JP2005185235A - Method for diagnosis of susceptibility to viral infection and kit therefor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for the diagnosis of the susceptibility of an individual body to viral infection, especially infection with a virus having a mucin-type glycoprotein represented by Filovirus, and provide a kit for the diagnosis. <P>SOLUTION: The susceptibility of the individual body to the infection with the virus having a mucin-type (O-bonded) sugar chain is diagnosed by detecting the polymorphism of the 35th amino acid of the human macrophage galactose-type C lectin (hMGL), typically the Arg/Cys polymorphism of the amino acid. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a method and kit for diagnosing susceptibility to viral infection in mammalian individuals, including humans.

Filovirus infections against filovirus primates show very severe bleeding signs and high mortality compared to those caused by other hemorrhagic fever viruses. In virus Filoviridae causing the infection (Filoviridae), Marburg (Marburg) and two genera are known Ebola (Ebola), further the latter four species, namely, Zaire (Zaire), Sudan (Sudan), Ivory Coast and Reston. Of the four species, Zaire is considered the most highly toxic, and it has been reported that about 90% of infected people died (in the 1976 Zaire (now Congo) and Sudan epidemics) 430 of 500 people died). Regarding the course of infection by the virus, the incubation period after infection is 4 to 16 days, and headache and back pain begin immediately after the onset of illness. High fever, ocular conjunctivitis, pharyngitis, chest pain with cough, etc. occur around the second day after the onset of symptoms, followed by symptoms of vomiting and diarrhea, and sometimes bloody stool to tar stool. The patient rapidly became dehydrated, and bleeding tendency such as bleeding from gingiva, nose, vagina, subcutaneous, etc. from the 5th to 7th days, vomiting, melena, etc. Appears in sex. About half of the patients die from strong toxic symptoms and disseminated intravascular coagulation (DIC) on the 4th to 10th day after onset of illness.

  Against this background, Ebola virus infection, that is, Ebola hemorrhagic fever, is regarded as an international infectious disease that requires strict isolation treatment. Reston species have been reported to be less pathogenic than other Ebola species in non-human primate experiments, and in fact, no symptomatic infection has been confirmed in humans.

  In molecular biology, filoviruses are enveloped non-segmented single-stranded (-) polar RNA viruses that contain at least seven structural proteins. All of these proteins are translated from monocistronic polyadenylated mRNA transcripts. The fourth gene from the 3 ′ end of the filovirus genome encodes an envelope glycoprotein (GP) and is considered to be involved in receptor binding and fusion to the host cell membrane of the virus (Non-patent literature). 1 and 2). From the gene, an envelope GP and a non-structural secretory glycoprotein are expressed (Non-patent Documents 3 and 4). These glycoproteins are highly glycosylated and have an N-linked sugar chain and an O-linked sugar chain (mucin-type sugar chain). Depending on the cell line used, the terminal silylation pattern is different (Non-Patent Document 5).

  Recent studies on the infection of the filovirus have shown that dendritic cell specific ICAM-3 adherent non-Itegrin (DC-SIGN) or liver / lymph node specific ICAM-3 adherent non-Itegrin (L-SIGN) It was clarified that it plays an important role in infection. That is, Non-Patent Documents 6 to 9 show that when DC- or L-SIGN is overexpressed on the cell surface, these molecules recognize highly glycosylated Ebola GP, and the pseudotype of the virus. And showed that it promotes infection with genuine virus. In particular, high mannose-type N-glucan bound to viral GP is considered to be important in the interaction with lectins such as DC-SIGN and L-SIGN (Non-patent Document 10).

MGL
It is known that related cells derived from macrophages and myeloids express various lectins that recognize cell surfaces or soluble glycoproteins such as galectin-3, sialoadhesin, and mannose receptor. It has been. These lectins on macrophages are involved in cell-cell adhesion, hematopoiesis, endocytosis, and cleaning of necrotic cells and molecules. Macrophage galactose type C lectin (MGL) (Non-Patent Document 11) is one of the above-mentioned lectins, and is a type 2 transmembrane protein expressed on macrophages and immature dendritic cells. The inventors of the present invention have previously cloned the lectin in mice (Non-patent Documents 12 and 13). Furthermore, the gene products (including splicing variants) are in vitro, and the end of the galactose terminal glycoprotein is in vitro. It has been reported that it is involved in cytosis (Non-patent Documents 14 and 15).

