39 results about "Rift Valley fever virus" patented technology

The present invention describes a reverse genetic system for Phlebovirus such as Rift Valley fever virus. This system comprised of RNA expression plasmids and protein expression plasmids. Additionally, the present invention also discloses the modification of this system to generate a recombinant virus that expresses a non-viral foreign gene. Furthermore, the present invention discloses the use of this system in the development of anti-Rift Valley fever virus vaccines, screening of antivirals testing for anti RVF immune response and developing marker vaccines for Rift Valley fever virus. We also claim the utility of this approach to other phleboviruses.

The invention provides a multiplex fluorescent polymerase chain reaction (PCR) kit and primers for detecting Ebola viruses, Marburg viruses, Lassa viruses and Rift Valley fever viruses. The multiplex fluorescent PCR kit comprises conventional reagents of an RT-PCR buffer and an RT-PCR enzyme mixed liquor and also comprises primers and probes for detecting the four viruses, wherein the primers are shown in sequences of SEG ID NO: 1-13 and the probes are shown in sequences of SEQ ID NO: 14-18. The multiplex fluorescent PCR kit, the primers and the probes realize rapid and accurate detection of pathogens of Ebola hemorrhagic fever, Marburg hemorrhagic fever, Lassa fever and Rift Valley fever, prevent the four infectious diseases from spreading into or out of the frontier port, are accurate and effective, have strong operability, and can be used for detection of the infectious diseases. Through the multiplex fluorescent PCR kit, the primers and the probes, suspect cases can be found timely and a capability of preventing the infectious diseases from spreading into our country is improved.

The present invention is directed to an artificial nucleic acid, particularly to an artificial RNA, and to polypeptides suitable for use in treatment or prophylaxis of an infection with a virus of the order Bunyavirales, particularly Severe fever with thrombocytopenia syndrome virus (SFTSV), Rift Valley fever virus (RVFV), or Crimean-Congo hemorrhagic fever virus (CCHFV), or a disorder related to such an infection. The present invention further concerns a Bunyavirales vaccine, particularly a SFTSV, RVFV, or CCHFV vaccine. The present invention is directed to an artificial nucleic acid, polypeptides, compositions and vaccines comprising the artificial nucleic acid or the polypeptides. The invention further concerns a method of treating or preventing a disorder or a disease, first and second medical uses of the artificial nucleic acid, polypeptides, compositions and vaccines. Further, the invention is directed to a kit, particularly to a kit of parts, comprising the artificial nucleic acid, polypeptides, compositions and vaccines.

The invention relates to a hemorrhagic fever associated pathogen identifying gene chip; the preparation method comprises preparation of a specific primer, preparation of a pathogen specific oligonucleotide probe, preparation of an oligonucleotide chip, establishment of an RT-PCR (reverse transcription-polymerase chain reaction) system and establishment of a hybrid system and a signal detection method. The gene chip prepared by the invention can be used for simultaneously identifying 16 hemorrhagic fever associated pathogen microorganisms, including Zaire Ebola virus, Sudan Ebola virus, marburg virus, lassa virus, junin virus, Machupo virus, rift valley fever virus, Crimea-Congo hemorrhagic fever virus, plasmodium, hantaan virus, SFTS (severe fever with thrombocytopenia syndrome) virus, dengue virus, yellow fever virus, Chikungunya virus, influenza A virus and influenza B virus. The gene chip has the characteristics of being rapid and accurate, high in throughput and high in sensitivity; and a new technological means is offered for the diagnosis of hemorrhagic fever pathogen, health supervision and the control and prevention of infectious diseases.

The invention discloses a fulminating-infectious-disease pathogen detecting primer pair and a kit. The primer pair comprises at least a pair of RT-LAMP primers of Ebola viruses, Lassa fever viruses, Marburg viruses, rift valley fever viruses, yellow fever viruses and Chikungunya fever viruses. By means of the primer system, the amplification reaction background is reduced, and sensitivity and specificity are quite good. The kit formed by the primer pair further comprises detecting liquid and a micro-fluidic chip; as an independent RT-PCR secondary amplification step of a detecting liquid system is omitted, detecting time is shortened; as the denaturation process and the renaturation process of nucleic acid do not exist, the polluted chance of RNA enzymes and the polluted chance of amplification nucleic acid are reduced, and the sensibility and the safety of detection are improved. By means of the constant-temperature sealed environment provided by a micro-fluidic chip system, rapid and constant-temperature amplification and automation result distinguishing of a nucleic acid extracting template are finally achieved, the requirement for test hardware is reduced, the use level of a reaction reagent is reduced, detection cost is reduced, and the result can be directly determined through color changes.

