39 results about "Housekeeping gene" patented technology

The invention relates to the technical field of genetic engineering, and particularly provides a multi-linked quality control product for quality control of a novel coronavirus nucleic acid detection laboratory and a preparation method of the multi-linked quality control product. The multi-linked quality control product is a multi-component composite system and is formed by mixing three substances, namely pseudovirus, target gene RNA (Ribonucleic Acid) molecules and internal standard plasmid DNA (Deoxyribose Nucleic Acid). Wherein the pseudovirus is pseudovirus particles of novel coronavirus ORF1ab gene RNA (Ribonucleic Acid) wrapped by a vesicular stomatitis virus vector and is used for monitoring virus nucleic acid extraction; the target gene RNA molecule is a novel coronavirus N gene RNA molecule and can monitor the reverse transcriptase activity; the internal standard plasmid is plasmid DNA of a human housekeeping gene RPP30, and whether inhibition or failure exists in an amplification link is monitored. According to the method, the whole process of nucleic acid extraction, reverse transcription and amplification is synchronously and hierarchically monitored, the monitoring blind area of a traditional single-component quality control product is solved, and the accuracy and efficiency of laboratory detection quality control are remarkably improved.

The invention discloses a blood tumor fusion gene minimal residual focus monitoring method, system and equipment, and the key points of the technical scheme are as follows: obtaining fusion gene information and high-throughput sequencing data of all genetic information; under the condition that the high-throughput sequencing data is subjected to PCR without duplicate removal, comparing the high-throughput sequencing data to a reference genome to obtain a first comparison result, screening out a real fusion gene sequence from the first comparison result, and calculating to obtain a first copy number of a fusion gene according to the real fusion gene sequence; a real housekeeping gene sequence is screened out from the first comparison result, and a second copy number of housekeeping genes is calculated according to the real housekeeping gene sequence; dividing the first copy number by the second copy number to obtain the minimal residual focus ratio of the fusion gene. According to the invention, high-sensitivity and high-specificity monitoring of MRD can be realized.

The invention provides a multiplex qPCR (quantitative polymerase chain reaction) detection primer and probe composition for detecting four psychrophilic bacteria and enzymes thereof and application, and belongs to the technical field of biological detection. The invention develops two quadruple qPCR methods for housekeeping genes and enzyme coding genes of pseudomonas, acinetobacter, bacillus cereus and stenotrophomonas maltophilia, the method has good specificity and versatility, the primer and the probe both show a good linear relationship between a Ct value and a colony count in single and multiple qPCR, and the primer and the probe can be used for detecting the colony count of the pseudomonas, the acinetobacter, the bacillus cereus and the stenotrophomonas maltophilia and the enzyme coding genes of the pseudomonas, the acinetobacter, the bacillus cereus and the stenotrophomonas maltophilia. The detection limit of the housekeeping gene is 101 CFU / mL, and the detection limit of the enzyme coding gene is 102 CFU / mL. The detection method can be used for evaluating the content of psychrophilic bacteria in milk and dairy products, the detection time is shortened to 2.5 h from 25 h, and the method has the advantages of being efficient, rapid and easy to operate.

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.

The invention provides an organ transplantation susceptible virus nucleic acid detection primer probe composition and a kit, and belongs to the technical field of biological detection. The primer probe composition comprises an upstream primer, a downstream primer and a fluorescent probe sequence, wherein the upstream primer and the downstream primer are designed for human cytomegalovirus, JC virus, BK polyoma virus, EB virus, human herpes virus type 6, human herpes virus type 8, human parvovirus B19, torque teno virus, new coronavirus and hepatitis E virus respectively; an upstream primer, a downstream primer and a fluorescent probe sequence of an internal standard are designed aiming at a human housekeeping gene RNaseP. The kit can effectively distinguish 10 viruses, ensures high accuracy, specificity and sensitivity, can complete detection in a short time to obtain a result, and can be transported and stored at normal temperature.

