50 results about "Gene conservation" patented technology

The invention belongs to the field of in-vitro diagnosis and detection, and particularly relates to a primer pair, a probe, a kit and a detection method for norovirus detection. The norovirus detection primer pair provided by the invention comprises a primer pair for detecting a norovirus GI type, a primer pair for detecting a norovirus GII type and a primer pair for detecting a norovirus GIV type, and specific primers and probes are designed on the basis of a norovirus GI / GII / GIV type gene conserved region; and a plurality of pairs of primers or probes are introduced, and degenerate primers and degenerate probes are introduced, so that the existing norovirus subtypes are better matched, the detection coverage rate is increased, and the omission ratio is reduced.

The invention provides a loop-mediated amplification primer group, a kit and a detection method for detecting influenza A virus, influenza B virus and respiratory syncytial virus, and relates to the technical field of biotechnology. Specific loop-mediated amplification primers are respectively designed aiming at M1 protein genes of influenza A viruses, NEP genes of influenza B viruses, F gene conserved segments of respiratory syncytial viruses and human RnaseP genes, and each set of primers comprises two outer primers (OF / OB), two inner primers (FIP / BIP) and at least one loop primer (LF / LB). Compared with a traditional detection method, the detection time is greatly shortened, a basis can be provided for clinical diagnosis more quickly, treatment measures can be taken in time, complex nucleic acid extraction and purification steps are not needed, the swab sample can be detected after being simply treated, the operation difficulty and the requirement for professional skills of operators are reduced, and the detection efficiency is improved. And popularization and application in all levels of medical institutions are facilitated.

The technology disclosed relates to detecting gene conservation and expression preservation. In particular, the technology disclosed relates to detecting gene conservation and epigenetic signals for a reference genetic sequence in comparison to a variant of the reference genetic sequence at base resolution through the generation of a plurality of alternative representations of the sequence in chromatin form which may represent evolutionary conservation, transcription initiation, or epigenetic signals, mapping the plurality of alternative chromatin sequences to a gene expression alterability classifier to generate a gene expression class prediction for the variant, and mapping the alternative chromatin sequence to a pathogenicity predictor to detect pathogenicity of variants.

The invention discloses a method for detecting a beta-lactamase drug-resistant gene. The method comprises the following steps: extracting bacterial plasmids; the method comprises the following steps of: downloading sequences of TEM-1 and ampC from GenBank, comparing nucleotide sequences by utilizing Megaalign, finding out a gene conserved region, and designing a primer by utilizing Primer expansion V5; and adding reactants of a PCR reaction system by taking the extracted whole-genome DNA of the bacteria and the extracted plasmid DNA of the bacteria as templates, and carrying out PCR amplification according to the reaction temperature required by the corresponding amplified substances. According to the invention, plasmid-mediated beta-lactam drug resistance detection and analysis are carried out on bovine-derived escherichia coli, so that drug resistance detection rates of TEM-1 type and ampC type genes of a whole genome are 76% and 95% (n is equal to 38); the drug resistance detection rates of the plasmid DNA are 98% and 89% (n = 44), and the drug resistance detection rates of the plasmid DNA are both 100%. The invention provides a scientific basis for further prevention and control of bovine-derived bacterial infectious diseases and reasonable use of antibacterial drugs, and also provides valuable data support and theoretical reference for guiding livestock farm medication and veterinary clinical scientific medication.

The invention discloses a reagent for nucleic acid extraction and amplification of porcine epidemic diarrhea virus and application, a nucleic acid rapid extraction technology part is a nucleic acid rapid extraction reagent, and the reagent mainly comprises tris (hydroxymethyl) aminomethane (Tris), TritonX-100 (TritonX-100) and chelating resin 100 (Chelex100). The PEDV on-site rapid detection technology comprises an RT-RAA reaction system and a colloidal gold test strip. The RT-RAA reaction system is composed of an RT-RAA nucleic acid amplification reagent, a pair of primers and a probe, and the primers and the probe are used for identifying an N gene conserved region of PEDV. The kit can complete virus nucleic acid extraction and nucleic acid in-vitro amplification and detection within 30 min, the specificity is good, and the detection sensitivity is 101 copies / microliter. The kit disclosed by the invention has the characteristics of simplicity in operation, rapidness and sensitivity, and is suitable for on-site extraction of a large batch of samples and on-site rapid diagnosis of PEDV (Porcine Epidemic Diarrhea Virus).