The invention discloses a new fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and a rift valley fever virus detection PCR system. The system consists of primers and a probe, Premix Ex Taq reaction solution, and sterile Tris water, wherein the primers and the probe have good specificity and high sensitivity, and are suitable for no cross reaction of the rift valley fever and Dengue virus.

The invention discloses a colloidal gold immunochromatographic test strip for detecting anti-RVF virus antibodies in human and animal serum, belonging to the technical field of biological detection. The test strip contains a colloidal gold-labeled specific RVF virus recombinant antigen glass fiber membrane, and a nitrocellulose membrane coated with a specific RVF virus recombinant antigen detection line and coated with a murine anti-RVF virus recombinant antigen monoclonal antibody quality control line. The colloidal gold immunochromatographic test strip disclosed by the invention not only has the advantages of easiness, convenience and quickness, high specificity, high sensitivity, accuracy and reliability, low cost and the like, but also can be used for reading results with naked eyes directly without special instruments, thus having very important medical value for the diagnosis, prevention and treatment of Rift Valley fever virus.

The invention provides primers and a probe for detecting the fragment S of a rift valley fever virus, wherein, a forward primer is 5'-GGATTACTTTCCTGTGATATCTGTTG-3'; a reverse primer is 5'-GTATCCTGGGAGGRCCATCWC-3', R refers to A or G and w refers to T or A); and a probe is 5'-F1-ACTCCACTGACACAACACGACGACCACT-Q1-3', F1 is a fluorescent reporter and Q1 is a fluorescent quencher. The invention also provides a real-time fluorescence RT-PCR (reverse transcription-polymerase chain reaction) detection method for the fragment S of the rift valley fever virus. In the method, pseudovirus grains of the rift valley fever virus are taken as a template, and the primers and the probe are used to carry out real-time fluorescent RT-PCR, thus judging whether the detected virus is the rift valley fever virus according to the detected result. The probe provided by the invention has high sensitivity and good specificity, and has no cross reaction with all of Africa swine fever virus, irides virus and goatpox virus.

The present invention describes subunit vaccines containing Gn and Gc glycoproteins of the Rift Valley Fever Virus, including nucleic acids encoding such glycoproteins, host cells, vectors, and immunoreagents generated with the glycoproteins, methods of vaccination, methods of diagnosis, and kits.

The invention relates to a rift valley fever virus RT-LAMP (reverse transcription-loop-mediated isothermal amplification) detection method which comprises the following detection process: step a, establishing an amplification reaction system which comprises 5 mul of a RNA sample, 2.5 mul of a Bst (bacillus stearothermophilus) DNA polymerase buffer (10x), 1 mul of a Bst DNA polymerase (8U/mul), 1 mul of AMV reverse transcriptase (10U/mul), 1 muld of NTP (25mmol/mul), 5 mul of Betaine (5 mumol/mul), 1 mul of MgSO4 (100nmol/mul), 1 mul of F3 and B3 (5pmol/mul ), 1 mul of BIP and FIP (70pmol/mul), and the balance of H2O for supplementing to 25 mul; wherein the F3,B3,BIP and FIP are four primers corresponding to L gene; step b, evenly mixing the components, reacting for 1.5-2H in water bath at 65 DEG C; and step c, using 1.5% agarose gel for electrophoresis, and then observing the results. The new rift valley fever virus RT-LAMP technology has the advantages of being fast, sensitive, simple in operation, high in safety, low in price, in no need of special equipment, and the like, particularly applicable to popularization and application in basic inspection and quarantine institutions, and provides a quick and practical diagnostic method for early diagnosis of rift valley fever virus.

The invention discloses rift valley fever virus humanized monoclonal antibodies and application thereof and belongs to the technical field of medicine. According to the antibodies, RVFV glycoproteinsGn and Gc expressed by baculovirus are adopted as antigens, memory B cells which can be combined with the RVFV glycoproteins Gn and Gc are screened from PBMCs of a patient suffering from rift valley fever, and through experiments such as single B cell sequencing, in-vitro neutralization and in-vivo protection, eight Gn humanized monoclonal antibodies capable of efficiently neutralizing RVFV infection and one Gc humanized monoclonal antibody are identified. The humanized monoclonal antibodies have very high in-vitro neutralization activity (IC50 can be as low as 1.93 +/- 0.6 pM and reaches a pMlevel), can effectively treat mice infected with RVFV, prevents the mice from being infected with RVFV, and has an extremely high application value on clinical treatment and prevention of the infection of RVFV.

