22 results about "Housekeeping gene" patented technology

The application discloses a kit for diagnosing systemic lupus erythematosus, which is lanosterol lipid metabolite key enzyme CYP51A1, the expression amount of the transcription level of the lanosterol lipid metabolite key enzyme CYP51A1, and the cDNA sequence of the lanosterol lipid metabolite key enzyme CYP51A1 is shown as SEQ ID No. 1. The kit for diagnosing SLE provided by the application comprises CYP51A1 gene primers and primers of a housekeeping gene GAPDH, a standard product and amplification reagents. The application overcomes the defect that SLE gene diagnosis cannot be diagnosed. The application adopts a real-time fluorescent quantitative PCR method, has the characteristics of rapidness, sensitivity, accuracy and the like, the expression of CYP51A1 is detected through real-time fluorescent quantitative PCR, important basis and reference value can be provided for clinical diagnosis of diseases, thus being beneficial to the formulation of a treatment scheme of the disease, and having popularization and application value.

Provided is an internal reference gene for the real-time quantitative PCR (RT-qPCR) of Yersinia enterocolitica, including one of or a combination of some of glnS, nuoB, glmS, gyrB, dnaK, and thrS genes. Further provided is a primer set for amplifying the internal reference gene for the RT-qPCR of Yersinia enterocolitica. Given that stable housekeeping genes of Yersinia enterocolitica remain unclear, internal reference genes with relatively stable expression at different culture temperatures are selected by screening for Yersinia enterocolitica. Specific detection primers and real and reliable data provide a reference basis for selecting housekeeping genes of Yersinia enterocolitica; moreover, the use thereof alone or in combination, as an internal reference gene, improves the data accuracy, stability, and reliability, thereby solving the problem that usually only empirically selecting 16s rRNA as a single internal reference gene in conventional RT-qPCR results in unstable research results.

The invention belongs to the technical field of microbiological detection, and particularly relates to a Klebsiella pneumoniae MLST typing primer group and method based on a hospital localized microbiological mNGS platform. The primer group comprises primer pairs for amplifying an rpoB gene, an infB gene, a pgi gene, a phoE gene, a gapA gene, an mdh gene and a tonB gene, and corresponding nucleotide sequences are as shown in SEQ ID NO: 1-14. According to the technical characteristics of short sequencing read length (50 bp) and single-ended sequencing of a next-generation sequencing platform in a hospital, the primer group for Klebsiella pneumoniae MLST housekeeping gene amplification is redesigned, and amplicons are adaptive to the sequencing working principle of the existing next-generation sequencing platform in the hospital by optimizing the length and position of the amplicons. By utilizing the primer group, the kit or the method, the genetic relationship between hospital infection pathogenic bacteria can be quickly and accurately judged (within 24 hours), the timeliness and scientificity of infection prevention and control are remarkably improved, and the primer group, the kit or the method have a wide clinical application prospect.

The invention relates to a method and a kit for detecting SMA related gene copy number based on an NGS technology. The detection method comprises the following steps: extracting DNA of a sample to be detected and constructing an NGS library; designing a specific capture probe based on SMN1 / SMN2 differential sites a, b and c, and performing whole exon sequencing to obtain original sequencing data; performing data preprocessing on the sequencing original data; selecting a human control gene set as a housekeeper gene candidate set, and screening n genes close to SMN1 and SMN2 whole exon sequencing depth from the housekeeper gene candidate set as housekeeper genes k; standardizing the sequencing depth of each housekeeping gene k, and normalizing to obtain a theta value; the average value of all housekeeping genes theta is recorded as a correction factor value theta, and the copy states of the SMN1 and SMN2 genes in the DNA sample to be detected are corrected. The method has the beneficial effects that the quantitative accuracy of the copy number of the SMN1 and SMN2 genes is improved, and meanwhile, the detection efficiency is improved.

