KR101850375B1

KR101850375B1 - A composition and kit for detecting a laminitis, method for detecting a laminitis and method for screening a therapeutic agent for a laminitis

Info

Publication number: KR101850375B1
Application number: KR1020150161038A
Authority: KR
Inventors: 류덕영; 김용백
Original assignee: 서울대학교산학협력단
Priority date: 2015-11-17
Filing date: 2015-11-17
Publication date: 2018-04-19
Also published as: KR20170057664A

Abstract

A composition, a kit, and a method for diagnosing an individual's flea can be used to efficiently diagnose the fleas of an individual. Further, according to the method of screening a therapeutic agent for cilia, a candidate substance capable of treating cilia can be efficiently screened.

Description

TECHNICAL FIELD The present invention relates to a composition and a kit for diagnosing fleas, a method of diagnosing fleas, and a method of screening a flea infusion agent.

Compositions and kits for diagnosing myplary fleas, methods of diagnosing mycorrhizal fungi, and methods of screening therapeutic agents of mycobacterial fleas.

It is a connective tissue that supports the hoof in the third phalanx of the alveolar process. It is a structure in which the epithelial layer protruded from the inside of the hoof wall and the dermal layer are interlocked with each other with the tough basement membrane interposed therebetween. Fig. 1 is a view showing the structure of a crockery. In case of phlebophilia, chronic bruxism is caused by a problem in the basement membrane, which is relatively weak in the connective structure, and the continuity between bone and hoof is lost. When the third phalanges breaks through the sole of the foot, it causes serious pain and lass.

The cause of cilia is not clear, but it is known to be caused mainly by gastrointestinal problems caused by carbohydrate hyperphagia. Short-chain carbohydrates migrate into the cecum without digestion in the stomach of the mammal and are fermented by Gram-positive bacteria in normal flora to acidify the environment of the hindgut. Acidified intestinal environment causes overgrowth and death of Gram-positive bacteria and endotoxin from dead bacteria is absorbed through the permeable barrier to cause systemic toxicosis. Toxicosis activates matrix metalloproteinase (MMP), an enzyme present in the half-attached spot of the basement membrane, to cause segregation of the basement membrane and dermis, do.

Although accurate diagnosis is essential to determine the feasibility and prognosis, the diagnosis of cilium salts still depends only on the cardiovascular index and clinical symptoms. Accordingly, there is a demand for biomarkers capable of specifically diagnosing bacterial infections and a diagnostic method using the same.

An aspect of the present invention provides a composition for diagnosing dysplasia of an individual.

Another aspect provides a kit for diagnosing dysplasia of an individual.

Another aspect provides a method of diagnosing dermatophytes in an individual.

Another aspect provides a method for screening a therapeutic agent for a cilia.

One aspect is the use of a serpin A3-8-like gene, a C4b-binding protein alpha chain gene or a keratin type II cytoskeletal 1-like gene -like gene of the present invention, which comprises a preparation for measuring the level of expression of the gene of the present invention.

The serpin A3-8-analog protein may have the sequence of NCBI Accession No: XP_001498757, GI number: 338719856 or SEQ ID NO: 1. The C4b-binding protein alpha chain protein may be one having the sequence of NCBI Accession No: XP_001492582, GI number: 149708028 or SEQ ID NO: 2. The keratin type II cytoskeleton 1-analog protein may have the sequence of NCBI Accession No: XP_001504496, GI number: 194212030 or SEQ ID NO: 3.

The increase or decrease in the expression of the serpin A3-8-analogue gene, the C4b-binding protein alpha chain gene or the keratin type II cytoskeleton 1-analog gene has been found by the present inventors to be associated with the phyllite. Thus, the expression level of the serpin A3-8-analog gene, the C4b-binding protein alpha chain gene or the keratin type II cytoskeleton 1-analog gene can be used to diagnose Diclofenac.

As used herein, the term "serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog" refers to a naturally occurring wild-type serine A3-8-analog, C4b- Alpha chain or keratin type II cytoskeletal 1-analog and functional variants thereof. Also, in the present specification, the term "serpin A3-8-like gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene" refers to a naturally occurring wild-type serine A3-8- , C4b-binding protein alpha chain or keratin type II cytoskeletal 1-analog, and a gene encoding a functional variant thereof.

The expression level may include any expression level of a gene encoding a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton 1-analog. The expression level may be an expression level at the mRNA or protein level. Thus, the composition may comprise an agent for measuring the amount of mRNA of serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog, amount of protein, or combination thereof have. The agent may be a substance that specifically binds to a serine A3-8-analog, a C4b-binding protein alpha chain, or a transcript of a keratin type II cytoskeletal 1-analog. The agent may be a primer, a probe, or an antisense sequence thereof that specifically binds to the mRNA of a serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog or its complementary sequence; Or a combination thereof. The agent may be a primer, a probe, or an antisense sequence thereof that specifically binds to a gene encoding a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton 1-analog; Or a combination thereof. The genes encoding the serpin A3-8-analog, the C4b-binding protein alpha chain, and the keratin type II cytoskeleton 1-analog may each have a nucleotide sequence encoding the amino acid sequence of SEQ ID NOS: 1, 2, have. The primer may be one which provides a polymerization initiation point in a polymerization reaction with a polymerase. The primer may be one used in a nucleic acid amplification reaction. The term "amplification" refers to increasing the number of copies of the target sequence or its complementary sequence. The nucleic acid amplification reaction can be performed by any method known in the art. Amplification of nucleic acids involves either a method requiring multiple cycles during amplification or a method performed at a single temperature. Examples of cycling techniques include those that require thermal cycling. Methods that require thermocycling include polymerase chain reaction (PCR). PCR is known in the art. PCR typically involves denaturing double stranded DNA to single stranded DNA by thermal degradation, annealing the primer to the single stranded DNA; And synthesizing a complementary strand from the primer. An isothermal amplification method is a method in which a single temperature or a major aspect of the amplification process is performed at a single temperature. In contrast to PCR, in which the reaction product is heated to allow additional primers to bind in order to separate the double strands, the isothermal method relies on a strand displacing polymerase to separate the double strands and rescopy the template do. Isothermal methods can be distinguished by methods that rely on substitution of primers to initiate reiterative template copying and methods that rely on sequential reuse or novel synthesis of single primer molecules. Methods that rely on primer substitution include helicase dependent amplification (HDA), exonuclease dependent amplification, recombinase polymerase amplification (RPA) and loop mediated amplification and loop mediated amplification (LAMP). Methods that rely on sequential reuse or novel synthesis of single primer molecules include those selected from the group consisting of strand displacement amplification (SDA) and nucleic acid based amplification (NASBA and TMA). The primers may be included in one, or two or more sets according to the amplification method selected. The primer may be a PCR primer.