The invention provides a non-lineage animal pairing evaluation method based on genome data, and belongs to the technical field of bioinformatics and protection genetics, the method comprises the following steps: firstly, obtaining whole genome sequencing data of all individuals in a population, and carrying out quality control to obtain a high-quality SNP site set; aiming at all possible male and female pairing combinations in the population, calculating an inter-parent genetic coefficient depKin, an inter-parent heterozygous difference ratio HDR and an inter-parent potential risk load index GRLI, and predicting a fixed proportion Proh of ROH of offspring; performing z-score standardization processing on the four genetic indexes: performing principal component analysis on standardized data, extracting the first two principal components PC1 and PC2, determining a weight according to a variance contribution rate, and calculating a pairing comprehensive score; and generating a pairing candidate recommendation list for each individual according to the comprehensive score, and screening a high-quality pairing scheme. The method disclosed by the invention can be completely independent of pedigree records, and genetic evaluation is directly carried out based on genome data.

The invention belongs to the technical field of biological detection, and provides a bovine viral diarrhea virus, bovine enterovirus and bovine parvovirus triple PCR detection method which comprises the following steps: comparing nucleotide sequences of BVDV, BEV and BPV to obtain a gene conserved region, and constructing three pairs of specific primers; rNA of BVDV, BEV and BPV cell viruses is extracted respectively; taking the cDNA of the BVDV and the BEV and the DNA of the BPV as templates, and respectively carrying out single PCR (Polymerase Chain Reaction) amplification by utilizing the constructed three pairs of specific primers; carrying out plasmid construction and identification on the BVDV, the BEV and the BPV; establishing and optimizing a triple PCR reaction system; according to the invention, a triple PCR detection method is constructed through 5 'UTR gene sequences of BVDV and BEV and a VP2 gene sequence of BPV, and infection conditions of three cattle pathogens can be mastered only through one-time detection, so that the detection efficiency is greatly improved, the sample treatment time is shortened, and the detection cost is reduced.

The invention relates to sgRNAs for demethylation editing of an fgf2 gene of oryzias melastigma, an editing system and application, and belongs to the technical field of gene editing, and the sgRNAs are sgRNA1 + sgRNA2 + sgRNA3 or sgRNA2 + sgRNA3 + sgRNA4 + sgRNA5; the system comprises a Fuw-UbC-dCas9-tet1 plasmid and a pGL3-U6-sgRNAs plasmid, and the sequence of a conserved structural domain of a tet1 gene is as shown in SEQ ID NO. 1 (sequence identifier number 1). Through co-transfection of Mix1 or Mix2, the DNA methylation level of the fgf2 gene promoter can be effectively reduced by about 20.41% at most, the off-target efficiency is extremely low, the cell growth speed is remarkably increased, and stable inheritance is achieved.

The invention discloses a specific primer group, a kit and a method for detecting double photosynthetic bacteria, and relates to the technical field of microbiological detection. The specific primer group is designed based on a conserved region of a photosynthetic reaction center L subunit gene (pufL) of rhodopseudomonas and ectothiorhodospirillum, the conservative property is greater than or equal to 99%, the similarity rate is greater than or equal to 98.5%, specific amplification of target bacteria can be realized, and cross amplification interference is avoided. The matched kit comprises the primer group, 2 * SYBR Green qPCR Master Mix, a positive control component, a negative control component and the like, and the detection method realizes rapid and sensitive detection of the two photosynthetic bacteria by optimizing sample pretreatment, PCR amplification conditions and result interpretation standards. The detection specificity is high, the sensitivity is high (the lowest detection limit reaches 10 < 2 > cfu / mL level), the detection period is short (1-2 hours), the applicability is wide, the method can be used for detecting various samples such as river water, aquaculture water and microbial inoculum fermentation liquor, one set of primers can detect two target bacteria at the same time, the detection cost and the operation complexity are reduced, and the method is suitable for aquaculture environment monitoring and microbial inoculum application quality control.