  Human MGL (hMGL) has also been identified, and this lectin also recognizes a sugar chain having an O-linked galactose / N-acetylgalactosamine (ie, a mucin-type sugar chain) particularly strongly. In humans, this protein is expressed on monocyte-derived immature dendritic cells and macrophages, and has been reported to function as an endocytosis receptor for galactosylated glycoprotein antigens (Non-patent Document 12). And 16).

  In addition, Non-Patent Document 16 reveals the existence of a SNP in which the 35th amino acid located in the intracellular region of hMGL and adjacent to the transmembrane domain is changed from arginine (Arg) to cysteine (Cys). That is, it has been confirmed that in the exon 3 in the hMGL gene encoding the protein, there exists a SNP in which the codon corresponding to the amino acid is changed from CGC (Arg) to TGC (Cys).

  The hMGL has so far been confirmed to be the only C-type lectin specific to galactose / N-acetylgalactosamine on macrophages and to exhibit a ligand specificity different from that of the DC-SIGN and L-SIGN. (Non-patent Document 12).

However, there was no direct knowledge regarding the interaction between MGL and viral glycoprotein. In particular, considering the involvement of the above-mentioned DC-SIGN and L-SIGN in the filovirus infection process and the different ligand specificities between them, other lectin molecules, especially MGL molecules may be used in the virus infection process. There was no suggestion of involvement.
Takada, A., C. Robison, H. Goto, A. Sanchez, KG Murti, MA Whitt, and Y. Kawaoka, "A system for functional analysis of Ebola virus glycoprotein.", Proc. Natl. Acad. Sci. USA , vol. 94, pp. 14764-14769 (1997) Wool-Lewis, RJ and P. Bates, "Characterization of Ebola virus entry by using pseudotyped viruses: identification of receptor-deficient cell lines.", J. Virol., Vol. 72, pp. 3155-3160 (1998) Sanchez, A., SG Trappier, BW Mahy, CJ Peters, and ST Nichol, "The virion glycoproteins of Ebola viruses are encoded in two reading frames and are expressed through transcriptional editing.", Proc. Natl. Acad. Sci. USA, vol. 93, pp. 3602-3607 (1996) Volchkov, VE, S. Becker, VA Volchkova, VA Ternovoj, AN Kotov, SV Netesov, and H.-D. Klenk, "GP mRNA of Ebola virus is edited by the Ebola virus polymerase and by T7 and vaccinia virus polymerases." , Virology, vol. 214, pp. 421-430 (1995) Feldmann, H. and MP Kiley, "Classification, structure, and replication of filoviruses.", Curr. Top. Microbiol. Immunol., Vol. 235, pp. 1-22 (1999) Alvarez, CP, F. Lasala, J. Carrillo, O. Muniz, AL Corbi, and R. Delgado, "C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans.", J. Virol., Vol. 76, pp. 6841-6844 (2002) Baribaud, F., S. Pohlmann, and RW Doms, "The role of DC-SIGN and DC-SIGNR in HIV and SIV attachment, infection, and transmission.", Virology, vol. 286, pp. 1-6 (2001 ) Baribaud, F., S. Pohlmann, G. Leslie, F. Mortari, and RW Doms, "Quantitative expression and virus transmission analysis of DC-SIGN on monocyte-derived dendritic cells.", J. Virol., Vol. 76, pp. 9135-9142 (2002) Simmons, G., JD Reeves, CC Grogan, LH Vandenberghe, F. Baribaud, JC Whitbeck, E. Burke, MJ Buchmeier, EJ Soilleux, LJ Riley, RW Doms, P. Bates, and S. Pohlmann, "DC-SIGN and DC-SIGNR bind ebola gycoproteins and enhance infection of macrophages and endothelial cells. ", Virology, vol. 305, pp. 115-123 (2003) Lin, G., G. Simmons, S. Pohlmann, F. Baribaud, H. Ni, GJ Leslie, BS Haggarty, P. Bates, D. Weissman, JA Hoxie, and RW Doms, "Differential N-linked glycosylation of Human Immunodeficiency Virus and Ebola Virus envelope glycoproteins modulates interactions with DC-SIGN and DC-SIGNR. ", J. Virol., Vol. 77, pp. 1337-1346 (2003) Oda S., Sato M., Toyoshima S. and Osawa T., "Purification and characterization of a lectin-like molecule specific for galactose / N-acetylgalactosamine from tumoricidal macrophages.", J. Biochem. (Tokyo), vol. 104 , pp. 600-605 (1988) Suzuki N, Yamamoto K, Osawa T, Irimura T., "Molecular cloning and expression of cDNA encoding human macrophage C-type lectin: its unique carbohydrate specificity for Tn antigen.", J. of Immunology, vol. 156: pp. 128 -135 (1996) Tsuiji M., Fujimori M., Ohashi Y., Higashi N., Onami TM, Hedrick, SM, Irimura T., "Molecular cloning and characterization of a novel mouse macrophage C-type lectin, mMGL2, which has a distinct carbohydrate specificity from mMGL1. ", J. Biol. Chem., vol. 277, pp. 28892-28901 (2002) Denda-Nagai, K., Kubota N., Tsuiji M., Kamata M. and Irimura T., "Macrophage C-type lectin on bone marrow-derived immature dendritic cells is involved in the initialization of glycosilated antigens.", Glycobiology, vol. 12, pp. 443-450 (2002) Higashi, N., Morikawa A., Fujioka K., Fujita Y., Sano Y., Miyata-Takeuchi M., Suzuki N. and Irimura T., "Human macrophage lectin specific for galactose / N-acetylgalactosamine is a marker for cells at an intermediate stage in their differentiation from monocytes into macrophages. ", Int. Immunol., vol. 14, pp. 545-554 (2002) Higashi, N., K. Fujioka, K. Denda-Nagai, S. Hashimoto, S. Nagai, T. Sato, Y. Fujita, A. Morikawa, M. Tsuiji, M. Miyata-Takeuchi, Y. Sano, N Suzuki, K. Yamamoto, K. Matsushima, and T. Irimura, "The macrophage C-type lectin specific for galactose / N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells.", J. Boil. Chem ., vol. 277, pp. 20686-20693. (2002)