The mRNA expression level is measured by the presence or absence of the mRNA of the serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeletal 1- It may be a measure of the amount of mRNA as a process of confirming the degree of expression. This can be measured by directly isolating the mRNA or by using a primer or a probe for the mRNA. Analysis methods include RT-PCR, competitive RT-PCR, real-time RT-PCR, RNase protection assay (RPA), northern blotting ), Nucleic acid microarrays comprising DNA, or combinations thereof. RT-PCR is a method of analyzing RNA, in which cDNA obtained by reverse transcription of mRNA is amplified by PCR and analyzed. After the RT-PCR, RT-PCR was performed to confirm the band pattern and the band thickness by using a pair of primers specifically prepared for the gene in the amplification step of the RT-PCR, thereby confirming the expression level and mRNA expression level of the gene And comparing it with the normal control, the presence or absence of the lyophilis of the individual can be easily judged.

As used herein, the term "primer" refers to a nucleic acid sequence having a free 3 'hydroxyl group, capable of forming a base pair with a template complementary to a specific nucleotide sequence, &Lt; RTI ID = 0.0 > and / or < / RTI > Primers can initiate DNA synthesis in the presence of reagents for polymerization (i. E., DNA polymerase or reverse transcriptase) and four different nucleoside triphosphates at appropriate buffer solutions and temperatures. For example, a sense and antisense primer having 7 to 50 nucleotide sequences is used as a specific primer for the mRNA of serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog gene PCR amplification is performed to determine the presence or absence of the presence of the third order lyophilis in the individual by measuring the amount of the desired product. The PCR conditions, the lengths of the sense and antisense primers can be appropriately selected according to techniques known in the art. The primers may be used in an amount of 10 to 100, 15 to 100, 10 to 80, 10 to 50, 10 to 30, 10 to 20, 15 to 80, 15 to 50, 15 to 30, 15 to 20, 20 to 100, 20 to 80 , 20 to 50, or 20 to 30 nt.

As used herein, the term "probe" means a target nucleic acid, for example, a nucleic acid fragment such as RNA or DNA capable of specifically binding to mRNA, and the presence, amount and amount of specific mRNA And the like. The probe may be prepared in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, an RNA probe, or the like. For example, hybridization is carried out using a probe having a sequence complementary to the mRNA of serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog polynucleotide, The presence or absence of the phagocytosis of the individual or the degree of the onset thereof can be confirmed. Selection of suitable probes and hybridization conditions can be appropriately selected according to techniques known in the art. The probes may be in the form of a probe having a size of 10 to 100, 15 to 100, 10 to 80, 10 to 50, 10 to 30, 10 to 20, 15 to 80, 15 to 50, 15 to 30, 15 to 20, 20 to 100, 20 to 80 , 20 to 50, or 20 to 30 nt.

In addition, the primer or probe can be chemically synthesized using a phosphoramidite solid support synthesis method or other well-known methods. Such nucleic acid sequences may also be modified through a variety of methods known in the art. Examples of such modifications include, but are not limited to, methylation, capping, substitution with one or more of the natural nucleotide analogs, or modifications between nucleotides such as uncharged linkers (e.g., methylphosphonate, phosphotriester, phosphoramidate, carbamate Etc.) or charged linkages (e.g., phosphorothioate, phosphorodithioate, etc.). The primer or probe may also be modified using a label that can provide a detectable signal directly or indirectly. Examples of such labels may include radioactive isotopes, fluorescent molecules, or biotin.

The agent may be a serine A3-8-analog, a C4b-binding protein alpha chain, or a keratin type II cytoskeletal 1-analogue protein or a peptide or protein that specifically binds to an mRNA encoding the same. The peptide or protein may be an antibody, a ligand, a receptor, an agonist, an antagonist, or a fragment thereof; Or a combination thereof.

The expression level of the protein is expressed in a serine A3-8-analogue gene, a C4b-binding protein alpha-chain gene or a keratin type II cytoskeletal 1-analogue gene in a biological sample to diagnose the presence or absence of infectious agent in the individual. It may be a process of confirming the presence and the degree of expression of the protein. For example, by directly isolating a serine A3-8-analog gene, a C4b-binding protein alpha chain gene or a keratin type II cytoskeleton 1-analog protein gene, or a serine A3-8-analog, a C4b- The serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeletal 1-analog protein is identified using an antibody or a fragment thereof that specifically binds to the keratin type II cytoskeletal 1- . Analysis methods include Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis Immunoprecipitation assays, Complement Fixation Assays, FACS, mass spectrometry, magnetic bead-antibody immunoprecipitation, protein chips, or a combination thereof. . For example, the ELISA may be a direct ELISA using a labeled antibody that recognizes an antigen attached to a solid support, an indirect ELISA using a labeled antibody that recognizes the capture antibody in a complex of antibodies recognizing an antigen attached to a solid support, A direct sandwich ELISA using another labeled antibody that recognizes an antigen in a complex of an antibody and an antigen attached to a solid support or an antibody that recognizes an antigen in a complex of an antibody and an antigen attached to a solid support, Lt; RTI ID = 0.0 > ELISA < / RTI > using a labeled secondary antibody that recognizes the antibody. In addition, an antibody is attached to a solid support, a sample is reacted, and a labeled antibody that recognizes an antigen of an antigen-antibody complex is attached to the antibody to produce an enzyme, or a secondary antibody labeled with an antibody recognizing an antigen of the antigen- The antibody can be detected by a sandwich ELISA method in which an antibody is attached and developed enzymatically, and the degree of complex formation between the protein and the antibody can be confirmed to confirm the presence or absence of the infectious agent in the individual.

Also, Western blot analysis using, for example, one or more antibodies to the protein may be used. The western blot analysis separates the whole protein from the sample, electrophoreses it, separates the protein according to its size, transfers it to the nitrocellulose membrane and reacts with the antibody. Then, the amount of the protein can be confirmed by confirming the amount of the produced antigen-antibody complex by using the labeled antibody, and the presence or absence of the infectious agent in the individual can be confirmed.

The detection method can be carried out by a method of examining the amount of protein expression in a normal control group that is not affected by phylloplasma and no other manipulation and the amount of expression of the protein in a biological sample, and the mRNA or protein level is It can be expressed as absolute (eg, μg / ml) or relative (eg, the relative intensity of the signal).

