WO2015053455A1 - Severe fever with thrombocytopenia syndrome virus, and sfts diagnostic method and kit using same - Google Patents

Abstract

Disclosed are: a severe fever with thrombocytopenia syndrome (SFTS) virus having - strand RNA sequence corresponding to + strand RNA sequence represented by each of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3; and a method and a kit for detecting SFTS virus through the detection of an antigen-antibody complex, using the SFTS virus. The SFTS virus in accordance with the present invention is a virus having a different genotype from those of previously identified viruses and can be usefully used in a diagnostic method, vaccine development, etc. for SFTS, which is a new epidemic against which, up to the present day, a vaccine has not yet been developed anywhere in the world.

Description

Severe febrile thrombocytopenia syndrome virus and SFTS diagnostic method and kit using same

The present application relates to the diagnosis of diseases using viruses.

Severe fever with thrombocytopenia syndrome (SFTS) is a serious disease with symptoms such as high fever, vomiting, diarrhea, thrombocytopenia, leukopenia, and multiple organ failure (6-30%). Yu XJ et al., N. Engl. J. Med. 2011; 364: 1523-32; Ding F et al Clin Infect Dis 2013; 56: 1682-3). SFTS is caused by the SFTS virus (HB29 strain) belonging to the field virus (Bunyavirus) and was first reported in China in 2011 (Yu XJ et al. Ibid ).

SFTS virus is mediated by Haemaphysalis longicornis , which is widely known in Korea (Chae JS et al. J Vet Sci 2008; 9: 285-93; Kim CM et al. Appl) Environ Microbiol 2006; 72: 5766-76).

Seroconversion and viremia of the SFTS virus have been found in breeding animals such as goats, sheep, cattle, pigs, and dogs, and these animals are thought to act as intermediate mediators in areas where the SFTS virus has spread (Zhao L et al. Emerg Infect). Dis 2013; 18: 963-5; Niu G et al. Emerg Infect Dis 2013; 19: 756-63).

Chinese Laid-Open Patent Publication No. 102618669 relates to the entire sequence of SFTS virus and its use, and discloses the entire sequence of SFTS virus identified in China.

Chinese Laid-Open Patent Publication No. 102070704 discloses a SFTS virus amplification and detection kit using SFTS virus.

There is a need for identification of new SFTS viruses and development of virus detection methods and vaccines using the same.

The present application is to provide methods, kits and vaccine compositions for diagnosing or detecting severe febrile thrombocytopenia virus.

In one embodiment the present application is directed to a severe condition in which L (Large), M (Medium) and S (Small) fragments have negative stranded RNA sequences that are inversely complementary to the DNA sequences represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively. Febrile thrombocytopenia syndrome (SFTS) virus is provided.

In another embodiment, the present disclosure also provides DNA molecules derived from SFTS of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3 from the Severe Thrombocytopenic Syndrome (SFTS) virus and each thymidine (T) in the sequence possessed by the DNA molecule. Provided is an RNA molecule derived from SFTS substituted with this uracil (Uracil, U), which corresponds to a plus stranded RNA or cRNA.

In another aspect the present disclosure also provides a polypeptide encoded by said RNA or DNA. In one embodiment according to the present invention the sequence of the polypeptide is SEQ ID NO: 4 (polypeptide encoded by L segment), SEQ ID NO: 5 (polypeptide encoded by M segment) and SEQ ID NO: 6 (L segment Polypeptide).

In another aspect, the present disclosure also provides a vector comprising the DNA sequence.

In another aspect the present disclosure provides prokaryotic or eukaryotic cells comprising the vector.

RNA or DNA derived from the SFTS virus according to the present invention can be usefully used for the production of primers, probes, etc. for the detection of SFTS virus or vector development for the production of RNA virus.

In another aspect, the present invention also provides a method of forming an antigen-antibody complex comprising contacting a sample of a virus comprising a viral antigen according to the present invention with a sample; And detecting the antigen-antibody complex, and provides a method for detecting severe febrile thrombocytopenia syndrome virus antibody or SFTS infection in vitro. The sample used in the method according to the present invention may be used one or more of whole blood, plasma or serum.