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 present application relates to the technical field of genetic engineering, and specifically provides a multi-link quality control product for novel coronavirus nucleic acid detection laboratory quality control and a preparation method thereof. The multi-link quality control product is a multi-component composite system, which is formed by mixing three substances of a pseudo virus, a target gene RNA molecule and an internal standard plasmid DNA. The pseudo virus is a pseudo virus particle of a vesicular stomatitis virus vector wrapping a novel coronavirus ORF1ab gene RNA, which is used for monitoring virus nucleic acid extraction; the target gene RNA molecule is a novel coronavirus N gene RNA molecule, which can monitor reverse transcriptase activity; and the internal standard plasmid is a plasmid DNA of a human source housekeeping gene RPP30, which monitors whether there is inhibition or failure in the amplification link. The present application realizes synchronous layered monitoring of the whole process of nucleic acid extraction, reverse transcription and amplification, solves the monitoring blind area of traditional single-component quality control products, and significantly improves the accuracy and efficiency of laboratory detection quality control.

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 discloses a primer combination for body fluid detection based on an RT-LAMP technology, a composite system and application, and belongs to the technical field of forensic medicine identification. The kit comprises a housekeeping gene, a specific mRNA (messenger ribonucleic acid) site and a primer combination for amplifying a corresponding site, each group of primers in the primer combination comprises two inner primers, two outer primers and two loop primers, and the specific sequences of the primers are as shown in SEQ ID NO.1-60. The LAMP primer is designed aiming at the body fluid specific mRNA site, the expression level of mRNA is detected, and then a body fluid sample is recognized. The method is simple to operate, high in specificity, strong in tolerance, strong in recognition capability and short in time consumption, can obtain a body fluid type result within 30 minutes, and can be applied to the field of forensic body fluid identification and the like.

The invention discloses a method and device for extracting the contour of an object, electronic equipment and a medium, and the method comprises the steps: dividing the object into at least one sub-graph, and obtaining an expression quantity graph set of the sub-graph for each sub-graph; for each expression quantity diagram in the expression quantity diagram set, calculating the distance between any two coordinate points in the expression quantity diagram, and deriving a divergence value of a gene associated with the expression quantity diagram based on the distance; generating a trend chart of the divergence values of the genes on the subgraph according to the derived divergence values to obtain mutation points in the trend chart, and generating a first subset based on the mutation points; fitting the distribution of the divergence values of the genes on the subgraph, and taking the genes in a preset interval in the fitted distribution as a second subset; generating a third subset and a fourth subset based on a housekeeping gene and a marker gene of the subject, respectively; generating a candidate set based on the first subset, the second subset, the third subset, and the fourth subset; and extracting the contour of the object according to the candidate set.

The invention discloses a freeze-dried microsphere for detecting novel coronavirus nucleic acid and a preparation method thereof, and the freeze-dried microsphere comprises a first upstream primer, a first downstream primer and a first probe which are designed in a novel coronavirus specific target N gene region, the first upstream primer, the second downstream primer and the second probe are designed in a novel coronavirus specific target ORF1ab gene region, the third upstream primer, the third downstream primer and the third probe are designed in a novel coronavirus specific target S gene region, and the fourth upstream primer, the fourth downstream primer and the fourth probe of a specific human housekeeping gene RNase P are designed in a novel coronavirus specific target S gene region. The kit comprises the following components: a reaction buffer solution, a UNG enzyme, a reverse transcriptase, a DNA polymerase, a freeze-drying protective agent and DEPC water. The freeze-dried microspheres can be stored at room temperature for a long time, the reagent performance is good, tedious operation in the liquid preparation process of an existing liquid type novel coronavirus nucleic acid detection reagent is simplified, experimental errors are reduced, and pollution is avoided.

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 housekeeping genes of pituitary, liver, ovary and liver fat storage cells of scatophagus argus and RT-qPCR (real-time quantitative polymerase chain reaction) amplification primers, and belongs to the technical field of genes. 17 pituitary candidate genes, 4 ovary candidate genes, 4 liver tissue candidate genes and 4 liver fat storage cell candidate genes are amplified through an RT-qPCR method, stability analysis is carried out through four analysis methods of GeNorm, NormFinder, Ct and BestKeeper, finally, sequencing is carried out through RefFinder, and results show that gnb2l1 and rpl4 in pituitary, eef1a and hprt1 of ovary, actb and hprt1 of liver tissue, rpl1 and rpl2 in liver tissue, rpl3 and rpl4 in liver tissue, rpl3 and rpl4 in liver tissue, rpl3 and rpl4 in liver tissue, rpl3 and rpl4 in liver tissue, rpl4 and rpl4 in liver tissue b2m and actb of the liver fat storage cells can be used as housekeeping genes of corresponding tissues and cells of scatophagus argus.