  An object of the present invention is to provide a method and a kit for diagnosing an individual's susceptibility to viral infection, particularly infection by a virus having a mucin-type glycoprotein typified by filovirus.

  In addition to DC-SIGN and / or L-SIGN, the involvement of MGL in the viral infection process was revealed. Furthermore, it has been found that the infection efficiency is affected by single nucleotide polymorphisms (SNPs) in MGL.

  Therefore, the present invention is characterized by detecting a polymorphism relating to amino acid No. 35 of human macrophage galactose type C lectin (hMGL), typically a CGC / TGC (Arg / Cys) polymorphism of the amino acid. Methods / kits are provided. As described above, the detection of the polymorphism is extremely useful in diagnosing an individual's susceptibility to infection by a virus having a mucin-type (O-linked) sugar chain.

  For example, the kit detects a SNP within the codon corresponding to amino acid 35 of hMGL, that is, CGC (Arg) / TGC (Cys) polymorphism in exon 3 of human macrophage galactose type C lectin (hMGL) gene. To include at least one primer and / or probe.

  Preferably, the primer or probe consists of at least 20 consecutive base sequences in hMGL gene exon 3 containing a codon corresponding to amino acid 35 of hMGL, or a sequence complementary thereto.

  Alternatively, the kit may include a restriction enzyme for detecting RFLP based on the SNP.

  Preferably, BsrB1 is used as the restriction enzyme.

  In particular, when the virus having a mucin type (O-linked type) sugar chain is Ebola virus, the kit of the present invention can be preferably used.

  The present invention diagnoses an individual's susceptibility to infection with a certain virus, typically a virus having a mucin-type (O-linked) sugar chain on its envelope GP, by detecting MGL SNPs. It is characterized by that.

  That is, the present invention demonstrates the significant involvement of MGL in the process of establishing infection with a virus having a mucin-type sugar chain, and that the MGL SNP against the virus infection can significantly affect the infection efficiency of the virus. Based on the clarification.

  More specifically, in the present invention, the polymorphism is represented by a SNP that causes substitution of Arg (CGC) to Cys (TGC) at a specific codon within exon 3 of the hMGL gene, which encodes amino acid 35 of hMGL. Based on the finding that the sensitivity of cells expressing the gene to the virus increases, substitution of the amino acid or SNP in the gene is detected.