The method according to the invention can be used for quantitative or qualitative detection or diagnosis. In the method according to the present invention, the antigen-antibody complex detection may be detected through radioimmunoassay, Western blot, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence assay, and in one embodiment, the ELISA is a double antigen sandwich method. The antigen can be carried out using nucleocapsid protein of severe thrombocytopenia syndrome having the amino acid sequence of SEQ ID NO: 6 as an antigen.

In the method according to the invention the antigen may be labeled with a label selected from the group consisting of radioactive material, enzyme and fluorescent material.

In another embodiment, the present invention also provides a kit for detecting or diagnosing a severe febrile thrombocytopenia syndrome virus antibody or a diagnostic kit for a severe febrile thrombocytopenia syndrome virus infection, the kit comprising a virus sample comprising a viral antigen according to the present application and a reagent for detecting the antigen-antibody complex. It provides a kit comprising a. In the kit according to the present invention, the reagent for detecting an antigen-antibody complex includes a reagent for radioimmunoassay, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence assay.

In another aspect the present disclosure also provides immunogenic compositions for severe thrombocytopenic syndrome (SFTS) virus, comprising all or part of the virus according to the present application. Such compositions may further comprise an adjuvant.

The SFTS virus according to the present application is a virus having a genotype different from the SFTS virus previously identified in China. SFTS virus was first discovered worldwide in China in 2011, and has not been developed yet. It can be used for diagnosis or vaccine development.

Figure 1 is a result of observing the cytopathic effect observed in Vero cells by optical microscope, 200 times.

2 is an optical microscope for cytopathic effect observed in DH82 cells, The result was observed 400 times.

Figure 3 is a transmission electron micrograph of Vero cells (arrows) infected with SFTS virus according to the present application, the scale bar is 500nm.

Figure 4 is a phylogenetic analysis of the RNA-dependent RNA polymerase (RdRP) gene of SFTS virus according to the present invention compared to the virus isolated and identified in China and China. The analysis was performed using the neighbor-joining method, and the location of separation, year of separation, and GenBank access number were recorded. The length of the branch represents the evolution distance and the scale bar represents the 2.0% sequence distance.

FIG. 5 is a photograph of a microscope at 400-fold magnification of an immunofluorescence assay for detecting a virus antibody present in a serum sample of a patient using a virus cultured according to an example of the present application.

Configurations shown in the embodiments and drawings described herein are only one of the most preferred embodiments of the present invention and do not represent all of the technical spirit of the present invention, various equivalents that may be substituted for them at the time of the present application It should be understood that there may be variations and variations.

The present application is based on the identification of new severe febrile thrombocytopenia syndrome virus. Here we isolated and analyzed the virus that causes thrombocytopenia with fever, which is a virus of a genotype different from the virus isolated in China (Yu XJ et al., N. Engl. J. Med. 2011).

Thus, in one embodiment, the present application provides that L (Large), M (Medium) and S (Small) fragments have minus strand RNA sequences that are inversely complementary to the DNA sequences represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, respectively. Severe febrile thrombocytopenia syndrome (SFTS) virus.

The SFTS virus herein is a negative single stranded RNA virus belonging to the genus Bunyaviridae and phlebovirus. A spherical virus with a diameter of 80 to 100 nm, which is a small tick with a small tick. The genome is large (L). Medium (M). It consists of small (S) segments and contains RNA dependent RNA polymerase (RdRp), glycoprotein precursor (M), glycoprotein N (Gn), glycoprotein C (Gc), nucleocapsid protein (NP) and non-structural protein (NSs). Encode six proteins.

Negative or antisense strands (antisense to the sense or plus strands that encode a viral protein) are proteins or genes encoded as antisense, and after the sense or plus strand RNA generation for expression of the gene into the protein, translation is performed therefrom. Protein is produced.

The genome of the SFTS virus as identified herein is therefore RNA having a reverse complementary sequence to DNA corresponding to plus or sense strand RNA, represented by SEQ ID NOS: 1-3. For example, it can be obtained by reading the complementary sequence of the cDNA sequence represented by SEQ ID NO: 1 to 3 in the 5'-3 'direction, and replacing thymidine with uracil.

Viruses identified and separated herein have a genotype different from those previously identified, and may be useful for detecting SFTS virus and diagnosing SFTS virus infection using the same.

In this aspect, in another aspect, the present disclosure provides a method of forming an antigen-antibody complex by contacting a viral sample comprising a viral antigen according to the present application with a biological sample or a sample; And an in vitro severe thrombocytopenic syndrome virus antibody detection method comprising the step of detecting the antigen-antibody complex.