The invention discloses a universal one-step RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection primer and a kit for goose astrovirus, and belongs to the field of biotechnology and diagnostic detection. The primer comprises an upstream primer GAstV-F and a downstream primer GAstV-R, the sequences of the upstream primer GAstV-F and the downstream primer GAstV-R are respectively shown as GAstV-F / R. The kit comprises the primer. According to the GAstV-1 and GAstV-2 gene conserved regions, universal detection primers are designed, each reaction condition of an RT-PCR system is optimized, a universal one-step RT-PCR detection method for detecting GAstV is established, and the universal one-step RT-PCR detection method has the characteristics of specificity, sensitivity, rapidness, accuracy and the like, and can be used for laboratory diagnosis, customs quarantine and molecular epidemiological research of GAstV infection.

The application belongs to the technical field of nucleic acid detection methods, and particularly relates to a rapid detection method of avian influenza virus, which comprises using a primer pair and a probe designed for a conserved region of M gene of AIV, and amplifying amplification reagents containing a sample to be detected based on super-fast qPCR technology, wherein the sequence of a forward primer in the primer pair is 5'-GGCCCCCTCAAAGCCGA-3'; the sequence of a reverse primer is 5'-CGTCTACGCTGCAGTCC-3'; and the sequence of the probe is ROX-CACTGGGCACGGTGAGCGT-BHQ2. The method of the application can rapidly and accurately detect whether the avian influenza virus exists in the sample to be detected and the content of the avian influenza virus, and has a good application prospect.

The invention relates to a primer probe combination for identifying E, J and K subgroups of avian leukosis viruses, a detection method and application of the primer probe combination. The detection system for identifying the avian leukosis virus subgroup comprises an upstream primer ALV-E-F, a downstream primer ALV-E-R and a probe ALV-E-P of a gp85 gene conserved region of targeted ALV-E, and an upstream primer ALV-J-F, a downstream primer ALV-J-R and a probe ALV-J-P of an env gene conserved region of targeted ALV-J, the upstream primer ALV-K-F, the downstream primer ALV-K-R and the probe ALV-K-P are used for targeting a gp85 gene conserved region of ALV-K. The detection system for identifying the avian leukosis virus subgroups has no cross reaction with other ALV subgroups and common avian pathogens, and has strong specificity.

The application relates to the technical field of animal virus molecular detection, and particularly discloses a multiple detection method for ASFV, PCV2 and PCV3 suitable for rapid real-time fluorescent PCR. Specific primers and TaqMan probes are designed according to the B646L gene of ASFV and the ORF1 gene conservative regions of PCV2 and PCV3, and the sequences are shown as SEQ ID NO:1-9. The primers and probes are optimized: avoiding strong GC anchoring at the 3' end of the primers, continuous AT series and G base at the 3' end of the probes, and shortening the length of the probes to adapt to rapid real-time fluorescent PCR. The minimum detection limit of the three viruses is not higher than 5 copies per muL, the detection is completed within 30 minutes, there is no cross reaction with 11 common pig pathogenic agents, the coefficient of variation is less than 2%, the sensitivity is high, the specificity is strong, the method is rapid and convenient, and is suitable for rapid diagnosis of mixed infection of ASFV, PCV2 and PCV3.