  Here, the mucin-type sugar chain (also referred to as O-linked sugar chain) generally refers to a sugar chain in which the N-acetylgalactosamine is O-glycosidically bonded to the hydroxyl group of serine or threonine. . It is widely known that proteins present in virus particles can be modified by such sugar chains. Therefore, the term “virus having a mucin-type (O-linked type) sugar chain” used in the present invention includes Virus particles represented by envelope GP can include viruses modified with the mucin-type sugar chain. Examples of the virus include, but are not limited to, viruses of the family Filoviridae, that is, Ebola virus and Marburg virus.

  The hMGL gene is located on chromosome 17 and is composed of 9 exons. Up to now, expression of seven different subtypes has been known as a result of alternative splicing (Alternative Splicing). ing. These alternative splicing are known for exon 6 and / or exon 8, and the splicing variants are hMGL (6A), hMGL (6B), hMGL (6C), hMGL (6A / 8A), respectively. hMGL (6C / 8A) and hMGL (8A) are described.

  In exon 3, a SNP exists in the codon encoding the 35th amino acid of the hMGL protein (in this specification, the N-terminal methionine is counted as 1) (Higashi, N., K. Fujioka, K. Denda-Nagai, S. Hashimoto, S. Nagai, T. Sato, Y. Fujita, A. Morikawa, M. Tsuiji, M. Miyata-Takeuchi, Y. Sano, N. Suzuki, K. Yamamoto, K. Matsushima, and T. Irimura, "The macrophage C-type lectin specific for galactose / N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells.", J. Boil. Chem., vol. 277, pp. 20686-20693 (See 2002). That is, the C / T SNP is known at the 2108th base of the hMGL gene, and the 35th amino acid is substituted by Arg / Cys by the polymorphism. As shown in the examples below, the Cys polymorphism exhibits a significantly increased Ebola virus infectivity compared to the Arg polymorphism. Note that alternative splicing for exon 3 is not known.

  In the detection of the SNP, any method known to those skilled in the art can be applied as the method, and examples thereof include a PCR method, a NASBA (trademark) method, and an RFLP method.

  For example, using a sample of cells, body fluids such as blood, or tissue obtained from an individual whose sensitivity is to be diagnosed, and using genomic DNA extracted from the sample or mRNA from genomic DNA as a template, PCR or RT-PCR can be performed using an appropriate protocol. To detect the presence of either polymorphism.

In an example of RT-PCR using mRNA from a sample as a template, first, a corresponding cDNA is prepared by reverse transcriptase, and PCR is performed on the cDNA. For example, total mRNA is prepared from a sample, and cDNA is prepared with Moloney murine leukemia virus reverse transcriptase in the presence of dNTP, random hexamer, and the like. In PCR from the cDNA, hMGL cDNA is amplified with an appropriate pair of primers so as to give an amplification product containing exon 3 of hMGL. For example;
Sense: 5'-aatcacaccctccagacctccc-3 '(SEQ ID NO: 1)
Antisense: 5'-tcccaccaaaggcagctcagtg-3 '(SEQ ID NO: 2)
PCR amplification of the hMGL gene with a set of primers.

  Next, the PCR amplification product may be subjected to agarose gel electrophoresis and the like, and the nucleotide sequence of the amplification product may be determined. Preferably, however, it corresponds to the 35th amino acid of hMGL in exon 3 of the hMGL gene. The polymorphism is determined by a probe for detecting a single nucleotide polymorphism between CGC (Arg) and TGC (Cys) within the codon to be detected. The probe may be labeled or the like, and such a label is well known to those skilled in the art.

  Alternatively, PCR may be performed directly on the cDNA to amplify only one polymorphism, in which case the PCR primer is a CGC within the codon corresponding to amino acid 35 of hMGL in exon 3 of the hMGL gene. About single nucleotide polymorphism between (Arg) and TGC (Cys), it designs so that only either one may be selectively amplified. Furthermore, the NASBA ™ method may be applied directly to mRNA from a sample. For details of NASBA and its modification, refer to, for example, JP-A No. 05-219999. The NASBA primer also selectively selects only one of the single nucleotide polymorphisms between CGC (Arg) and TGC (Cys) within the codon corresponding to amino acid 35 of hMGL in exon 3 of the hMGL gene. Design to amplify. A primer / probe whose 3 'end corresponds to a polymorphic base is a preferred embodiment.