The term "antibody ", as the term is known in the art, refers to a specific immunoglobulin directed against an antigenic site. The antibody may specifically bind to a serpin A3-8-analog, a C4b-binding protein alpha chain, or a keratin type II cytoskeletal 1-analog or fragment thereof. A fragment of a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton 1-analog may be, for example, an immunogenic fragment. Wherein said fragment is selected from the group consisting of serpin A3-8-analog, C4b-binding protein alpha chain or serine A3-8 having one or more epitopes that can be recognized by antibodies to keratin type II cytoskeletal 1- - analog, a C4b-binding protein alpha chain or a fragment of a keratin type II cytoskeletal 1-analogue protein. A serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1-analog gene is cloned into an expression vector to form a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton Analogs, C4b-binding protein alpha chains or keratin type II cytoskeletal 1-analog proteins, which are encoded by the 1-mer-1-analog gene, and the resulting serine A3-8-analog, C4b-binding protein Antibodies can be prepared from alpha chain or keratin type II cytoskeletal 1-analog proteins according to conventional methods in the art.

The form of the antibody includes a polyclonal antibody, a monoclonal antibody, or a recombinant antibody, and includes all immunoglobulin antibodies. The antibody refers to a complete form having two full-length light chains and two full-length heavy chains. The antibody also includes a special antibody such as a humanized antibody. Polyclonal antibodies can be prepared by injecting an immunogen-causing biomarker protein or fragment thereof into an external host according to conventional methods known to those skilled in the art. The external host may use mammals such as mice, rats, sheep, rabbits. The immunogen may be administered with an adjuvant to increase antigenicity when administered by intramuscular, intraperitoneal or subcutaneous injection methods. Then, blood can be periodically taken from the external host to collect the serum exhibiting improved activity and specificity for the antigen, or isolate and purify the antibody therefrom.

Monoclonal antibodies can be produced by immortalized cell line generation techniques by fusion known to those skilled in the art. The method for producing the monoclonal antibody will be briefly described. The protein is purified and immunized with Balb / C mouse in an appropriate amount of about 10 占 퐂, or polypeptide fragments of the protein are synthesized and bound to bovine serum albumin to immunize mice Producing lymphocytes are fused with human or mouse myeloma to generate immortalized hybridomas, and only hybridoma cells that produce the desired monoclonal antibody are selected using the ELISA method to proliferate The monoclonal antibody can then be isolated and purified from the culture. Monoclonal antibodies can also be obtained by obtaining antibodies to the commercially available serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeletal 1-analog.

These antibodies can be used to detect the presence or absence of an enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), sandwich assay, Western blotting on polyacryl gel, or immunoblotting , It can be confirmed whether or not the protein is expressed in a biological sample.

The term "fragment " means, for example, a polypeptide that does not have the structure of an intact antibody, peptide or protein, but has a specific antigen binding site or binding domain directed against the antigenic site. Said fragment comprising a functional fragment of an antibody molecule that is not a complete form of the antibody having two light and two heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and may be Fab, F (ab ') 2, F (ab') ₂ or Fv. The binding fragment may comprise at least seven amino acids, for example, at least 9 amino acids, and at least 12 amino acids.

The expression level may be an expression level in a sample isolated from the subject. The sample may be separated from the dough. The sample may be blood, plasma, serum, urine, feces, saliva, tears, cerebrospinal fluid, cells, tissue or a combination thereof separated from an individual. The entity may belong to Perissodactypa . The entity may be an Equus genus . Equus can include horses, donkeys, or zebras. The horse is referred to as equus ferus . The above equus ferus ) species is Equus Peruvian Scavalus ferus caballus , Equus Perus ferus ferus , or Equus Perus, ferus przewalskii ) subspecies. The subject may include horses, cattle, buffalo, rhinoceros, or camels.

Another aspect provides a kit for diagnosing acute platelet deficiency in an individual comprising an agent for measuring the expression level of a serine A3-8-analog gene, a C4b-binding protein alpha chain gene or a keratin type II cytoskeletal 1- .

For the formulations "," individual "and" for diagnosing dysplasia "for determining the level of expression of the serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog, As shown in Fig.

The kit may be a microarray for diagnosing dysplasia capable of measuring the mRNA expression level of, for example, serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeletal 1- Lt; / RTI > The microfluorescent diagnostic microarray can be readily prepared by those skilled in the art according to methods known in the art using the serpin A3-8-analog, the C4b-binding protein alpha chain or the keratin type II cytoskeleton 1-analog have. The microarray is characterized in that the cDNA of the sequence corresponding to mRNA of the serine A3-8-analog, the C4b-binding protein alpha chain or the gene encoding the keratin type II cytoskeletal 1-analogue protein or a fragment thereof is attached to the substrate as a probe There may be something.

When the kit is used for measuring the expression level of mRNA of, for example, serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog gene, it is necessary to perform RT- Element. &Lt; / RTI > In addition to the respective primers specific for the mRNA of the marker gene, the RT-PCR kit can also include enzymes such as test tubes or other appropriate containers, reaction buffers, deoxyribonucleotides (dNTPs), Taq polymerase and reverse transcriptase, DNase, RNase Inhibitor, DEPC-water (dEPC-water), or sterile water. It may also contain a primer pair specific for the gene used as a quantitative control.

The kit may also include an antibody specifically binding to a serpin A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1-analog protein, a substrate suitable for immunological detection of the antibody, a suitable buffer, A secondary antibody labeled with an enzyme or a fluorescent substance, or a chromogenic substrate. The substrate may be a nitrocellulose membrane, a 96-well plate synthesized from polyvinyl resin, a 96-well plate synthesized from polystyrene resin, or a glass slide glass. As the chromogenic enzyme, peroxidase or alkaline phosphatase may be used. The fluorescent material may be FITC, or RITC. The chromogenic substrate may be ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), OPD (o-phenylenediamine) or TMB (tetramethylbenzidine).

Another aspect relates to a method of contacting a sample isolated from an individual with a substance that specifically binds to a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1-analog protein or an mRNA encoding the same, Forming a complex; Determining the level of expression of the serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeletal 1-analog gene in the sample by measuring the level of the complex; The expression level of the measured serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene was measured using the serpin A3-8-analog gene, C4b-binding protein Alpha chain gene and a keratin type II cytoskeletal 1-analog gene; And the expression level of the serpin A3-8-analog gene, the C4b-binding protein alpha chain gene and the keratin type II cytoskeletal 1-analog gene of the individual are changed in comparison with the expression level of the control group, The method comprising the steps of:

The method comprises contacting a sample isolated from an individual with a substance that specifically binds to a serpin A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1-analog protein or an mRNA encoding the same, To form a complex.