Viral antigens herein refer to all or part of a virus capable of generating an immune response against the viral antigen, wherein the inactivated whole virus, split virus, modified virus, virus-derived protein, viral derived glycoprotein, Viral derived surface proteins include, but are not limited to.

As used herein, a biological sample or sample refers to a substance or mixture of substances that includes one or more components capable of detecting a virus, and refers to cells, tissues or fluids derived from an organism, in particular human, such as whole blood, urine, plasma, and serum. Including but not limited to. It also includes cells or tissues derived directly from an organism as well as cultured in vitro. Various samples may be used for detection of the virus according to the present disclosure, but are not limited thereto. In one embodiment, whole blood, serum and / or plasma may be used. In other embodiments, tissue / cell or in vitro cell cultures obtained from organisms that have or are suspected of developing SFTS or are susceptible to SFTS virus infection or susceptible to infection may be used, but are not limited thereto. It also includes fractions or derivatives of the blood, cells or tissues. When using a cell or tissue, the cell itself or a fusion of the cell or tissue may be used.

Detection herein includes quantitative and / or qualitative analysis, including the detection of presence, absence, and detection of virus titer. Such methods are known in the art, and those skilled in the art will appreciate You will be able to choose the appropriate method.

Antigen-antibody complex detection in the methods according to the invention can be carried out through a variety of known methods, including, for example, radioimmunoassay, Western blot, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence.

In one embodiment, immunoassays such as direct ELISA, indirect ELISA, dual antigen sandwich ELISA, Enzyme Linked Immuno Sorbent Assay (RIISA) including competitive ELISA, and Radio Immuno Assay (RIA) are used. This method involves a biological sample on a first antibody bound to a solid substrate such as beads, membranes, slides or microtiterplates made of glass, plastic (eg polystyrene), polysaccharides, nylon or nitrocellulose. After the addition of a label, a label capable of direct or indirect detection may be labeled with a radioactive substance such as ³ H or ¹²⁵ I, a fluorescent substance, a chemiluminescent substance, hapten, biotin, digoxygenin, or the like, or the action of a substrate. Proteins can be detected qualitatively or quantitatively through binding of conjugated antibodies with enzymes such as horseradish peroxidase, alkaline phosphatase, and malate dehydrogenase, which are capable of developing or emitting light.

In one embodiment according to the invention a double antigen sandwich ELISA is used. In the double antigen sandwich method, an antibody is immobilized on a solid substrate, and then an antibody is bound thereto, and the antibody is bound to an antigen labeled with a fluorescent substance or biotin, and the antibody is detected. Proteins encoded by SFTS (including recombinant proteins) can be used, for example, nucleocapsid proteins having the amino acid sequence of SEQ ID NO: 6 can be used, in particular the SFTS virus according to the present invention can be used with the SFTS virus of the HB29 strain. It can be usefully used for differential detection.

Other immune response-based methods can be used, and in other embodiments, Ouchterlony plates, Western blots, Crossed IEs, Rocket IEs, Fused Rocket IEs, Affinity IEs, which can easily detect antibodies and / or antigens through antigen antibody binding. Immuno Electrophoresis can be used. Reagents or materials used in these methods are known and are for example antigen-antibody reactions, substrates that specifically bind antigens, nucleic acid or peptide aptamers, reactions with receptors or ligands or cofactors that interact with the complex. Can be detected or a mass spectrometer can be used. Reagents or materials that specifically interact with or bind to the antigen-antibody complexes of the present disclosure may be used in chip mode or with nanoparticles. The immunoassay or method of immunostaining is described in Enzyme Immunoassay, ET Maggio, ed., CRC Press, Boca Raton, Florida, 1980; Gaastra, W., Enzyme-linked immunosorbent assay (ELISA), in Methods in Molecular Biology, Vol. 1, Walker, JM ed., Humana Press, NJ, 1984 and the like. By analyzing the final signal intensity by the above-described immunoassay process, that is, by performing a signal contrast with a normal sample, it is possible to diagnose the infection.