  In addition, the primer / probe sequence must have a sufficient length to ensure specific detection of the SNP. Usually, the length of the probe and primer should be at least 10 bases long, preferably 20 bases long, more preferably 30 bases long or longer, but for example several hundred bases The length will usually not be needed.

  For example, the probe may be prepared based on nucleotides 300 to 320 of hMGL cDNA (shown as SEQ ID NO: 3) or a sequence complementary thereto. The 310th base of the cDNA is the SNP to be detected (the 310th to 312th bases as a codon).

  A particularly preferred primer may have the 3 'end of the 310th base in the cDNA. In that case, selective amplification is possible simply by the fact that the 3 'end does not simply complement the template. For example, in SEQ ID NO: 3, the 290th to 310th base can be a suitable primer. In addition, refer to the description of Japanese Patent Application Laid-Open No. 08-501212 regarding the LCR method.

In addition, as a more preferable SNP detection of the present invention, an RFLP method using a restriction enzyme BsrB1 can be exemplified. That is, BsrB1 recognizes the sequence CCGCTC and restricts the sequence at the center. On the other hand, in the hMGL cDNA (SEQ ID NO: 3) in which the 35th amino acid is Arg, Since the sequence is also the C CGC TC, this gene produces a BsrB1 restriction fragment at that location. On the other hand, hMGL whose amino acid at position 35 is Cys has a base sequence from 309 to 314 as C TGC TC. Therefore, BsrB1 does not restrict the sequence, and therefore a restriction fragment different from the Arg-type hMGL genome. Give.

  More specifically, for example, the hMGL genome is PCR amplified with an arbitrary primer pair for specific amplification, the obtained PCR amplification product is restricted with the above BsrB1, and the restriction fragment is subjected to agarose gel electrophoresis to obtain the pattern. The SNP can be detected by comparison.

  In the above, all methods necessary for the amplification / detection protocol such as gel electrophoresis, blotting, hybridization, autoradiography, and scanning are well known to those skilled in the art. Therefore, it will be easy for those skilled in the art to construct a kit containing at least one of the probes or primers of the present invention including reagents according to such various protocols. For example, a DNA polymerase used for PCR amplification can be included in the kit.

  Without further explanation, those skilled in the art given the above explanation can make full use of the present invention. The following examples are given for illustrative purposes only.

I. Method (1) Virus Pseudophylovirus ^: A virus strain (VSVΔG ^* ) lacking the G protein of vesicular stomatitis virus (VSV G protein) and further modified to express GFP (Green Fluorescent Protein) Zaire, Sudan, Ivory Coast, Reston strain and Marburg virus) envelope glycoprotein (GP) was introduced (Takada, A., C. Robison, H. Goto, A. Sanchez, KG Murti, MA Whitt, and Y. Kawaoka, "A system for functional analysis of Ebola virus glycoprotein.", Proc. Natl. Acad. Sci. USA, vol. 94, pp. 14764-14769 (1997)).

  Each virus was grown in Vero E6 cells and stored at −80 ° C. until use.

(2) Cells Monkey kidney Vero E6 cells and human chronic myeloid leukemia cells (K562) were cultured in DMEM medium and RPMI 1640 medium supplemented with 10% fetal calf serum, L-glutamine and antibiotics, respectively.

  All infectious substances, including Ebola virus, were handled in a Biosafety Level 4 facility at the Canadian Science Center of Human and Animal Health.

(3) Expression of human MGL As a human MGL gene, the coding sequence of hMGL6A (Suzuki N, Yamamoto K, Osawa T, Irimura T., “Molecular cloning and expression of cDNA encoding human macrophage C-type lectin: its unique carbohydrate specificity for Tn antigen. ", J. of Immunology, vol. 156: pp. 128-135 (1996) and Higashi, N., K. Fujioka, K. Denda-Nagai, S. Hashimoto, S. Nagai, T. Sato, Y. Fujita, A. Morikawa, M. Tsuiji, M. Miyata-Takeuchi, Y. Sano, N. Suzuki, K. Yamamoto, K. Matsushima, and T. Irimura, "The macrophage C-type lectin specific for galactose / N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells. ", J. Boil. Chem., Vol. 277, pp. 20686-20693. (2002)), pRc, an expression vector for mammalian cells / CMV (manufactured by Invitrogen) was inserted in the sense direction (CR4 or CR4T) and the antisense direction (CR7: control). In CR4, the 35th amino acid is Arg, and in CR4T, the 35th amino acid gives Cys.