The entity may belong to Perissodactypa . The entity may be an Equus genus . Equus can include horses, donkeys, or zebras. The horse is referred to as equus ferus . The above equus ferus species is derived from Equus ferus caballus , Equus Perus ferus ferus ), or Equus Perus ferus przewalskii ) subspecies. The subject may include horses, buffalo, rhinoceros, or camels. The expression level may be an expression level in mRNA or a serine A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeleton 1-analog protein step. Thus, the measurement may be carried out in an amount of mRNA, a serine A3-8-analog, a C4b-binding protein alpha chain, or a keratin type II cytoskeletal 1-analog protein or a combination thereof. A serine A3-8-analog, a C4b-binding protein alpha chain, or a keratin type II cytoskeletal 1-analog and a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton The measurement of the amount of the 1-analogue protein is as described above. The sample may be blood, plasma, serum, urine, feces, saliva, tears, cerebrospinal fluid, cells, tissue or a combination thereof separated from an individual. The tissue may be loose.

The method also includes measuring the level of the complex to determine the level of expression of the serpin A3-8-analog, C4b-binding protein alpha chain and keratin type II cytoskeletal 1-analog in the sample.

Measuring the level of the complex may be accomplished by measuring the level of the detectable protein attached to a substance that specifically binds to the serpin A3-8-analog, the C4b-binding protein alpha chain, and the keratin type II cytoskeletal 1- And detecting the signal coming from the cover sheet. Measuring the level of the complex may include re-separating the complex to determine levels of serine A3-8-analog, C4b-binding protein alpha chain and keratin type II cytoskeletal 1-analog protein or mRNA encoding the same, or Determining the level of a substance that specifically binds to a serine A3-8-analog, a C4b-binding protein alpha chain and a keratin type II cytoskeletal 1-analogue protein or an mRNA encoding the same, It may be to measure the level. The measurements were performed using RT-PCR, competitive RT-PCR, real-time RT-PCR, RNase protection assay (RPA), Northern blotting, A nucleic acid microarray including DNA, Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunization Immunoprecipitation assay, Complement Fixation Assay, FACS, mass spectrometry, magnetic bead-antibody immunoprecipitation, protein chip, or a combination thereof. .

The method comprises measuring the level of expression of the serine A3-8-analog gene, the C4b-binding protein alpha chain gene and the keratin type II cytoskeleton 1-analog gene measured as the serine A3-8-analog gene, C4b-binding protein alpha chain gene and a keratin type II cytoskeletal 1-analog gene. The normal control group may be an individual not infected with leprosy.

The method also includes determining whether the level of expression of the serine A3-8-analog gene or the C4b-binding protein alpha chain gene measured is greater than the serine A3-8-analog gene or C4b-binding protein alpha Chain gene of the present invention is higher than the expression level of the chain gene, it may be determined that the subject is afflicted with infectious salt. The determining step further comprises determining the level of expression of the serpin A3-8-analog gene or the C4b-binding protein alpha chain gene measured in the normal control group or the serine A3-8-analog gene or the C4b-binding protein alpha chain Gene or keratin type II cytoskeletal 1-analog gene, the expression level of the gene is determined to be unaffected by phyllite.

The method may also include determining if the expression level of the measured keratin type II cytoskeletal 1-analog gene is lower than the expression level of the keratin type II cytoskeletal 1-analog gene measured in a normal control, It may be determined that it is caught by leaflet. In addition, the determining step may be such that when the expression level of the measured keratin type II cytoskeletal 1-analog gene is higher than or equal to the expression level of the keratin type II cytoskeletal 1-analog gene measured in the normal control group, It can be determined that it is not infected with leaf infestation.

The method may also include administering to the individual an amount of the compound of the present invention when the expression level of the serine A3-8-analog gene, the C4b-binding protein alpha chain gene and the keratin type II cytoskeletal 1- Lt; RTI ID = 0.0 > chlorophyll < / RTI > The expression level of each of the above-mentioned expression levels was 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900% and 1000% %, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% and 100%.

Another aspect includes the steps of administering a candidate agent to a subject; Contacting a sample isolated from said subject with a substance that specifically binds a serpin A3-8-analog, a C4b-binding protein alpha chain and a keratin type II cytoskeletal 1-analog protein or an mRNA encoding the same, ; Determining the level of expression of the serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeletal 1-analog gene in the sample by measuring the level of the complex; The expression level of the measured serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene was measured using a serine A3-8-analog gene, C4b-binding A protein alpha chain gene and a keratin type II cytoskeletal 1-analog gene; And the expression level of the serpin A3-8-analog gene, the C4b-binding protein alpha chain gene and the keratin type II cytoskeletal 1-analog gene of the individual are changed in comparison with the expression level of the normal control, The method comprising the steps of: determining the efficacy of the treatment of dermatophyte therapy;

The method comprises administering a candidate agent to a subject. The candidate material may be any substance expected to be effective in treating platelets. The route of administration of the candidate substance can be appropriately selected depending on the selected substance. The route of administration may be by any means, such as, for example, intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous. The material can be administered systemically or locally, for example, to a dough.

Contacting a sample isolated from said subject with a substance that specifically binds to a serpin A3-8-analog, a C4b-binding protein alpha chain and a keratin type II cytoskeletal 1-analog protein or an mRNA encoding the same, Measuring the level of the complex to measure the expression level of the serine A3-8-analog gene, the C4b-binding protein alpha chain gene or the keratin type II cytoskeletal 1-analog gene in the sample " And "the expression levels of the measured serpin A3-8-analog, C4b-binding protein alpha chain or keratin type II cytoskeletal 1-analog were measured using the serine A3-8-analog, C4b-binding protein alpha Chain or keratin type II cytoskeletal 1-analogue "is as described above.

The method comprises the steps of: when the expression level of the serine A3-8-analog gene, the C4b-binding protein alpha chain gene and the keratin type II cytoskeletal 1-analog gene of the subject is changed compared to the expression level of a normal control, And determining the substance to be effective for treatment of polyphlogy. The expression level of each of the above-mentioned expression levels was 1%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70% , 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900% and 1000% %, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% and 100%.