The invention provides a reference gene combination for real-time fluorescent quantitative PCR (Polymerase Chain Reaction) of indocalamus as well as a primer pair combination and application of the reference gene combination, and belongs to the technical field of plant molecular biology. In combination with indocalamus transcriptome data analysis and cross validation of four algorithms, reference genes, MD10B, PP2A, eIF1A, Ite23725, eIF4A, 60S and UBP1, suitable for indocalamus are systematically screened out, optimal reference gene combinations under different experimental conditions (drought, salt, waterlogging stress and different tissues) are refined, and the method is scientific and rigorous and has good application prospects. And a precise tool is provided for gene expression analysis of the indocalamus in different research directions. The screened reference genes show extremely high expression stability in specific stress and tissues after being combined, and the expression stability is far better than that of traditional housekeeping genes such as Actin and UBI. The specific primer has high amplification efficiency, and ensures the accuracy of qRT-PCR results.

The invention provides a quality control primer group and a quality control method for a human-derived sample, and belongs to the technical field of gene sequencing, and the quality control primer group comprises a human-derived DNA quality control primer designed based on conserved genes of GJB2MD, GJB3MD, IVSM and 12SD, a human-derived RNA quality control primer designed based on conserved and highly-expressed housekeeping genes of GAPDH, ACTB and PARP2, and a sex quality control primer designed based on genes of SRY142, DYS14134, YSR240 and YSR186 on a Y chromosome. According to the scheme provided by the invention, multi-dimensional and full-process monitoring on the human-derived sample can be realized, manual operation errors such as reagent failure, reagent addition error, sample confusion and cross contamination in the existing sequencing process are reduced, and the reliability of a detected result is effectively improved.

The invention discloses a bacillus cereus strain and application thereof, a phylogenetic tree is constructed through whole genome sequencing according to housekeeping genes, the phylogenetic tree is determined to belong to bacillus cereus (Bacillus cereus), and the preservation number is CGMCC (China General Microbiological Culture Collection Center) No.34800. The strain B2 obtained through separation can secrete indoleacetic acid (IAA), IAA serves as auxin and is beneficial for stimulating plant root development and promoting root growth, so that the water and nutrient absorption capacity of plants is enhanced, the adaptability of the plants in a stress environment is improved, meanwhile, the strain B2 can regulate and control sodium and potassium ion transport in pepper plants, and the yield of the pepper plants is increased. Specifically, the expression level of the key ion transporter gene in the pepper is up-regulated. The bacillus cereus strain B2 is an efficient microbial resource and can be used for developing a microbial inoculant or a biological fertilizer so as to promote growth of crops such as capsicum in salinized soil, improve the salt resistance of the crops and guarantee and improve the sustainable production capacity of capsicum in the salinized soil.

The application discloses a method, reagent and application for identifying or tracing citrus canker bacteria. The method comprises the following steps: obtaining seven gene sequences of the to-be-tested citrus canker bacteria, performing multi-locus sequence typing analysis on the seven gene sequences, determining alleles and numbering, concatenating the allele numbers of the seven genes to serve as a sequence type, comparing the sequence type with a reference strain sequence type, and obtaining tracing information of the to-be-tested strain; the reference strain is a citrus canker bacteria with known tracing information; the seven genes are XAC_RS07980, XAC_RS21550, copB, egl, fliL, hrpA and pliY. The seven housekeeping genes suitable for multi-locus sequence typing of the citrus canker bacteria are used for tracing detection, different geographical source strains can be distinguished at high resolution, and the method has important significance for strain regional research, evolution, transmission, disease prevalence and prevention and treatment. The method can be used for identifying and tracing the citrus canker bacteria.

The application discloses a primer composition and a reagent for fusion gene detection, and can detect human lung cancer fusion genes based on a multiplex PCR method. The primer composition can specifically amplify at least 50 fusion mutation forms of five fusion driver genes, such as ALK, NRG, NTRK1, RET and ROS1, in an RNA sample through sequences shown in SEQ ID NO 1-54, and can serve as a housekeeping gene for sample quality control. The reaction can be carried out in one tube or can be split into multiple tubes for carrying out. In the multiple tube reaction, a specific probe composition modified with a fluorescent group and having sequences shown in SEQ ID NO 55-70 can be added to form a qPCR reagent. When the qPCR reagent is used to detect a positive sample, the fusion gene in 0.04 ng of RNA can be reported. The process of the application is simple, efficient, easy to operate and low in cost, and has a good prospect.

