168 results about "Viroid" patented technology

The present invention belongs to the field of biomedicine, and relates to polypeptides capable of inhibiting human coronavirus infections, particularly to polypeptides capable of inhibiting human coronavirus infections in a broad spectrum manner, and applications thereof. According to the present invention, the polypeptides capable of providing broad spectrum inhibition effects for infections caused by more than 2 human coronaviruses are provided based on the conservation property of the S2 region of a coronavirus S protein and the similar fusion mechanism; the test results show that the polypeptides can achieve the commonality of the human coronavirus, ie., the similar HR region and the same fusion mechanism mediated by the coronavirus S protein so as to provide a series of HCoV-EK polypeptides, wherein the polypeptides provide good inhibition effects for currently popular human coronaviruses, and further provide good inhibition activity for SARS virus (RsSHC014-CoV or RsW1V1-CoV) possibly infecting human; and the polypeptides of the present invention can provide the prevention and treatment candidate drug for the currently popular human coronaviruses and the novel human coronavirus possibly emerging in the future.

This invention relates to a gene-chip for plant virus detection, particularly it relates to a method, by utilizing gene-chip, to detct samples if it contains plant virus. It is used for novel nucleic acid primer of PCR, or, for detecting PCR product probe. This method can be used for detecting simultaneously determinand sample with 2-122 kinds of virus, sich as: AIMV, CMV, PVA, PVX, PVY, TMY, BBTV, LSV, SqMV, CEVD, ASGV, PXP, PVS, LMoV, DsMV, ZYMV, WMV, CRSV, CCCVd, PNRSV, SBMV, TRSV and viroid PSTVd etc.

The invention provides a three-color fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) combined detection method of enterovirus 71, Coxsackie virus A16 and other subtypes of enterovirus as well as a kit thereof. The method can rapidly and accurately detect the enterovirus 71, the Coxsackie virus A16 and the other subtypes of nucleic acids of enterovirus in a sample. The method comprises the following steps of: (1) acquiring and conveying a sample of an infected patient or a suspected patient; (2) preprocessing the sample and extracting RNA; (3) detecting the sample by adopting a one-step PCR-three-color fluorescent probe in-vitro amplification method; and (4) analyzing the corresponding sample according to the fluorescence intensity of each amplification reaction after the amplification reaction is finished, thereby judging the existence of the enterovirus 71, the Coxsackie virus A16 and the other subtypes of nucleic acids of the enterovirus in the acquired sample and being capable of carrying out accurate quantitation (a figure 3) on the enterovirus 71, the Coxsackie virus A16 and the other subtypes of nucleic acids of the enterovirus. The invention realizes the aim of carrying out rapid and accurate combined detection of the enterovirus 71, the Coxsackie virus A16 and the other subtypes of nucleic acids of the enterovirus.

The invention relates to a construction and an application of recombinant Chicken Marek's Disease Virus SC9-1 strain and SC9-2 strain. The recombinant virus construction method solves the technical problem that kanr gene cannot be knocked out again by a present method after kanr gene containing flp recognition sites at two ends is continuously used twice to knock out two specific functional genes on the same virus genome. The obtained recombinant virus MDV SC 9-1 strain and MDV SC 9-2 strain are used as production strains of Marek's Disease live vaccine. The prepared vaccine prevents the very virulent or emerging very virulent plus MDV induced chicken Marek's disease. The protective immunity effect of the vaccine is superior to CVI988/Rispens strain vaccine which is mostly widely used in foreign and domestic markets at present. The antigenicity of the recombinant virus is more similar to that of Chinese epidemic strain than the antigenicity of the similar virus rMd5deltameq which has been published in the United State. The strains provided by the invention will not induce tumor and has no immune suppression effect. Therefore, the strains are more applicable to China.