In the method according to the invention the viral antigen may be labeled directly or indirectly in the form of a sandwich with a label selected from the group consisting of radioactive material, enzyme and fluorescent material. In the case of the direct labeling method, serum samples used in arrays and the like are labeled with a fluorescent label such as Cy3 or Cy5. In the case of a sandwich, an unlabeled serum sample is first detected by reacting with an array to which a detection reagent is attached, followed by binding to a target protein with a labeled detection antibody. In the case of the sandwich method, the sensitivity and specificity can be increased, and thus the detection can be performed up to pg / mL level. In addition, radioactive materials, coloring materials, magnetic particles and high-density electron particles may be used as the labeling material.

Fluorescence luminosity can be used with scanning confocal microscopy, for example Affymetrix, Inc. Or Agilent Technologies, Inc.

In another aspect the invention also relates to an in vitro severe thrombocytopenic syndrome virus antibody detection kit, the kit comprising a virus sample comprising the viral antigen according to the invention and a reagent for detecting said antigen-antibody complex.

The reagent for detecting an antigen-antibody complex included in the kit according to the present invention is a reagent used for radioimmunoassay, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence assay, and the above-mentioned reference is made.

In addition, as described above, the detection reagent may be directly or indirectly labeled in a sandwich form for detection of an immune response. In the case of the direct labeling method, serum samples used in arrays and the like are labeled with a fluorescent label such as Cy3 or Cy5. In the case of a sandwich, an unlabeled serum sample is first detected by reacting with an array to which a detection reagent is attached, followed by binding to a target protein with a labeled detection antibody. In the case of the sandwich method, the sensitivity and specificity can be increased, and thus the detection can be performed up to pg / mL level. In addition, radioactive materials, coloring materials, magnetic particles and high-density electron particles may be used as the labeling material. Fluorescence luminosity can be used with scanning confocal microscopy, for example Affymetrix, Inc. Or Agilent Technologies, Inc.

Kits herein may further include one or more additional ingredients required for binding assays, and may further include, for example, binding buffers, reagents for sample preparation, blood sampling syringes or negative and / or positive controls. .

The kit of the present disclosure, which may include the various detection reagents, may be provided for ELISA analysis, dip stick rapid kit analysis, microarray, gene amplification, or immunoassay according to an assay. The detection reagent may be selected according to the analysis mode.

In one embodiment, an ELISA or dipstick rapid kit is used, in which a viral antigen according to the invention can be provided attached to a substrate, for example a surface of a well or glass slide or a nitrocellulose in a multiwell plate. In the case of a dipstick, a technique widely used in the field of point of care testing (POCT), in which the viral antigen according to the present invention is bound to a substrate such as nitrocellulose, and when it is contacted with a sample such as serum, for example, a dipstick When one end of is contacted with a serum sample, the sample moves by virtue of capillary action to detect the substrate by binding to a viral antigen.

Viruses according to the invention can be developed into vaccines through appropriate treatment. In this aspect the present application relates to an immunogenic composition for severe thrombocytopenic syndrome (SFTS) virus, which comprises all or part of a virus according to the present application.

Viruses according to the invention for the preparation of immunogenic compositions are included attenuated or inactivated, methods for which are known (see Stanley A et al., Vaccine, 5 ^th Edition, Elsevier Inc. 2008).

The virus according to the present application may be administered prophylactically to adults or children who are primarily concerned with SFTS virus infection.

Immunogenic compositions according to the invention can be prepared using known methods. Those skilled in the art will be able to employ a variety of materials known for the preparation of the compositions in the preparation of immunogenic compositions according to the present disclosure. See, eg, Remington's Pharmaceutical Sciences (18th edition), ed. A. Gennaro, 1990, Mack Publishing Co., Easton, Pa. For example, the virus according to the present disclosure can be formulated using MEME (Minimum Essential Medium Eagle's Salt) containing 7.5% lactose and 2.5% human serum albumin or MEME comprising 10% sorbitol. The virus can also be used diluted with physiologically acceptable, sterile physiological saline.

Immunogenic compositions according to the invention can be administered using a variety of methods known in the art, and dosages can also be readily determined by one skilled in the art. For example, the virus according to the present invention may be a 0.1 to 1.0 ml sterile liquid solution containing 10 ² to 10 ⁷ infectious units (or plaque-forming units or tissue culture infectious doses). It may be formulated and administered by intramuscular, subcutaneous or intradermal route.

In addition, the composition according to the present application may be administered in a booster once a month or several months after initial prime administration, and will be readily determined by those skilled in the art.