It is to be understood that the terms or elements mentioned in the description of the screening method mentioned above in the description of the method of diagnosis are as mentioned in the description of the method for diagnosing the above.

According to one aspect, a composition for diagnosing an organopharynx of an individual can be efficiently used for diagnosing a dysphonia of an individual.

In accordance with another aspect, the kit for the diagnosis of the dermatophyte of an individual can be effectively used to diagnose dermatophytes of an individual.

A method for diagnosing dermatophytes of an individual according to another aspect can be used effectively to diagnose dermatophytes of an individual.

According to another aspect of the method for screening a therapeutic agent for cilia, a candidate substance capable of treating cilia can be efficiently screened.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a view showing the position and shape of a croft leaf in a horse hoof. FIG.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example One. Leprosy Identification of proteins whose expression level in plasma was changed by induction

One. Cilia Inducing and Measuring Protein Levels Expressed in Plasma

(One) Experimental animal

In a study of artificial inflections, one horse ( Equus caballus ) were used. In the study of natural acupuncture induction, one horse (normal control) and two horses with naturally occurring phyllotaxis were used. All horses were Warmblood and used approximately 600 kg of horse radish.

(2) Leprosy Induction and evaluation method

Fructooligosaccharide (Sun Oligo P2, Kyowon, Korea) was used to induce artificial myofibroblasts. For the first 3 days, an induction dose of 1 g / kg was administered followed by an additional dose of inducing dose of 10 g / kg twice a day in Nasogastric tubes. After induction dose, body temperature, hoof heat, heart rate, and other clinical symptoms were measured and evaluated for 2 days. Hematologic, serological, and histologic examinations were performed after horses' euthanasia to evaluate artificially induced mycorrhizal infections. The evaluation of naturally induced mycorrhizas was assessed and diagnosed by a professional veterinarian in accordance with the rash grading system proposed by the American Veterinary Association.

(3) Plasma sampling method

The plasma of horse was collected from the jugular vein before administration of the induction dose of fructooligosaccharide, placed in BD vacutainer EDTA-K2 (10.8 mg, 6 ml), and centrifuged at 3000 rpm for 10 minutes using a centrifuge. After that, the plasma of the horse was collected from the jugular vein before euthanasia, and then placed in BD vacutainer EDTA-K2 (10.8 mg, 6 ml) and centrifuged at 3000 rpm for 10 minutes. Blood samples from normal horses and spontaneously infected horses were collected from the jugular vein after the ambulance and placed in BD vacutainer EDTA-K2 10.8 mg (6 ml) and centrifuged at 3000 rpm for 10 minutes.

(4) Protein Segmentation Way

The plasma proteins of horses were separated by size using SDS-PAGE electrophoresis, and the separated gels were divided into five equal sizes. 50 mM ammonium hydrogen carbonate (NH ₄ HCO ₃ ) containing 50% acetonitrile (ACN) was added to each gel, and vortexing was performed until the Coomassie Brilliant Blue (CBB) vortexing. It was then dehydrated with 100% acetonitrile and then vacuum dried for 20 minutes using a SpeedVac. To break down the gel, the gel pieces were reacted with 50 mM ammonium bicarbonate containing 10 mM DTT at 56 DEG C for 45 minutes. Thereafter, 50 mM ammonium hydrogencarbonate containing 55 mM iodoacetamide was added and cysteine alkylation was performed in the dark for 30 minutes. Finally, gel pieces were placed in 50 mM ammonium bicarbonate (pH 7.8) containing 12.5 ng / μl sequencing grade modified trypsin (Promega, Madison, Wis.) And allowed to incubate overnight at 37 ° C I have. After the gel was completely dissolved, a 50% acetonitrile solution containing 5% formic acid was added to extract a tryptic peptide, and the mixture was reacted at room temperature for 20 minutes. The supernatants were pooled, dried with SpeedVac, purified by resuspension in 0.1% formic acid, and concentrated using C18 ZipTips (Millipore, MA) prior to MS analysis.

(5) Nano-LC- ESI -MS / MS analysis

The purified trypsin peptide was loaded into a fused silica microinjection column (12 cm x 75 [mu] m) composed of C18 reverse phase resin (5 [mu] m, 200 A). Liquid chromatographic separation was carried out at a flow rate of 250 nL / min for 60 minutes under a linear gradient as follows: 3-40% solvent B (ACN with 0.1% formic acid), solvent A (0.1% formic acid Containing distilled water). The column was connected directly to an LTQ linear ion-trap mass spectrometer (Finnigan, CA) equipped with a nano-electrospray ion source. The electrospray voltage was 1.95 kV and the threshold value for switching from MS to MS / MS was 500. The normalized collision energy for MS / MS was 35% of the main radio frequency amplitude (RF) and the duration was 30 ms. All spectra were obtained in Data-dependent scan mode. Each full MS scan was due to five MS / MS scans, from the strongest peak to the fifth strongest peak of the entire MS. The number of repetitions of the peak for dynamic exclusion was 1, and the repetition time was 30s. The dynamic exclusion time was set to 180 s and the width of the exclusion mass was ± 1.5 Da.

(6) Database search and verification

The resulting LC-ESI-MS / MS fragmentation spectra were obtained from the BioWorkBrowser (www.ncbi.nlm.nih.gov), which contains the SEQUEST search engine from the National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov) ^TM (Version Rev. 3.3.1 SP1, Thermo Fisher Scientific Inc., CA). The search conditions were trypsin enzyme specificity, permissible level for 2 missed cleavage, and peptide tolerance.

2. Leprosy Identification of increased protein expression in plasma by induction

A comparative analysis was performed between the protein with increased expression level in plasma and the protein with increased expression level in plasma of natural endophthalmitis - induced horse than in normal horse due to artificial phyllotaxis induced by oligosaccharide administration. Among these, common proteins with an increase in plasma level and a change in drainage of 1.5 or more were selected by the artificial and natural infectious agent induction.

(One) Leprosy Plasma of induced horses Serpin Increased expression of A3-8-analog gene

Serpin A3-8-like (GI number: 338719856) protein has a 3.20-fold increase in the serum serine A3-8-analog protein level detected in plasma as compared to pre-triggered, Respectively.

Also, in comparative experiments with natural acfoderm-induced horses and normal control horses, the serpin A3-8-analog protein was detected only in the plasma of the natural acaflet-induced horses and not in the plasma of normal horses .