The invention discloses a dual digital PCR (polymerase chain reaction) detection method for monkey campylobacter jejuni and monkey enterocolitis yersinia, and relates to the technical field of microbiological detection, the detection method comprises the following specific steps: multi-source heterogeneous data collection: collecting monkey feces and intestinal tissue samples, extracting genome DNA (deoxyribonucleic acid) by using a phenol-chloroform method, detecting the concentration and purity of DNA, and detecting the concentration and purity of the DNA; after reaching the standard, storing at-20 DEG C for later use; according to the method, high-sensitivity and specific detection on the two pathogenic bacteria is realized through a combination of well-designed primers and probes, according to the method, a gyrA gene conserved region of monkey campylobacter jejuni CJ and a POL gene conserved region of monkey yersinia enterocolitica YE are used as detection targets, and specific fragments of 249bp and 182bp are amplified respectively; the method effectively avoids interference of non-specific amplification, can simultaneously detect two pathogenic bacteria in a single reaction system through a dual digital PCR technology, accurately calculates target gene copy number concentration through a Poisson distribution formula, and improves detection accuracy and reliability.

The invention belongs to the technical field of molecular biological detection, and particularly relates to an RT-LAMP (Reverse Transcription Loop-Mediated Isothermal Amplification) and immunochromatography combined PEDV (Porcine Epidemic Diarrhea Virus) nucleic acid detection kit and application thereof. According to the kit, the high sensitivity and high specificity of the loop-mediated isothermal amplification technology and the rapid and visual interpretation advantages of the immunochromatography test strip are combined, and on-site, rapid and accurate detection of the PEDV is achieved. The core of the invention comprises a set of specific primer group designed for a PEDV N gene conserved region, an optimized RT-LAMP reaction system and an immunochromatography test strip which is specially treated and is used for detecting an LAMP amplification product. The kit does not need an expensive thermal cycler, the whole detection process is completed within 40 min, the sensitivity can reach 100 copies / [mu] L, and the kit is particularly suitable for rapid diagnosis and epidemic prevention and control of sites such as pig farms and grassroots veterinary stations.

This invention belongs to the field of molecular biology detection technology, specifically relating to a Nipah virus (NiV) nucleic acid detection kit combining RT-LAMP and immunochromatography, and its application. This kit combines the high sensitivity and specificity of loop-mediated isothermal amplification (LAMP) technology with the rapid and visual interpretation advantages of immunochromatographic test strips, enabling on-site, rapid, and accurate detection of NiV. The core components of this invention include: a set of specific primers designed for conserved regions of the NiV gene, an optimized RT-LAMP reaction system, and specially treated immunochromatographic test strips for detecting LAMP amplification products. This kit eliminates the need for reverse transcription of RNA virus nucleic acids and expensive thermal cycling equipment. The entire detection process is completed within 20–30 minutes, with a sensitivity of up to 300 copies / μL. This solution combines the high efficiency of RT-LAMP technology with an intuitive, equipment-free detection method, achieving truly rapid and accurate on-site diagnosis of NiV.

The present invention relates to a set of nucleic acids, a kit and a detection method for detecting human adenovirus type 55 by the ERA-CRISPR / Cas12a method. The set of nucleic acids contains ERA probe HAdv55BP, forward primer F6, reverse primer R5 and CrRNA. The kit contains the set of nucleic acids. The detection method uses the kit. The present invention uses the conserved sequence of the Hexon gene of human adenovirus type 55 as a target, uses the above set of nucleic acids and uses the ERA-CRISPR / Cas12a method to effectively detect human adenovirus type 55, the results are reliable, the cost is relatively low, and it is particularly suitable for on-site or field instant detection.