The invention provides a primer for specifically detecting genome DNA residues in sisal hemp cDNA and a method and application thereof. According to the invention, a beta-ACTIN gene is found in housekeeping genes stably expressed in each tissue of sisal hemp, and a unique sequence difference exists in the beta-ACTIN gene of the sisal hemp by comparing a DNA sequence and a cDNA sequence of the genome. On the basis, a pair of specific primers is developed, cDNA and gDNA of the sisal hemp can be clearly distinguished in common agarose gel electrophoresis on the basis of the length difference of amplified fragments, and gDNA pollution residues can be accurately detected in sisal hemp of different sources and varieties. The detection primer has the advantages of being high in specificity, good in stability, high in universality, simple in detection method, economical, practical and the like, and an efficient and specific standardized quality control tool is provided for evaluating sisal hemp RNA extraction quality and guaranteeing reliability of subsequent molecular experiment data.

This invention discloses a method for constructing sequencing libraries based on blocking primers to remove human nucleic acids, and its application. The blocking primers include blocking primers for binding human repetitive sequences, blocking primers for binding human rRNA sequences, and blocking primers for binding human housekeeping gene sequences. These blocking primers can specifically bind to widely distributed scattered repetitive sequences, rRNA sequences, and housekeeping gene sequences in the human genome. The construction method includes the following steps: constructing a DNA library or an RNA library, or a co-constructed RNA and DNA library; using a library with added sequencing adapters as a template; and performing PCR amplification using adapter primers and blocking primers. The blocking primers can specifically bind to human nucleic acids through annealing and prevent modification by DNA polymerase extension, thus preventing the enrichment of human nucleic acid libraries through PCR amplification, while allowing normal amplification of pathogenic microorganism nucleic acids. This achieves the goal of reducing the proportion of human nucleic acids and provides guidance for pathogen detection.

The invention discloses a universal CRISPR / Cas12a (clustered regularly interspaced short palindromic repeats / CRISPR associated 12a) system, which can realize visual rapid detection of acinetobacter baumannii drug resistance genes. The system is combined with an asymmetric PCR technology to amplify one housekeeping gene and four drug-resistant genes of bacteria to generate a single-stranded DNA product, so that the amplification efficiency and the signal intensity are remarkably improved, and a high-specificity substrate is provided for subsequent reaction. The single-chain amplicon and the modular ring engineering hairpin are subjected to a strand displacement reaction, and Cas12a protein activation is accurately regulated and controlled; the activated Cas12a cuts the reporter molecule, and the product presents a visual visual signal through a lateral flow chromatography test strip. The strategy has prominent universality, crRNA does not need to be redesigned or an activating agent does not need to be added when five targets are detected, plug-and-play can be achieved only by replacing the hairpin response sequence, and the method can be conveniently expanded to detection of other bacteria, viruses and drug-resistant genes. The system is easy and convenient to operate, high in specificity and good in stability, provides a simplified and universal CRISPR diagnosis new scheme for clinical bacterial genotyping, and has important clinical application and popularization value.

The application discloses a blood tumor fusion gene micro residual lesion monitoring method, system and equipment, and technical scheme points thereof are as follows: high-throughput sequencing data of fusion gene information and all genetic information are acquired; the high-throughput sequencing data is compared to a reference genome without PCR deduplication, a first alignment result is obtained, real fusion gene sequences are screened from the first alignment result, a first copy number of the fusion gene is calculated according to the real fusion gene sequences; real housekeeping gene sequences are screened from the first alignment result, a second copy number of the housekeeping gene is calculated according to the real housekeeping gene sequences; and the first copy number is divided by the second copy number to obtain a micro residual lesion ratio of the fusion gene. The application can realize high-sensitivity and high-specificity monitoring of MRD.