(2) Leprosy Plasma of induced horses C4b - confirmation of increased expression of binding protein alpha chain gene

The C4b-binding protein alpha chain (GI number: 149708028) protein increased the level of C4b-binding protein alpha chain detected in plasma by 1.62-fold as compared with that before induction,

In addition, in a comparison between the horse with naturally occurring phyllotaxis and the normal control group, the C4b-binding protein alpha chain protein was detected only in the plasma of the natural acaflet-induced horse and not in the plasma of the normal horse.

The selected proteins and their expression levels are shown in Table 1 below.

[Table 1] Myopia Plasma of artificially induced horses and two proteins with increased expression levels in plasma of naturally induced horses

3. Leprosy Identification of proteins with reduced expression levels in plasma by induction

(One) Leprosy Plasma keratin type II of induced horses Cytoskeleton Identification of the expression of the 1-analogue gene

A comparative analysis was performed between proteins with reduced expression levels in plasma and those with low levels of expression in plasma of horses induced by natural chlorophyll compared to normal horses due to the induction of artificial phyllotaxis by oligosaccharide administration. Among these, common proteins with reduced plasma expression and a change in drainage of 1.5 or more were selected by artificial and natural infectious agent induction.

The Keratin type II cytoskeletal 1-like (GI number: 194212030) protein decreased 2.74-fold in the keratin-type II cytoskeletal 1- .

In addition, in a comparison between the horse with the natural acafloid-induced and the normal control, the keratin type II cytoskeletal 1-analog protein was detected only in the plasma of normal horses and not in the plasma of natural clitoris-induced horses.

The selected keratin type II cytoskeletal 1-analog protein and its expression levels are shown in Table 2 below.

[Table 2] Plasma of Artificially Induced Horses and One Protein Commonly Expressed in Plasma of Spontaneously Induced Horses