The present application relates to the technical field of plant gene function, and aims to provide a construction method of a homologous silencing system of a Paeonia lactiflora bud dormancy gene, comprising the following steps: designing amplification primers according to gene annotation and a conserved sequence region in a Paeonia lactiflora bud dormancy full-length transcriptome, and constructing a conserved sequence fragment of a target gene into Agrobacterium tumefaciens containing a pTRV vector; taking a rhizome of a treated Paeonia lactiflora annual seedling as an infection material, and placing the infection material in Agrobacterium tumefaciens liquid containing pTRV1, pTRV2 and pTRV2-target gene fragments to perform infection treatment; potting the rhizome and moving the rhizome into a culture room to perform culture, observing phenotype and expression amount differences between the rhizome and a control group in a growth process, and confirming that the homologous silencing system is successfully constructed. The present application first completes miniaturization treatment and batch bacterial liquid infection of an underground rhizome system of Paeonia lactiflora, establishes a virus-mediated homologous function silencing system of a Paeonia lactiflora bud dormancy gene, and realizes the purpose of verifying functions of bud dormancy and even germination and growth related genes.

The invention discloses a monkey pox virus rapid detection method based on a microdroplet digital PCR technology. The monkey pox virus rapid detection method comprises the following steps: acquiring gene sequence data of monkey pox virus, analyzing a gene conserved region, and designing a specific primer and a probe based on the gene conserved region; establishing reaction conditions based on a fluorescent PCR detection system, and analyzing the sensitivity of the specific primer and the probe based on the reaction conditions to obtain an analysis result; selecting the same specific primer and probe based on the analysis result, and setting the same reaction condition to establish a digital PCR detection system; setting condition parameters, and verifying the sensitivity of the specific primer and the probe based on the condition parameters; adjusting condition parameters based on the verification result, generating an optimal reaction condition, detecting the detected sample based on the optimal reaction condition to obtain a detection result, and generating a monkey pox virus detection report; the monkey pox virus nucleic acid in an import and export sample is detected by using a digital PCR method, the sensitivity is higher, and the detection result is more accurate.

The invention discloses a sequence combination, a kit and a method for co-detecting a new bunyavirus and an anaplasma phagocytophilum based on RPA-CRISPR, and application of the sequence combination, the kit and the method. Comprising a first RPA primer pair and a first crRNA which are designed according to an SFTSV L segment conserved region, a second RPA primer pair and a second crRNA which are designed according to an AP 16S rRNA gene conserved region, an optimized universal T7 auxiliary primer, a kit prepared by using the sequence combination, and a detection method for detecting the new bunyavirus and the anaplasma phagocytophilum by using the kit. According to the detection method disclosed by the invention, an RPA amplification method and a CRISPR detection method are combined to detect the new bunyavirus and the anaplasma phagocytophilum at the same time, and the bottlenecks in co-amplification efficiency and orthogonal detection sensitivity in the prior art can be effectively overcome.

The invention discloses a fluorescent MIRA kit and method for detecting a monkey varicella virus, and relates to the technical field of biology.The kit comprises a primer probe combination, a buffer solution A, a buffer solution B, freeze-dried enzyme powder and a nucleic acid extraction.The primer probe combination is optimally designed and synthesized on the basis of a monkey varicella virus ORF21 gene conserved sequence; according to the present invention, the monkey varicella virus detection method is established and optimized, and the reaction system is further constructed, such that the optimal reaction condition and the optimal primer pair are determined, the monkey varicella virus detection method is established and optimized, and the constructed detection method has advantages of high sensitivity, good specificity, simple operation, fast detection speed, low cost, and wide application prospect. The kit has an important application value in detecting whether a sample contains the monkey varicella virus or not.

The invention discloses a coloration LAMP (loop-mediated isothermal amplification) rapid detection method for nocardiasis of snakehead. The method comprises the following steps: (1) extracting a to-be-detected snakehead tissue DNA (deoxyribonucleic acid) sample; (2) adding the DNA sample into an LAMP reaction system, wherein the reaction system comprises a specific primer group, Bst DNA polymerase, dNTPs, a reaction buffer solution and a color indicator hydroxycalcein, wherein the specific primer group is designed aiming at a nocardia argus 16S rRNA gene conserved region; (3) reacting for 20 minutes at the constant temperature of 65 DEG C in 25 microliters of sample adding system; and (4) after the reaction is finished, if nocardia argus exists in the sample, the reaction liquid becomes green, and if nocardia argus does not exist, the reaction liquid becomes orange. The method is simple and convenient to operate and suitable for convenient and rapid detection in production. The whole LAMP detection time is shortened to 30 minutes, and the detection can be completed; compared with a commercially available kit, the speed is increased by 41.59%; the primers are high in specificity, the sensitivity of the primers is 10-100 times higher than that of conventional PCR, low-concentration pathogenic bacteria can be effectively detected, and the primers are suitable for on-site rapid detection and large-scale screening in nocardiasis of channa argus and fry production.