The invention discloses a triple RPA primer, probe, identification method and identification kit for rapidly screening and detecting a novel coronavirus (SARS-CoV2) and other SARS-similar coronaviruses. A first set of primers and probes (marked as FAM) are designed in a region of an N gene sequence of the SARS-CoV2 virus; a second set of primers and probes (marked as HEX) are designed in an E genesequence in a conserved region of an SARS-similar virus; and a third set of primers and probes (marked as Cy5, being different from the other SARS-similar coronaviruses) are designed in a specific region of an S gene sequence of the SARS-CoV2 virus. Compared with qPCR nucleic acid detection methods recommended, by the WHO and the national CDC, to be used in novel coronavirus clinics, the triple RPA primer, the probe, the identification method and the identification kit have the advantages that the sensitivity is high, the detection speed is high, and the novel coronavirus and the other SARS-similar coronaviruses can be effectively distinguished; and the nucleic acid detection time limit of the virus can be shortened from 90 minutes to 20 minutes, and rapid screening demands on the novel coronavirus and the other SARS-similar coronaviruses can be satisfied.

The invention discloses a degradable liquid mulching film and a preparation method thereof. The degradable liquid mulching film comprises the following raw materials by weight: 0.5-1.5 parts of polyvinyl alcohol, 1.5-4.5 parts of carboxymethylcellulose, 0.3-0.9 part of chitosan, 0.2-0.6 part of gelatin, and 0.05-0.10 part of humic acid. The degradable liquid mulching film has the characteristics of simple preparation method, low cost and convenient use, strong water absorption and water retention capacity, easily degradable components and no residue. Film mulching and cemented soil grains areutilized to reduce the evaporation of soil water, reduce the salt content of a plow layer, improve the physical and chemical properties of soil, and increase the soil temperature. The degradable liquid mulching film also has bactericidal effect and the capability of inhibiting plant viruses and viroids, and protects plants from the infection of bacteria and viruses, at the same time can promote the proliferation of beneficial bacteria in soil and obviously improve microfloras.

The invention relates to a method for purifying an escherichia coli expressed viroid particle. The purification method comprises a precipitation step, a first chromatography step, a second chromatography step, a concentration step and a third chromatography step, wherein the precipitation step comprises an ammonium sulfate precipitation step; the first chromatography step comprises a molecular screen chromatography step; the second chromatography step comprises a hydroxyapatite chromatographic analysis; the concentration step comprises an ultrafiltration and concentration step; the third chromatography step comprises a heparin affinity chromatography step.

The invention discloses a multiple-PCR (polymerase chain reaction) detection and warning method for sweet potato chlorotic stunt virus (SPCSV) and sweet potato twin virus (Sweepoviruses) in sweet potato roots. For the two types of viruses of SPCSV and Sweepoviruses, specific primer pairs CP-F and CP-R and primer pairs of VF and VR are designed, the detection primers are good in specificity, and only a stripe of the size of specificity can be amplified in a targeted virus sample. The detection method for SPCSV and Sweepoviruses is established by the aid of multiple-PCR primers, primary PCR reaction can be passed, two kinds of sweet viruses of SPCSV and Sweepoviruses can be detected whether to exist in the sweet potato roots or not, and quality of potato seeds is further judged; relation of sweet potato root virus with sweet potato seedling, field period virus symptoms and yield loss is determined, basis is provided for judging whether the potato seeds are qualified or not by detecting whether the potato seeds carry the virus or not, and great importance is achieved in quality of detoxified sweet potatoes and disease epidemic warning.

The invention discloses a degenerate reverse transcription-polymerase chain reaction (RT-PCR) detection reagent and a kit for hantavirus group. The detection reagent comprises a pair of degenerate heterozygous oligonucleotide primers, and the sequences of the degenerate heterozygous oligonucleotide primers are respectively G1: GCAACAGCAACATGGTTTcartaytayac and G2: CTTCTTCATTCATATTTCCATGCarnccyttytc; the non-merger consensus sequence of the 5' ends in the primers plays a role in stabilizing the combination of a 3' merger core area and a template under the condition that the degeneracy of the primers is not increased, so that the specificity of the degenerate PCR reaction is improved, various viruses in hantavirus can be amplified and detected, the homologous unknown viruses of the extendedgenes can also be detected and the amplified target fragments can be sequenced by the detection reagent; and the kit are relatively high in sensitivity and good in hantavirus group specificity, can be used for detecting domestic popular Hantaan and Seoul viruses, and can also be used for detecting other oversea epidemic strains. The degenerate RT-PCR detection reagent and the kit for the hantavirus group are high in sensitivity, good in hantavirus specificity and universal.