The invention belongs to the technical field of microbiological detection, and particularly relates to a pseudomonas aeruginosa MLST typing primer group and method based on a hospital localized microbiological mNGS platform. The primer group comprises a primer pair for amplifying an aroE gene, a trpE gene, an acsA gene, a ppsA gene, a nuoD gene, a guaA gene and a mutL gene, and corresponding nucleotide sequences of the primer pair 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 amplifying the MLST housekeeping gene of the pseudomonas aeruginosa is redesigned, and the length and the position of an amplicon are optimized, so that the amplicon adapts to the sequencing working principle of the existing next-generation sequencing platform in the hospital. 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 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 microbiological detection, and particularly relates to a staphylococcus aureus MLST typing primer group and method based on a hospital localized microbiological mNGS platform. The primer group comprises primer pairs for amplifying a yqiL gene, a pta gene, a tpi gene, a gmk gene, a glpF gene, an aroE gene and an arcC 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 invention redesigns a staphylococcus aureus MLST housekeeping gene amplification primer group, and by optimizing the length and position of amplicons, the amplicons are adaptive to the sequencing working principle of the existing next-generation sequencing platform in the hospital. 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 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 invention relates to a diagnostic kit for preeclampsia, a biomarker of the preeclampsia is Syncytin-1, a detection primer group is designed according to a gene sequence of the Syncytin-1 and a human housekeeping gene beta-actin sequence, a forward primer of the Syncytin-1 is as shown in SEQ ID NO.1, a reverse primer of the Syncytin-1 is as shown in SEQ ID NO.2, a reverse primer of the Syncytin-1 is as shown in SEQ ID NO.3, and a reverse primer of the Syncytin-1 is as shown in SEQ ID NO.4. A reverse primer of the Syncytin-1 is as shown in SEQ ID NO. 2; the fluorescent probe of the Syncytin-1 is as shown in SEQ ID NO. 3; a forward primer of beta-actin is as shown in SEQ ID NO. 4; a reverse primer of the beta-actin is as shown in SEQ ID NO. 5; and a fluorescent probe of beta-actin is as shown in SEQ ID NO. 6. According to the invention, a molecular biology method is innovatively adopted to diagnose preeclampsia of the pregnant woman, so that a basis is provided for early intervention and customized treatment, and early risk assessment and intervention in an early pregnancy period become possible.

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 belongs to the technical field of microbiological detection, and particularly relates to an escherichia coli MLST typing primer group and method based on a hospital localized microbiological mNGS platform. The primer group comprises a primer pair for amplifying a dinB gene, an icdA gene, a pabB gene, a polB gene, a putP gene, a trpB gene, a trpA gene and an uriA gene, and corresponding nucleotide sequences are as shown in SEQ ID NO: 1-16. The primer group comprises a primer pair for amplifying the dinB gene, the icdA gene, the pabB gene, the polB gene, the putP gene, the trpB gene, the trpA gene and the uriA gene. 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, an amplification primer group for the escherichia coli MLST housekeeping gene is redesigned, and amplicons are adaptive to the sequencing working principle of the existing next-generation sequencing platform in the hospital by optimizing the lengths and positions 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 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 discloses a primer design method for housekeeping genes, which belongs to the field of primer design methods, and comprises the following steps: acquiring transcriptome expression quantity data of a target species, and downloading an expression gene sequence file of the target species at the same time; processing transcriptome expression quantity data according to a preset screening rule to obtain a proper gene ID number which can be used as a housekeeping gene; according to the gene ID number of the house-keeping gene, searching a corresponding gene sequence in an expression gene sequence file of the target species to obtain a gene sequence of the house-keeping gene; and designing a primer based on the gene sequence of the housekeeping gene, and storing the designed primer sequence to an output file. According to the method, the housekeeping genes with stable expression and high expression are screened through transcriptome data, so that the steps of manual screening and verification are reduced; gene sequence information is processed on a large scale by means of automatic primer design software, a primer design scheme meeting experimental requirements can be rapidly generated, the accuracy of a design result is ensured, and the process is efficient.