K562 cells were transformed with the above plasmid by electroporation, and transformants were selected with Geneticin (G418 sulfate, manufactured by GIBCO). In addition, hMGL6A ⁺ cells can be obtained from Higashi, N., Morikawa A., Fujioka K., Fujita Y., Sano Y., Miyata-Takeuchi M., Suzuki N. and Irimura T., “Human macrophage lectin specific for galactose / N-acetylgalactosamine is a marker for cells at an intermediate stage in their differentiation from monocytes into macrophages. ", Int. Immunol., Vol. 14, pp. 545-554 (2002). ) Were enriched using immunomagnetic beads prepared using Transformants were then cloned by limiting dilution, followed by infection with CR4T-1 (amino acid 35: Cys) and CR4-10 (amino acid 35: Arg) as MGL expressing cell clones and CR7 clones as controls. Used for experiments.

(4) Virus infection experiment for transformant 10 ⁵ cells / clone of K562 transformant (including CR7 clone as a control) were transformed into VSV (pseudotype Ebola virus: Zaire, Sudan, Ivory Coast, Reston and Marburg). The infection rate was evaluated by measuring the number of GFP positive K562 cells by FACS and standardizing the result for CR7 clone as 100.

II. Results Pseudofilovirus infection experiment The results of the infection experiment are shown in FIG. All of the pseudofiloviruses were found to exhibit a very high infection rate in the CR4T-1 clone, which is a transformant expressing hMGL (amino acid 35: Cys), as compared with CR7 as a control. The 35th amino acid Arg polymorphism (CR4-10) showed a reduced infection rate.

  The above results indicated that filovirus infections including Ebola are significantly affected by SNPs with substitution of amino acid 35 on hMGL.

  A method for diagnosing an individual's susceptibility to infection by a virus having a mucin-type (O-linked) sugar chain, comprising a polymorphism of amino acid No. 35 of human macrophage galactose type C lectin (hMGL), typically A method is provided which comprises detecting an Arg / Cys polymorphism of an amino acid. In particular, in one embodiment of the present invention, there is provided a kit for diagnosing an individual's susceptibility to infection by a virus having a mucin-type (O-linked) sugar chain, the kit comprising a human macrophage galactose type C-type lectin ( hMGL) including at least one primer and / or probe or restriction enzyme for detecting a CGC (Arg) / TGC (Cys) polymorphism in the codon corresponding to amino acid 35 of hMGL in exon 3 of the gene The kit is characterized.

FIG. 1 shows Zairo, Sudan, Ivory Coast, Reaston, and Marburg, which are firovirus pseudotyped viruses based on vesicular stomatitis virus (VSV) (Zaire, Sudan, Ivory Coast, Reston, and Marburg, respectively). GP expressing) hMGL expression transformant of human chronic myeloid leukemia cells (K562) (amino acid 35 is Cys: CR4T-1), and hMGL expression transformant (amino acid 35 is Arg: CR4-10), And the result of having measured the infection rate at the time of infecting control (CR7) by FACS is shown. In the figure, the infected cells are measured by FACS, and the background infection rate is subtracted from each data. In addition, the number of infected cells of CR7 as a control is normalized as 100, and the relative infection rate is expressed. Experiments were performed in triplicate and standard deviations are indicated by bars.

Claims (5)

To detect a single nucleotide polymorphism between CGC (Arg) and TGC (Cys) in the codon corresponding to amino acid 35 of hMGL in exon 3 of human macrophage galactose type C-type lectin (hMGL) gene,
The kit for detecting a single nucleotide polymorphism described above, comprising 1) at least one primer and / or probe; or 2) at least one restriction enzyme.   The kit according to claim 1, wherein the kit is used for diagnosing an individual's susceptibility to infection with a virus having a mucin-type (O-linked) sugar chain.   The kit according to claim 1 or 2, wherein the primer or probe comprises at least 20 or more consecutive base sequences in hMGL gene exon 3 containing a codon corresponding to amino acid 35 of hMGL or a sequence complementary thereto.   The kit according to claim 1 or 2, wherein the restriction enzyme is BsrB1.   The kit according to any one of claims 2 to 4, wherein the virus having a mucin-type (O-linked type) sugar chain is an Ebola virus.

Images