Disclosed are halogen containing nucleotide and nucleoside therapeutic compositions and uses related thereto. In certain embodiments, the disclosure relates to the treatment or prophylaxis of viral infections. Such viral infections can include tongaviridae, bunyaviridae, arenaviridae, coronaviridae, flaviviridae, picornaviridae, Eastern, Western, and Venezuelan Equine Encephalitis (EEE, WEE and VEE, respectively), Chikungunya fever (CHIK), Ebola, Influenza, RSV, and Zika virus infections.

The present invention provides a method for purifying recombinant coxsackievirus A16 (CA16) virus-like particles, an application thereof in an vaccine, and the vaccine. The method comprises carrying out high density fermentation culture of recombinantly expressed engineering bacteria and methanol-induced expression of CA16 viroid granulin, carrying out centrifugation for collecting thalli, performing high-pressure homogenization and breaking, and purifying the supernatant by ultrafiltration, ion exchange chromatography, hydroxyapatite chromatography and molecular sieve chromatography and the like. The CA16 viroid particle vaccine provided by the invention has good immunogenicity, safety, immunological characteristics and biological activity. The process is simple, and the purification is performed by chromatographic methods. The purification method is more conducive to linear amplification compared with density gradient centrifugation, and can be used in large-scale preparation and purification, and a high-purity (more than 99%) virus-like particle (VLP) stock solution can be obtained and can be used to prepare a vaccine for preventing CA16 infection, and has good economic value and application prospects.

The invention relates to a method for constructing a heart cell system of blotchy rockcod, which comprises: taking a heart tissue of the blotchy rockcod as a material, adopting a trypsin digestion method to start primary culture, and culturing the heart tissue in a DMEM/F12 liquid medium which contains fetal calf serum, basic fibroblast growth factors, I-type insulin-like growth factors, chondroitin sulfate and culture supernatant of fin cells of the blotchy rockcod in the log phase and has a pH value of between 7.0 and 7.4; and adopting the trypsin digestion method for subculturing. The heart cell system of the blotchy rockcod constructed by the method is subcultured for 70 generations currently. The technology is scientific and reasonable, is hopeful to be applied to separation and breeding of fish viroids, preparation of virus vaccines and research in the aspects of interaction of viruses and host cells, and so on, can also be taken as a model for researching environmental pollutants in environmental toxicology, and performs pollution monitoring and safety evaluation on various environmental pollutants in the sea such as genic toxins, mutagen, carcinogens, environmental hormones and endocrine disruptors.

The invention discloses a radix sophorae flavescentis and vinegar mixture for preventing and treating crop diseases. The mixture comprises a main agent and selectively added auxiliary agents, wherein the main agent is prepared from the radix sophorae flavescentis and fermented vinegar which are immersed for 30-40 days according to the weight ratio of 1:10-15, the immersing temperature is 4-30 DEG C, the radix sophorae flavescentis and the fermented vinegar are evenly mixed to prepare the main agent of the radix sophorae flavescentis and vinegar mixture, or the main agent is prepared by a decocting method; the auxiliary agents are prepared by organosilicone adjuvants, cell membrane stabilizers, folium isatidis decoction and water; the auxiliary agents and the main agent are combined to prepare different radix sophorae flavescentis and vinegar compositions, and one or a combination of more than one of the different radix sophorae flavescentis and vinegar compositions are used to form the radix sophorae flavescentis and vinegar mixture. Chinese herbal medicines, auxiliary agents and adjuvants, which have the function of inhibiting the growth and the development of fungi, bacteria, viruses, mycoplasma, chlamydia, viroid and other pests, are selected to be prepared scientifically; the mixture prevents and treats bacterial diseases, fungal diseases and viral diseases of crops and pests of trialeurodes vaporariorum, aphid, thrips and the like; the mixed formula can be used for increasing the crop immunity, and the purposes of healthy cultivation, disease prevention and pest control, quality improvement and efficiency improvement for plants are achieved.