Compositions according to the present disclosure may also further comprise an adjuvant. Those skilled in the art will be able to select the appropriate adjuvant known in the art. Adjuvant is used to improve the immunogenicity of the virus according to the present application, including but not limited to liposomes, synthetic adjuvants such as QS21, muramyl dipeptide, monophosphoryl lipid A, or polyphosphazine no. In addition, if necessary, genes encoding cytokines with adjuvant activity, such as GM-CSF, IL-2, IL-5, IL-12 or IL-13, can be inserted into the viral genome. It may also be introduced with other genes for enhancing immunogenicity, or with other genes for inducing cellular or humoral immunity.

Hereinafter, examples are provided to help understand the present invention. However, the following examples are provided only to more easily understand the present invention, and the present invention is not limited to the following examples.

Example

Example 1 Isolation of Viruses

The condition of patients who isolate the virus is as follows. The virus was isolated from a 63-year-old female patient living in Chuncheon, Gangwon-do, South Korea. The patient was reported to have been bitten by insects two weeks before the fever, and a month prior to the outbreak, no foreign travel or contact with livestock was confirmed. Diarrhea was followed six times a day from the 3rd day of onset, and on the 4th day, platelet and leukocyte reduction was observed. On the 10th day of onset he died of a number of organ failures.

Blood was collected at an anticoagulant state on day 8 of the patient and stored at -70 ° C. After 7 months, the blood was infected with Vero cells cultured in a monolayer (African Green Monkey Kidney, Korea Cell Line Bank KCLB No. 10081), and then cultured in RPMI1640 medium containing 2% FBS at 37 ° C. and 5% CO _2. It was. After 13 days, the culture was recovered and subjected to genetic analysis. In addition, the recovered medium was inoculated into DH82 cells (canine malignant histiocytosis macrophage, ATCC CRL-10389), and then cultured for 5 days, followed by cytopathic effect observed in cells infected with virus by optical inverted microscope. Was observed. Cytopathic effect observed in Vero cells is shown as deformation or destruction of normal cells, the cells fall out of the culture flask (Fig. 1). Cytopathic effects observed in DH82 cells resulted in virus-derived cells becoming larger than normal cells, transformed into cells containing a number of vacuoles, and extending pseudopods (FIG. 2).

Example 2 Virus Genetic Analysis

For morphological analysis of the virus, Vero cells infected with SFTS virus were fixed according to the previously disclosed method (Popov VL et al. J Med Microbiol 1995; 43: 411-21). After fixation, sections were made to a thickness of 65 nm using ultramicrotome (RMC MT-XL), stained with saturated 4% uranyl acetate and 4% red citrate, and then transmission electron microscope (HITACHI-7100, Hitachi High-Technologies, Japan) at 75 kV. The results are described in FIG.

Genetic analysis was performed as follows. RNA was extracted from the collected blood and Vero cells infected with the blood using a QIAamp Viral RNA Mini Kit (QIAGEN, Hilden, Germany) according to the manufacturer's method. The RNA was then used to amplify partial L fragments by performing reverse transcript PCR (RT-PCR) as previously described (Liu Y et al. Vector Borne Zoonotic Dis. 2012; 12: 156-60). . RT-PCR confirmed SFTS virus positive, and PCR products were analyzed by direct sequencing. Genotype AH12 (GenBank Accession no. HQ116417), the genotype showing the most similar nucleotide sequence among the nucleotide sequences of the LTS fragment and the common nucleotide sequence of the SFTS virus previously published in GenBank for the entire sequencing of the SFTS virus. A primer was designed using the base sequence of (Yu XJ et al., N Engl J Med 364; 16: 1523-1532.). RT-PCR was performed on each of the L, M and S fragments, and the PCR products were analyzed by direct sequencing. The end portion of each fragment was polyadenylation of the 3 'end of the cRNA, amplified cDNA end using a rapid amplification method, and then the base sequence was determined by direct sequencing. The cDNA sequences of L, M and S analyzed are represented by the sequences of SEQ ID NOs: 1, 2 and 3, respectively, which are described in GenBank in Accession Nos. Deposited as KF358691, KF358692 and KF358693.

Blast analysis (http://blast.ncbi.nlm.nih.gov/Blast.cgi) showed no analogous sequences other than SFTS virus. Homology Search Results The virus identified herein indicates that the previously identified SFTS virus and L, M, and S fragments have homology of 5.8% -99.8%, 94.1% -99.9%, and 94.8% -99.7%, respectively. appear.