The invention belongs to the technical field of agricultural microbial technology and plant disease biological control, and discloses bacillus velezensis SLW-15 antagonistic to agrobacterium tumefaciens and application of the bacillus velezensis SLW-15 antagonistic to agrobacterium tumefaciens, and the bacillus velezensis SLW-15 antagonistic to agrobacterium tumefaciens is separated from sakura tree rhizosphere soil suffering from crown goiter disease of cerasus campanulata grains in the Yunnan province, and is preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of CGMCC No.36629. The strain is accurately identified through morphology and multi-site housekeeping gene phylogenetic analysis. Experiments show that the strain SLW-15 and a fermentation product thereof have a remarkable antagonistic effect on agrobacterium tumefaciens. The invention optimizes the fermentation process (Landy culture medium, 32 DEG C, 220 rpm, 60 h) for producing antibacterial substances, and verifies that the produced antibacterial substances have good hot acid-base, enzyme activity and ultraviolet stability, and have an inhibition effect on various soil harmful bacteria. A pot experiment proves that the bacillus velezensis SLW-15 can be used for effectively preventing and treating the tomato crown goiter caused by agrobacterium tumefaciens.

The present application relates to the application of Paffia dura and its actin in removing cadmium in the environment; and a method for removing cadmium in the environment, comprising the following steps: exposing Paffia dura or microorganisms overexpressing actin from Paffia dura to an environment containing cadmium for incubation; removing Paffia dura from the environment. The present application finds that Paffia dura has high tolerance to cadmium stress and excellent cadmium enrichment capacity, and further finds that the main substance in Paffia dura cells combined with cadmium is actin. Therefore, Paffia dura and microorganisms overexpressing the actin can be used to treat cadmium-polluted environments to remove cadmium pollution. In addition, we also unexpectedly find that cadmium stress can up-regulate the expression of actin in Paffia dura, which is a new way to up-regulate the expression of housekeeping genes.

The invention provides a multi-site sequence typing primer and typing method of serratia marcescens, and belongs to the technical field of molecular detection of pathogenic microorganisms. The invention provides a specific primer group for multi-site sequence typing of serratia marcescens. The specific primer group comprises primer groups designed aiming at seven housekeeping genes: pitA, cyoB, glnA, ftsP, nadR, purR and potD. The invention also provides an MLST typing method of serratia marcescens. The MLST typing method comprises the following steps: extracting genome DNA of a strain to be detected; respectively carrying out PCR (Polymerase Chain Reaction) amplification by using the primer group; and sequencing the amplified product, and comparing the amplified product with a pre-established serratia marcescens MLST standard sequence database to obtain the sequence type of the strain. The invention provides a standardized MLST typing primer and method for serratia marcescens. The standardized MLST typing primer and method have the advantages of accurate typing, good repeatability, digital result, convenience in data sharing among laboratories and the like.

The invention relates to the technical field of molecular biology and in-vitro diagnosis, in particular to a multiple fluorescent PCR freeze-drying reagent and a kit for detecting a clostridium difficile housekeeping gene tpi, a toxin A gene and a toxin B gene. The freeze-drying reagent comprises reaction freeze-dried powder, and the reaction freeze-dried powder is prepared by freeze-drying a reaction solution prepared from components (a)-(f): (a) a first primer pair and a first probe for detecting a clostridium difficile tpi gene; (b) a second primer pair and a second probe for detecting the clostridium difficile tcdA gene; (c) a third primer pair and a third probe for detecting the clostridium difficile tcdB gene; (d) a fourth primer pair and a fourth probe for detecting an internal standard; (e) PCR reaction components, wherein the PCR reaction components comprise hot start Taq DNA polymerase, dNTPs (deoxyribonucleoside triphosphates) and a buffer system; and (f) a freeze-drying protective agent. The optimized system in the invention ensures that the triple targets of tpi (strain identification) and tcdA / tcdB (toxin typing) can still maintain high and consistent amplification efficiency after freeze-drying and redissolving, and the sensitivity can reach 5copies / reaction.