The invention relates to a method for using low-temperature treatment to remove chrysanthemum virus. Wherein, it comprises low-temperature treatment, disinfection, cutting off and cultivating stem sharp, viroid detecting, cultivating neuter chrysanthemum. The invention can effectively remove chrysanthemum virus, with simple process and better effect.

The invention discloses an anti-foot-and-mouth disease type O virus-like particle vaccine and a preparation method thereof, relating to the field of genetic engineering and immunology. The invention provides a safe and reliable anti-foot-and-mouth disease type O virus-like particle vaccine capable of effectively preventing the viruses of foot-and-mouth diseases, a preparation method of the particle vaccine, an amino acid sequence and a deoxyribonucleic acid (DNA) sequence of the anti-foot-and-mouth disease type O virus-like particle vaccine, a safe and stable prokaryotic expression vector 28a-CP-VP1 and the application of the anti-foot-and-mouth disease type O virus-like particle vaccine. The anti-foot-and-mouth disease type O virus-like particle vaccine has, shown in SEQ ID NO.1, the amino acid sequence which forms the anti-foot-and-mouth disease type O virus-like particle vaccine, and can be expressed by escherichia col and independently packed to be a virus-like particle structure in escherichia coli cells and exist as the virus-like particles. The vaccine can induce animal to generate anti-foot-and-mouth disease antibodies at the same time after immunization; the preparation is safe; and the vaccine has no pathogenic action probably caused by attenuated vaccine and inactivated vaccine.

The invention relates to a preparation method of human papilloma virus virions. According to the method, human papilloma virus structural protein L1 and L2 are co-expressed in a methanol yeast expression system having a same double defect. An exogenous protein expression cassette used in the expression system comprises the following elements that: a) a marker gene sequence; b) a promoter region from methanol yeast comprises a promoter sequence b1 and the optional promoter upstream sequence b2; c) an exogenous protein DNA sequence; and d) a terminator region from the methanol yeast comprises a terminator sequence d1 and the optional terminator downstream sequence d2; wherein, the b) is located in the upstream of the c), and the d) is located in the downstream of the c); the marker gene sequence is selected from the coded methanol yeast LEU, URA, LYS, HIS or a ADE DNA sequence, and is located between b1 and b2 or d1 and d2; and the exogenous protein is human papilloma virus structural protein L1 or L2.

The invention relates to the field of biomedicine and discloses a method for fast acquiring recombinant HPV (human papilloma virus) L1 virus-like particles (VLP). The method is the optimization of a hydroxyapatite chromatography (HAP) method. The method includes: cells expressing the recombinant HPV L1 are broken to obtain coarse sample liquid, preliminary purification is performed, reducing agent is added to allow target protein components to depolymerize sufficiently and uniformly exist in a pentamer state, HAP is performed under a certain condition, and the target protein L1 can self-assemble into VLP during elution. By the method greatly contributive to the increasing of the yield of the target protein, target protein purity can be increased effectively, the virus-like particles can be obtained during chromatography, the special depolymerization and reassembling process is omitted, and the separation and purification process of the target protein L1 is allowed to be simple and easy to control and suitable for industrial production.

Described herein are modified influenza virus NS gene segments and nucleic acid sequences encoding such modified influenza virus NS gene segments. In certain embodiments, a modified influenza virus NS gene segment described herein comprises an influenza virus NS 1 open reading frame (ORF) lacking a stop codon, a heterologous nucleotide sequence, a 2A autoproteolytic cleavage site or another cleavage site, an NEP ORF, wherein the gene segment has one or more mutations in either the splice acceptor site, splice donor site, or both the splice acceptor and splice donor sites that prevents splicing of mRNA. Also described herein are recombinant influenza viruses comprising a modified influenza virus NS gene segment and the use of such viruses. The recombinant influenza viruses may be use in the prevention and/or treatment of influenza virus disease or as a delivery vector.