, so that the amplifying efficiency can be greatly improved, and the amplifying cost can be reduced.

The invention belongs to the technical field of biology, and provides a special primer for simultaneously and accurately detecting SNP loci of related genes of 'slap-induced deafness' and ' injection-induced deafness'. The special primer comprises a primer group 1 and a primer group 2, wherein the primer group 1 is a specific PCR primer composed of 13 pairs of primers, and the primer group 2 is asingle-base extension primer composed of 13 primers. The invention further provides a preparation for simultaneously and accurately detecting the SNP loci of the related genes of 'slap-induced deafness' and 'injection-induced deafness', and a method for simultaneously and accurately detecting the SNP loci of the related genes of 'slap-induced deafness' and 'injection-induced deafness'. By adoptingthe special primer provided by the invention, accurate prediction of high-risk groups of 'slap-induced deafness' and 'injection-induced deafness' can be efficiently completed simultaneously through one-time detection, the detection time is greatly shortened, the cost is saved, and the special primer has very high application value for preventing 'slap-induced deafness' and 'injection-induced deafness' and 'deafness-induced dumbness' caused by 'slap-induced deafness' and 'injection-induced deafness'.

The invention provides a molecular marker of a rice brown planthopper-resistant gene BPH39 and an application thereof. A local high-density physical map covering the BPH39 is constructed by constructing a near-isogenic line of the BPH39 and analyzing a recombinant exchange single plant of a high-generation backcross population, and the gene is finely positioned in an interval of 30kb. The two molecular markers ZL1607 and ZL2137 in the interval are closest to the BPH39, and the reliability of a single plant for screening a target gene by utilizing the two markers is the highest. Besides, othermolecular markers, I1540, I32-4 and ZL6061 which are closely linked with the gene can be used for screening single plants of the target gene, so that the selection efficiency of brown planthopper-resistant rice varieties or strains can be greatly improved, and the breeding age limit is shortened.

The invention discloses a method for fast identifying the generation of an EC (ethyl carbamate) precursor of citrulline from lactic acid bacteria, and belongs to the technical field of microbiological detection. By the method, whether a lactic acid bacterium strain has an arginine deiminase path or not and whether citrulline can be formed by using arginine or not can be judged, so that whether the lactic acid bacterium strain in the fermented food and beverage has the possibility to generate a cancerogen of EC or not can be determined. The method disclosed by the invention is fast, simple, convenient and accurate; the degenerate primer specificity is high; the universality is high; the lactic acid bacteria of different species from different food can be detected; good food safety detection prospects are realized.

The invention relates to the technical field of rice breeding and molecular biology, in particular to a major QTL for regulating and controlling the brown rice rate of rice, a molecular marker and application. The invention discloses a major QTL for regulating and controlling the brown rice rate of rice. The major QTL is located on a chromosome 10 of the rice and is named as QBRR-1, the genetic distance is 70.1-81.73 cM, and the physical distance is 17570591-19066686 bp. The invention also provides the molecular marker of the major QTL for regulating and controlling the brown rice rate of the rice. According to the major QTL, the rice with high brown rice rate is bred through the molecular marker.

The invention relates to a method and kit for identifying genuine medicinal materials herba cynomorii different in growing area. The method comprises the steps that firstly, genome DNA of a to-be-detected sample serves as a template, and the segment containing the sequence shown in SEQ ID NO.1 is amplified; secondly, an amplified product is sequenced, after splicing, a primer area is deleted so as to obtain a complete amplified segment containing the sequence shown in SEQ ID NO.1, and basic groups at the 188th position and the 304th or 305th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 are detected; if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is T, or the basic group at the 304th position is A, or the basic group at the 305th position is C, the herba cynomorii is identified as Tacheng herba cynomorii, and if the basic group at the 188th position starting from the 5' terminal in the sequence shown in SEQ ID NO.1 is C, or the basic group at the 304th position is T, or the basic group at the 305th position is T, the herba cynomorii is identified as herba cynomorii from other growing areas. The method is wider in adaptability, high in primer specificity, high in amplification efficiency and capable of detecting any samples of herba cynomorii from which DNA can be extracted.

The invention belongs to the technical field of biology, and provides a special primer for detecting SNP sites of invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation.The special primer comprises a first primer group and a second primer group; the first primer group is a specific PCR primer consisting of three pairs of primer groups; and the second primer group isa single base extension primer consisting of three primers. The invention also provides a preparation of the SNP sites for detecting the invasive aspergillosis risk after allogeneic hematopoietic stemcell transplantation, and an SNP sites information method for detecting the invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation. By adopting the special primer for detecting the SNP sites of the invasive aspergillosis risk after allogeneic hematopoietic stem cell transplantation provided, the correlation between the gene polymorphism of the three SNP sites relatedto the attack risk of invasive aspergillosis after allogeneic hematopoietic stem cell transplantation and the occurrence risk of invasive aspergillosis after allogeneic hematopoietic stem cell transplantation can be efficiently and simultaneously completed through one-time detection, so that the detection time is greatly shortened, and the cost is saved.