The invention discloses a swine salmonella primer composition, a kit and application, the primer composition comprises a wzy154 upstream primer, a wzy154 downstream primer, a donor probe and a receptor probe, and further comprises an ST typing primer capable of amplifying aroC, dnaN, purE, hemD, thrA, hisD and sucA. The primer composition can be used for identifying the serotype of the swine salmonella, and compared with a slide agglutination test, the primer composition is simple and convenient to operate, short in time consumption and low in requirements on experience and operation level of experimenters, and has remarkable cost and flux advantages. The ST typing primer is further integrated by optimizing the primer composition, and PCR independent amplification of seven housekeeping genes is optimized into five reactions, so that the typing efficiency is remarkably improved, and epidemiological monitoring and evolution research are facilitated.

This invention discloses a comprehensive identification method for *Peronobacter spp.* strain QH, belonging to the field of plant pathogen identification technology. The method includes the following steps: obtaining a sample of the *Peronobacter spp.* strain to be identified; extracting morphological characteristics of hyphae, sporangia, and conidiophores; extracting key morphological parameters and generating a morphological feature vector; amplifying its housekeeping gene fragment using polymerase chain reaction and sequencing to obtain the nucleotide sequence; calculating sequence similarity and generating a molecular feature vector; performing comprehensive matching and judgment, and outputting the identification result; generating an identification report, and combining it with strain information in a database for source tracing analysis and risk warning. This invention solves the problems of traditional identification methods relying on single indicators, strong subjectivity, and difficulty in distinguishing intraspecific genetic variations by multimodal fusion of morphological quantitative characteristics and molecular biological marker characteristics. It achieves accurate, objective, and automated identification of *QH* strains, providing data support for disease source tracing and control decisions.

This invention discloses a dual-luciferase reporter cell line for detecting CRISPR-Cas protein cleavage activity and its applications. The reporter cell line of this invention expresses the nanoluciferase gene Nluc with the housekeeping gene ACTG1, avoiding the influence of intracellular environmental changes on the activity of the nanoluciferase Nluc. Changes in Nluc activity reflect the cleavage activity of the CRISPR-Cas protein, overcoming the shortcomings of existing GFPs, which have a long half-life and cannot quickly respond to double-strand breaks at gene sites. Furthermore, the reporter cell line of this invention can also be used to screen CRISPR-Cas protein inhibitors. This invention has used the constructed reporter cell line to screen small molecule compounds that can inhibit SpCas9 cleavage activity intracellularly. The screening process is simple, convenient, and provides stable signals.

The application relates to a primer group, a kit and a method for detecting a plurality of disease-related gene mutation types. The application can simultaneously detect genetic deafness, thalassemia and spinal muscular atrophy disease-related gene mutation types, only needs one tube, and can simultaneously detect clinically common known pathogenic SNPs, INDELs and copy number variations in one experiment, thereby significantly improving the detection efficiency of the three diseases and the standardization degree of clinical detection. The primer group comprises: 1) a primer for detecting genetic deafness gene mutation types, as shown in SEQ ID NO: 1-30; 2) a primer for detecting thalassemia gene mutation types, as shown in SEQ ID NO: 31-340; 3) a primer for detecting spinal muscular atrophy gene mutation types, as shown in SEQ ID NO: 341-350; and 4) a primer for detecting a beta-actin housekeeping gene, as shown in SEQ ID NO: 351-352.

The embodiment of the invention relates to a drug resistance gene detection method and device, electronic equipment and a storage medium. The method comprises the following steps: acquiring first sequencing data corresponding to a to-be-detected sample and second sequencing data corresponding to a negative control sample; obtaining a noise baseline value according to the coverage degree of the second sequencing data in the target area; calculating a stability coefficient corresponding to the first sequencing data according to the original abundance corresponding to the at least one non-drug-resistant housekeeping gene; correcting the original abundance corresponding to the target gene in the first sequencing data based on the noise baseline value and the stability coefficient to obtain the target abundance corresponding to the target gene; according to the first sequencing data and the target abundance corresponding to the target gene, drug resistance detection is conducted on the target gene, a detection result corresponding to the target gene is obtained, and the detection result is used for indicating whether the target gene is a drug resistance gene or not. According to the drug-resistant gene detection method and device, the electronic equipment and the storage medium, the detection accuracy of the drug-resistant gene can be improved.