The invention aims at providing a primer and probe for detecting rift valley fever virus by applying an RPA technology. The sequence of a forward primer body is showed in SEQ ID NO:1, the sequence of a reverse primer body is showed in SEQ ID NO:2, and the sequence of a probe is showed in SEQ ID NO:3. According to the primer and probe for detecting the rift valley fever virus by applying the RPA technology, a great number of RPA primers and probes are designed according to a rift valley fever virus genomic sequence, and a primer and probe combination which can rapidly and effectively detect rift valley fever virus nucleic acid can be screened out from the great number of the RPA primers and probes. Rapid detection is conducted by utilizing the set of the primer and probe, an obvious amplification curve can be obtained by taking rift valley fever virus nucleic acid RNA in vitro transcription as a positive control, and other pathogenic nucleic acids such as foot and mouth disease virus, contagious pustular dermatitis virus, mycoplasma capricolum and bluetongue virus which serve as a template to conduct the reaction do not have the amplification curve.

The invention discloses a triple detection method of arbovirus liquid chips. The method comprises the following steps: respectively designing specific primers and probes aiming at three types of arboviruses comprising Chikungunya viruses, Crimean-Congo hemorrhagic fever and Rift Valley fever viruses, determining the specificity of the primers and the probes via a single PCR and clone sequencing, and then carrying out multiplex-PCR amplification under the condition that the concentration ratio of the forward and reverse primers in an amplification reaction system is 1:1-1:8; then coupling the probes with fluorescent coding microspheres so as to prepare a microsphere mixing liquid and carrying out coupling quality control by coupling the probes; finally, carrying out a hybridization reaction on multiplex-PCR products and the microsphere mixing liquid, simultaneously adding streptomycin-phycoerythrin diluted by a nucleic acid detection buffer solution, and analyzing reaction products by utilizing a liquid chip detecting instrument. The method is good in detection specificity and can be used for simultaneously detecting the three types of arboviruses, and has high detection sensitivity due to the adoption of an asymmetric PCR amplification method in the process of the multiplex-PCR amplification.

The invention discloses a composition for preparation of a low-toxicity rift valley fever virus, a preparation method of the composition and an RVFV attenuated vaccine, and relates to the field of biotechnology. According to a reverse genetic system, introduction of excess basic groups is avoided, the primitiveness of a viral genome is kept to the greatest degree, and the reverse genetic system can stably express a low-toxicity rift valley fever virus strain in the later period, the defect of toxicity recovery is avoided, and the limitation is broken that by means of a traditional method, an attenuated strain can be obtained through continuous passage many times.

The invention provides a hybridoma cell strain capable of secreting an anti-rift valley fever virus NSs protein monoclonal antibody and an application thereof, and belongs to the technical field of biology. The preservation number of the hybridoma cell strain 2D6D10 capable of secreting an anti-rift valley fever virus NSs protein monoclonal antibody is CCTCC NO: C2019269. The invention further provides an anti-rift valley fever virus NSs protein monoclonal antibody secreted by the hybridoma cell strain and an application of the anti-rift valley fever virus NSs protein monoclonal antibody in preparation of a rift valley fever virus NSs protein antibody detection kit. The hybridoma cell strain 2D6D10 disclosed by the invention can generate a monoclonal antibody aiming at RVFV non-structuralprotein NSs protein, and the monoclonal antibody has a blocking effect, so that the blocking ELISA kit disclosed by the invention can be used for differential diagnosis of virus infection and inactivated vaccines or deletion vaccine immune antibodies.

The invention discloses a monoclonal antibody against Rift Valley fever virus. The antibody is obtained by screening with flow sorting-single cell PCR technology and has a unique CDR partition. The invention also discloses that the antibody is used in the preparation of Rift Valley fever. Application of heat therapy drugs. The monoclonal antibody disclosed in the invention has high-efficiency and specific anti-RVF activity, and also has the characteristics of high expression, high degree of humanization and good stability, and is suitable for industrial production.