The invention discloses a primer probe combination and application thereof in simultaneous identification of chlamydia psittaci and chlamydia abortus, and relates to the technical field of animal molecular biology test methods and test reagents. The method comprises the following steps: firstly, designing a universal primer probe for a common conserved region of chlamydia parrot (15 genotypes) and chlamydia abortus ompA genes, then designing an identification primer probe for conserved but different regions of ompA genes of two chlamydia, respectively marking FAM and VIC fluorophores by the two probes, then screening out an optimal primer probe combination and optimal PCR amplification reaction conditions, and finally identifying the ompA genes of the chlamydia parrot (15 genotypes) and the ompA genes of the chlamydia abortus. Finally, the sensitivity, specificity, repeatability and the like of the primer probe combination are verified through experiments. By using the detection method disclosed by the invention, the chlamydia psittaci and the chlamydia abortus can be identified at the same time, all genotypes of chlamydia psittaci can be detected, and the effect is obviously superior to that of the existing known detection method.

The invention discloses a detection system for visually detecting a swine Japanese encephalitis virus based on an RT-RAA-CRISPR / Cas12a system, and the visual detection of the JEV is realized by designing crRNA and RAA amplification primers for a JEV NS1 gene conserved region and triggering the breakage of a double-labeled ssDNA reporter molecule by using the specific recognition and trans-cleavage activity of Cas12a protein to an RAA amplification product. The lower detection limit of the method is 2.04 * 10 < 0 > copies / [mu] L; the clinical sample detection rate is 61.0% and is equivalent to that of 63.4% of PCR (Polymerase Chain Reaction). Therefore, the invention establishes a nucleic acid diagnosis method with strong specificity, high sensitivity, short detection time and simple operation, and the method is also suitable for pig farm on-site epidemic situation screening, animal quarantine ports, grassroots veterinary stations, remote area public health monitoring and other aspects.

The invention is applicable to the technical field of biological detection, and provides a primer probe group, system and method for simultaneously detecting mycoplasma pneumoniae and chlamydia pneumoniae, specific primers and probes are designed aiming at conserved regions of 16S rRNA gene of MP and MOMP gene of CP, the two probes are respectively marked with FAM and ROX fluorophores, and the specific primers and probes can be used for simultaneously detecting mycoplasma pneumoniae and chlamydia pneumoniae. Parallel detection of MP and CP nucleic acids can be realized in a single reaction tube. The detection system provided by the invention comprises the PCR enhancer, so that the detection sensitivity and the amplification efficiency are effectively improved. The method further comprises an optimized nucleic acid extraction step and a reaction procedure, has the advantages of being easy and convenient to operate, rapid in detection, high in sensitivity and high in specificity and is suitable for early-stage rapid diagnosis of clinical respiratory tract samples.

The invention belongs to the field of CRK gene family research, and discloses an identification and expression regulation analysis method of amorphophallus konjac AkCRKs gene family members, which is based on a hidden Markov model of a CRK gene conserved domain, and uses TBtools software to carry out identification and bioinformatics analysis on the AkCRKs gene family members. And the expression characteristics of AkCRKs gene family members in different tissue parts, jasmonic acid, soft rot bacteria (Pcc) and under low temperature stress are analyzed. 19 AkCRKs gene family members are identified in the whole genome of amorphophallus konjac and are non-uniformly distributed on 7 chromosomes. The promoter of the AkCRKs gene family member comprises a plurality of adversity stress response, hormone response, growth and development regulation and other cis-acting elements. The expression of the AkCRKs gene has tissue specificity and participates in response to Pcc, MeJA and low-temperature stress. The research result of the invention provides a basis for further researching the function and adversity stress response mechanism of the amorphophallus konjac AkCRKs gene.