Based on RNA-dependent RNA polymerase sequences, phylogenetic analysis of Chinese and Japanese SFTS viruses, known as neighbor-joining methods, is not identical to these viruses and has 95.9% -99.9% sequence relevance. It was shown (see Figure 2).

Example 3 Immunofluorescence Analysis Using Cultured Virus as Antigen

Another patient diagnosed with severe febrile thrombocytopenia syndrome was diagnosed by immunofluorescence, using the cultured virus as an antigen.

Specifically, patient serum was mixed 1: 3 in 3% bovine serum albumin to prepare a serum 1: 4 dilution solution, and serially diluted to prepare a 1:32 dilution solution. Diluted serum was dispensed in the order of 1: 4, 1: 8, 1:16, 1:32 in a slide coated with acetone-fixed virus, and finally, bovine serum albumin was dispensed as a negative control. It was. Immunofluorescence slides were reacted for 30 minutes in a 37 ℃ thermostat and then washed three times for 5 minutes with phosphate buffered saline. After drying the slide at room temperature, fluorescein isothiocyanate-conjugated chlorine antiphosphorus gamma immunoglobulin antibody IgM and IgG were dispensed, respectively, and reacted for 30 minutes in a 37 ° C thermostat, followed by three times of 5 minutes each with phosphate buffered saline. After washing, the solution was reacted with a fluorescent solution and then read under a microscope. The results are shown in FIG. 5, and as shown in FIG. 5, when the diagnosis was performed using immunofluorescence in the serum of convalescent patients using the cultured virus as an antigen, the antibody was strongly positive in the 1: 160 dilution serum for the IgG antibody. Confirmed.

Although the exemplary embodiments of the present application have been described in detail above, the scope of the present application is not limited thereto, and various modifications and improvements of those skilled in the art using the basic concepts of the present invention defined in the following claims are also provided. It belongs to.

All technical terms used in the present invention, unless defined otherwise, are used in the meaning as commonly understood by those skilled in the art in the related field of the present invention. The contents of all publications described herein by reference are incorporated into the present invention.

Claims (14)

1. Severe febrile thrombocytopenia syndrome (SFTS), in which L (Large), M (Medium), and S (Small) fragments have RNA sequences that are complementary to the DNA sequences represented by SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively. virus.
2. Contacting the virus sample comprising the viral antigen according to claim 1 with a sample to form an antigen-antibody complex; And detecting the antigen-antibody complex. Severe febrile thrombocytopenia syndrome virus antibody detection method in vitro.
3. The method of claim 2, wherein the sample is one or more of whole blood, plasma, or serum.
4. The method of claim 2, wherein the detection is quantitative or qualitative detection.
5. The method of claim 2, wherein the antigen-antibody complex detection is detected via radioimmunoassay, Western blot, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence.
6. The method of claim 5, wherein the ELISA is in a double antigen sandwich manner, wherein the antigen is a nucleocapsid protein of severe febrile thrombocytopenia syndrome having the amino acid sequence of SEQ ID NO: 6. 7.
7. The method of claim 2, wherein the antigen is labeled with a label selected from the group consisting of radioactive material, enzyme and fluorescent material.
8. Severe febrile thrombocytopenia syndrome virus antibody detection or diagnosis or Severe febrile thrombocytopenia syndrome virus infection diagnostic kit, the kit comprises a virus sample comprising a viral antigen according to claim 1 and the reagent for detecting the antigen-antibody complex, Kit.
9. The kit of claim 8, wherein the reagent for detecting an antigen-antibody complex is a reagent for radioimmunoassay, Enzyme linked immunosorbent assay (ELISA) or immunofluorescence assay.
10. An immunogenic composition for severe thrombocytopenic syndrome (SFTS) virus, comprising all or part of the virus according to claim 1.
11. The immunogenic composition of claim 10, wherein the composition further comprises an adjuvant.
12. An RNA molecule derived from SFTS, wherein the thymidine is substituted with uracil in any one of SEQ ID NO: 1 to SEQ ID NO: 3 derived from SFTS.
13. DNA molecule derived from SFTS represented by any one of SEQ ID NO: 1 to SEQ ID NO: 3.
14. A polypeptide encoded by the RNA or DNA molecule of claim 12.

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