<110> Seoul National University Industry Foundation <120> A composition and kit for detecting a laminitis, method for a laminitis and method for screening a therapeutic agent for a laminitis <130> PN111854 <160> 3 <170> Kopatentin 2.0 <210> 1 <211> 473 <212> PRT <213> Equus ferus <400> 1 Met Ser Lys Leu Val Ala Val Gly Lys Gly Pro Arg Ala Leu Thr Leu 1 5 10 15 Trp Ala Tyr Ser Arg Gly Arg Phe Arg Arg Gly Gly Arg Arg Gln His 20 25 30 Trp Ser Arg Gln Ala Ser Arg Ala Gln Leu Ser Glu Ala Glu Leu Arg 35 40 45 Ala Glu Arg Met Ser Leu Leu Ala Leu Gly Leu Leu Val Ala Gly 50 55 60 Leu Cys Pro Thr Val His Cys Leu Pro Gly Asp Met Pro Asp Ser Gly 65 70 75 80 Asn Val Ala Gln Glu Asp Gln Pro Asn Arg Thr Ser Val Asp Asn Leu 85 90 95 Arg Leu Thr Thr Ser Asn Thr Asp Phe Ala Phe His Leu Tyr Lys Leu 100 105 110 Leu Ala Gln Thr Pro Asn Glu Asn Val Ile Phe Ser Pro Leu Ser 115 120 125 Ile Ser Ile Ala Leu Ala Phe Leu Ser Leu Gly Ala Arg Asn Thr Thr 130 135 140 Leu Thr Glu Ile Leu Glu Gly Leu Lys Phe Asn Leu Thr Glu Thr Pro 145 150 155 160 Glu Thr Glu Ile His Gln Gly Phe Gln His Leu Leu His Thr Leu Ser 165 170 175 Gln Pro Asn Pro Gln Gln Gln Leu Ser Val Gly Asn Ala Met Phe Leu 180 185 190 Lys Glu Glu Leu Glu Leu Leu Asp Lys Phe Arg Glu Asp Ala Lys Ala 195 200 205 Leu Tyr Ala Ser Glu Ala Phe Pro Thr Asp Phe Lys Asp Pro Ala Ala 210 215 220 Ala Glu Lys Leu Ile Asn Asp Tyr Val Glu Lys Lys Thr Glu Gly Lys 225 230 235 240 Ile Val His Leu Val Gly Gly Leu Cys Lys Asn Thr Met Met Val Leu 245 250 255 Val Asn Tyr Ile Leu Leu Lys Ala Lys Trp Lys Thr Pro Phe Asp Pro 260 265 270 Gly Asp Thr Tyr Glu Ser Ala Phe His Val Ser Arg Ser Ser Arg Ser Val 275 280 285 Gln Val Pro Lys Met Ser Phe Glu Glu Arg Met Val Pro Phe Phe Arg 290 295 300 Asp Glu Glu Leu Ala Cys Thr Val Val Glu Leu Gln Tyr Ile Ser Asp 305 310 315 320 Asp Ser Ala Leu Phe Ile Leu Pro Asp Glu Gly Gln Leu Gly Ala Val 325 330 335 Glu Ala Leu Leu Pro Glu Thr Leu Arg Arg Trp Arg Ala Ser Leu 340 345 350 Arg Met Arg Trp Ile Asp Glu Leu Tyr Leu Pro Lys Phe Ser Ile Ser 355 360 365 Ser Asn Tyr Glu Leu Glu Thr Ile Leu Thr Gln Leu Gly Ile Glu Lys 370 375 380 Val Phe Thr Thr Lys Ala Asp Leu Ser Gly Val Thr Gly Thr Pro Asn 385 390 395 400 Leu Tyr Val Thr Gln Val Val His Ser Thr Val Leu Asp Val Ala Glu 405 410 415 Glu Gly Thr Glu Ala Ala Ala Thr Gly Ile Asn Ile Ala Phe Ser 420 425 430 Ser Gly Val Met Asn Pro Leu Ile Val Asp Phe Asn Arg Pro Phe Leu 435 440 445 Leu Phe Ile Ile Ser Lys Asp Thr Gln Ser Ile Leu Phe Gly Gly Lys 450 455 460 Val Val Asp Pro Ser Gln Ala Pro His 465 470 <210> 2 <211> 595 <212> PRT <213> Equus ferus <400> 2 Met His Pro Pro Arg Ala Pro Asn Gly Thr Leu His Arg Lys Trp Lys 1 5 10 15 Thr Ala Ala Trp Pro Phe Ser Arg Leu Trp Arg Leu Ser Asp Pro Thr 20 25 30 Leu Phe Gln Val Thr Leu Val Ile Ala Leu Leu Ala Thr Val Leu Gly 35 40 45 Asp Cys Gly Pro Pro Pro Asn Leu Leu Phe Ala Ser Pro Ile Asn Glu 50 55 60 Leu Asn Glu Thr Asp Phe Glu Ala Gly Thr Ile Leu Lys Tyr Asn Cys 65 70 75 80 Arg Pro Gly Tyr Ser Lys Thr Ser Ser Lys Asn Ser Leu Thr Cys Gln 85 90 95 His Gly Gly Val Trp Lys Tyr Thr Pro Phe Cys Thr Lys Lys Arg Cys 100 105 110 Arg Asn Pro Gly Glu Leu His Asn Gly Gln Ile Ser Lys Thr Asp Leu 115 120 125 Ser Phe Gly Ser Arg Ile Glu Phe Ser Cys Ser Asp Gly Tyr Ile Leu 130 135 140 Ile Gly Ser Thr Thr Ser Tyr Cys Asp Ile Gln Asp Lys Gly Val Asp 145 150 155 160 Trp Ser Asp Pro Leu Pro Val Cys Val Ile Ala Lys Cys Glu Ala Pro 165 170 175 Pro Ala Ile Ser Asn Gly Lys His Ser Gly Gly Asp Glu Asp Val Tyr 180 185 190 Thr Tyr Gly Ser Ser Val Thr Tyr Ser Cys Asp Pro His Phe Ser Met 195 200 205 Ile Gly Lys Ala Ser Ile Ser Cys Thr Val Glu Asn Lys Thr Ile Gly 210 215 220 Val Trp Ser Pro Ser Pro Thr Cys Lys Tyr Val Val Cys His Lys 225 230 235 240 Pro Gln Val Pro Asn Gly Ile Phe Val Ser Gly Phe Gly Pro Leu Tyr 245 250 255 Thr Tyr Lys Asp Ala Ile Val Leu Asp Cys Lys Lys Gly Tyr Val Leu 260 265 270 Thr Gly Ser Ser Leu Ile His Cys Glu Ala Asp Asn Asn Trp Asp Pro 275 280 285 Pro Pro Pro Val Cys Glu Leu Asn Gly Cys Thr Asn Leu Pro Asp Ile 290 295 300 Pro Asn Ala Phe Trp Glu Arg Tyr Arg Tyr Gln Arg Pro Thr Lys Glu 305 310 315 320 Asp Val Tyr Asn Val Gly Thr Val Leu Lys Tyr His Cys Leu Thr Gly 325 330 335 Tyr Lys Pro Ala Ser Asp Lys Pro Thr Ala Val Ile Cys Gln Glu Asp 340 345 350 Phe Arg Trp Thr Pro Tyr Thr Glu Cys Glu Glu Val Cys Cys Pro Val 355 360 365 Pro Glu Leu Lys Asn Gly Glu Ile Phe Ser Gln Arg Arg Arg Ala Ser 370 375 380 Thr Ser Ser Cys Val Tyr Ser Ser Gly Glu Lys Ile Thr Ser Ser Cys 385 390 395 400 Phe Lys Lys Lys Glu Phe Ser Ala Thr Cys Leu Lys Asp Gly Thr Trp 405 410 415 His Pro Lys Thr Pro Val Cys Asp Asp Cys Ile Phe Pro Ala Thr Ile 420 425 430 Asp His Gly His Tyr Lys Glu Val Asn Gly Phe Trp Thr Asn Glu Val 435 440 445 Leu Tyr Glu Cys Asp Glu Gly Tyr Ser Leu Val Gly Gln Ala Arg Leu 450 455 460 Ser Cys Asn Ser Ser Gly Trp Ser Ser Pro Ala Pro Gln Cys Lys Ala 465 470 475 480 Leu Cys Pro Lys Pro Gln Ile Glu His Gly Arg Leu Ser Val Val Lys 485 490 495 Asp Gln Tyr Ile Thr Pro Glu Asn Val Thr Ile Gln Cys Asp Pro His 500 505 510 Tyr Arg Leu Val Gly Leu Gln Ser Ile Thr Cys Ser Glu Asn Ser Thr 515 520 525 Trp Tyr Pro Glu Val Pro Met Cys Glu Trp Glu Ile Ala Glu Gly Cys 530 535 540 Glu Gln Val Leu Ala Gly Arg Lys Ile Met Gln Cys Leu Pro Lys Pro 545 550 555 560 Glu Asp Val Arg Thr Ala Leu Glu Leu Tyr Lys Leu Ser Leu Glu Ile 565 570 575 Lys Gln Leu Glu Lys Lys Leu Glu Lys Glu Glu Lys Cys Thr Pro Glu 580 585 590 Val Gln Glu 595 <210> 3 <211> 618 <212> PRT <213> Equus ferus <400> 3 Met Ser Cys Arg Ile Ser Ser Arg Ser Ser Arg Gly Arg Gly Gly Gly Gly 1 5 10 15 Gly Gly Gly Phe Arg Gly Phe Ser Ser Ser Ser Ala Val Gly Ser Gly 20 25 30 Gly Ser Arg Arg Ser Thr Ser Ser Phe Ser Cys Leu Ser Arg His Gly 35 40 45 Gly Gly Gly Arg Gly Val Gly Gly Gly Gly Gly Phe Gly Ser Arg Ser Leu 50 55 60 Val Gly Leu Gly Gly Thr Arg Ser Ile Ser Ile Ser Val Ala Gly Gly 65 70 75 80 Gly Gly Ser Phe Gly Ser Gly Gly Gly Phe Gly Gly Arg Gly Gly Gly 85 90 95 Phe Gly Gly Gly Ile Gly Phe Gly Gly Gly Ser Gly Phe Gly Gly Gly 100 105 110 Ser Gly Phe Gly Gly Gly Gly Ply Gly Gly Gly Gly Gly Phe Gly Gly Gly 115 120 125 Arg Phe Gly Gly Gly Ser Gly Phe Gly Gly Phe Gly Gly Pro Gly Gly 130 135 140 Phe Gly Pro Gly Gly Phe Pro Gly Gly Gly Ile His Glu Val Ser Ile 145 150 155 160 Asn Gln Ser Leu Leu Gln Pro Leu Asn Val Gly Val Asp Pro Glu Ile 165 170 175 Gln Asn Val Lys Ala Gln Glu Arg Glu Gln Ile Lys Thr Leu Asn Asn 180 185 190 Lys Phe Ala Ser Phe Ile Asp Lys Val Arg Phe Leu Glu Gln Gln Asn 195 200 205 Gln Val Leu Gln Thr Lys Trp Glu Leu Leu Gln Gln Val His Val Gly 210 215 220 Thr Arg Thr Ser Asn Leu Glu Pro Ile Phe Gln Ala Phe Ile Ala Gln 225 230 235 240 Leu Lys Arg Gln Val Asp Thr Leu Cys Ala Glu Arg Thr Ser Gln Asp 245 250 255 Ser Glu Leu Asn Ser Met Gln Asp Leu Val Glu Asp Phe Lys Lys Lys 260 265 270 Tyr Glu Asp Glu Ile Asn Arg Arg Thr Ala Ala Glu Asn Asp Phe Val 275 280 285 Thr Leu Lys Lys Asp Val Asp Asn Ala Tyr Lys Ile Lys Val Asp Leu 290 295 300 Gln Ala Lys Val Asp Val Leu Thr Gln Glu Leu Glu Phe Leu Arg Ile 305 310 315 320 Leu Tyr Asp Ala Glu Leu Ser Gln Met Gln Gln Ser Ile Ser Asp Thr 325 330 335 Ser Val Val Leu Ser Met Asp Asn Ser Arg His Leu Asp Leu Asn Ser 340 345 350 Ile Ile Ala Glu Val Ala Gln Tyr Glu Glu Ile Ala Gln Lys Ser 355 360 365 Lys Ala Glu Ala Glu Ala Leu Tyr Gln Ser Lys Tyr Glu Glu Leu Gln 370 375 380 Ile Thr Ala Gly Ser Gly Asp Ser Leu Lys Ser Thr Lys Met Glu 385 390 395 400 Ile Ser Glu Leu Asn Arg Val Ile Gln Arg Leu Arg Ser Glu Ile Asp 405 410 415 Ser Val Lys Lys Gln Ile Ala Ala Leu Gln Gln Ser Ile Ser Glu Ala 420 425 430 Glu Gln Arg Gly Glu Asn Ala Leu Lys Asp Ala Gln Asn Lys Leu Asn 435 440 445 Glu Leu Glu Ala Ala Leu Gln Arg Ala Lys Glu Asp Leu Ala Arg Leu 450 455 460 Leu Arg Asp Tyr Gln Glu Leu Met Ser Thr Lys Leu Ala Leu Asp Val 465 470 475 480 Glu Ile Ala Thr Tyr Arg Thr Leu Leu Glu Gly Glu Glu Gly Arg Met 485 490 495 Ser Gly Glu Cys Ala Pro Asn Val Ser Val Ser Val Ser Thr Ser His 500 505 510 Thr Ser Ile Ser Gly Gly Gly Val Gly Gly Gly Gly Gly Phe Ser Ser 515 520 525 Gly Gly Gly Gly Ser Tyr Met Ser Gly Gly Gly Ser Ser Ser Ser Gly 530 535 540 Gly Gly Gly Gly Gly Phe Gly Ser Gly Gly Ser Ser Arg Gly His Arg 545 550 555 560 Gly Gly Ser Gly Gly Gly Gly Ser Gly Ser Tyr Gly Val Arg Ile Ser Gly 565 570 575 Gly Gly Gly Ser Gly Gly Ser Phe Ser Ser Ser Gly Gly Arg Gly Val 580 585 590 Ser Ser Gly Ser Ser Ser Ser Ser Ser Ser Ser Ser Val 595 600 605 Val Ser Ser Ser Tyr Ser Gln Gly Ser Arg 610 615