The invention provides a national standard chicken gene residue detection method and application, and relates to the technical field of gene residue detection. The primer group comprises a primer pair and a DBQ double-quenching probe, wherein the primer group and the ND1 gene conserved region of the national standard chicken have specificity. The primer group adopted by the invention has remarkable technical advantages in detection of animal-derived components of livestock and poultry, and an efficient, specific and sensitive novel method is provided for food safety detection.

The invention discloses a crRNA group and a primer group for simultaneously detecting EV71 and CVA16 and application of the crRNA group and the primer group, and belongs to the technical field of gene detection. The crRNA group and the primer group are designed aiming at conserved regions of EV71 and CVA16 genes, and the nucleotide sequence of the EV71-crRNA is shown as SEQ ID NO.11; the nucleotide sequence of the CVA16-crRNA is as shown in SEQ ID NO. 12; the nucleotide sequences of the EV71 primer pair are as shown in SEQ ID NO. 1 and SEQ ID NO. 6; and the nucleotide sequences of the CVA16 primer pair are as shown in SEQ ID NO. 9 and SEQ ID NO. 8. The kit prepared by the invention comprises the crRNA group and the primer group, and further comprises a probe group, a Cas12a protein, a Cas13a protein and RNA extraction kit, an in-vitro constant-temperature RPA reverse transcription kit and a colloidal gold immune test strip. According to the invention, the kit is combined with an RPA / Crispr Cas12a / 13a system to realize simultaneous detection of EV71 and CVA16 by a one-tube method, and the sensitivity and the specificity are high; the detection result is presented by a fluorescence method, and is directly subjected to color development through test paper by utilizing a lateral flow chromatography technology, so that the operation is quicker and more convenient.

The invention discloses a method and a device for generating a siRNA precursor double-stranded RNA sequence aiming at a target organism in the field of bioinformatics and application of the siRNA precursor double-stranded RNA sequence. The invention aims to solve the technical problem of how to design a precursor double-stranded RNA (Ribonucleic Acid) aiming at specific siRNA (Small Interference Ribonucleic Acid) of a target species. The method comprises the following steps: analyzing a candidate conserved gene set of a target species based on transcriptome and protein data, and performing gene conservative analysis to obtain a target gene of the target species; then intercepting all precursor double-stranded RNA sequences based on transcriptome data of a target gene, and enumerating all available sequence fragments to obtain a candidate double-stranded RNA set 1; and screening from the candidate double-stranded RNA set 1 according to a preset off-target threshold to obtain a candidate double-stranded RNA set 2. The invention can be applied to RNAi prevention and control of pathogens or pests of agricultural related species.

The invention discloses a monkey pox virus triple fluorescence PCR nucleic acid detection kit, a detection method and application. The kit adopts a single-tube triple detection system and comprises (a) a monkey pox virus universal detection primer probe group and a targeted F3L and G2R gene conserved region; (b) a Clade Ib subtype detection primer probe group which targets D14L and A36R gene deletion feature regions; (c) a Clade II subtype detection primer probe group, which is used for targeting J2R and D18L gene feature sequences; and (d) an internal standard detection group which targets the human RNase P gene. By optimizing the concentrations of the primers and probes and four-channel fluorescent labels (FAM universal type, VI-Clade Ib, ROX-Clade II and CY5 internal label), single-tube synchronous detection is realized. The leak detection risk is reduced by adopting a double-target redundancy design, virus screening and subtype identification (Clade Ib / II) can be completed through single detection, and an internal standard system monitors the quality of a sample in the whole process. The kit is suitable for clinical early diagnosis, epidemic prevention and control and strain traceability.