Claims

Serpin A3-8-like gene, C4b-binding protein alpha chain gene or Keratin type II cytoskeletal 1-like gene, A composition for the diagnosis of horses' dermatophytes, comprising an agent for measuring a protein expression level of a gene.

delete

The composition of claim 1, wherein the agent comprises a substance that specifically binds to a serine A3-8-analog, a C4b-binding protein alpha chain, or a keratin type II cytoskeletal 1- .

4. The method of claim 3, wherein the substance is selected from the group consisting of a primer, a probe, a nucleotide, an antibody, or an antigen-binding fragment thereof, specifically binding to a serine A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1- , A ligand, a receptor, an agonist or an antagonist, a protein, or a combination thereof.

A kit for diagnosing acute myeloid leukemia comprising an agent for measuring a protein expression level of a serine A3-8-analog gene, a C4b-binding protein alpha chain gene or a keratin type II cytoskeleton 1-analog gene.

Contacting a sample separated from horses with a substance that specifically binds to a serpin A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeletal 1-analog protein to form a complex thereof;
Determining the level of protein expression of the serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene in the sample by measuring the level of the complex;
The protein expression levels of the measured serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene were measured by using serpin A3-8-analog, C4b- With a protein expression level of a protein alpha chain or a keratin type II cytoskeletal 1-analog; And
When the protein expression level of the serine A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeletal 1-analog gene of the horse is changed compared to the protein expression level of the normal control, The method comprising the steps of:

[Claim 6] The method according to claim 6, wherein when the protein expression level of the serine A3-8-analog gene, C4b-binding protein alpha chain gene of the horse is higher than that of a normal control protein, Lt; RTI ID = 0.0 > fibroblast < / RTI >

7. The method of claim 6, wherein the step of determining is that if the protein expression level of the horse's keratin type II cytoskeletal 1- analogue gene is low compared to the level of protein expression of a normal control, How to include.

delete

7. The method of claim 6, wherein the step of measuring the level of expression comprises the steps of Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion Immunoprecipitation Assay, Complement Fixation Assay, FACS, Mass Spectrometry, Magnetized Bead-antibody Immunoassay, Protein Chip, Immunoprecipitation Assay, Immunoprecipitation Assay, Or a combination thereof.

Administering a candidate agent to the horse;
Contacting a sample separated from said horse with a substance that specifically binds to a serpin A3-8-analog, a C4b-binding protein alpha chain or a keratin type II cytoskeleton 1-analog protein to form a complex thereof;
Determining the level of protein expression of the serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene in the sample by measuring the level of the complex;
The protein expression level of the measured serpin A3-8-analog gene, C4b-binding protein alpha chain gene or keratin type II cytoskeleton 1-analog gene was measured using a serine A3-8-analog gene, C4b- With a protein expression level of a binding protein alpha chain gene or keratin type II cytoskeletal 1-analog gene; And
When the protein expression level of the horse serine A3-8-analog gene, the C4b-binding protein alpha chain gene or the keratin type II cytoskeleton 1-analog gene is changed as compared to the protein expression level of the normal control, Lt; RTI ID = 0.0 > of < / RTI > chlorophyll.