KR102114443B1 - Marker for diagnosing Charcot-Marie-Tooth and use thereof - Google Patents

Abstract

 Provided are compositions for diagnosing the risk of developing Sharko-Marie-Tus and kits comprising the same. In addition, it provides a method for providing information for predicting the risk of developing Charco-Marie-Tooth using the same. The diagnosis can accurately diagnose the risk of developing Charco-Marie-Tooth early through genetic testing, thereby maximizing the therapeutic effect.

Description

Marker for diagnosing Charcot-Marie-Tooth and use thereof

It relates to compositions, kits and uses thereof for diagnosing the risk of developing Sharko-Marie-Tus.

Inherited neuropathy (IN) is a congenital disease that has a family history and indicates dysfunction of the motor and sensory nervous systems by mutation of genes. It is largely divided into hereditary motor and sensory neuropathy (HMSN), hereditary motor neuropathy (HMN), and hereditary sensory neuropathy (HSN). Of these, HMSN accounts for the majority, which is known as Charcot-Marie-Tooth disease (CMT). CMT disease is characterized by muscle weakness and atrophy that progresses slowly in the lower extremity fibula muscle, and is characterized by arereflexia, distal sensory loss, pes cavus, and deafness. It is also indicated. The outbreak usually occurs before and after the teens, but rarely after the thirties. However, the clinical phenotype and age of onset of CMT disease are very heterogeneous and neural biopsy for confirmation is not easy, making accurate diagnosis and treatment difficult (Berger P., Young P., Suter U. Neurogenet. 4: 1-15 (2002)).

Recently, with the development of human genome research and bioinformatics, research into separating genes causing genetic diseases and revealing molecular biological mechanisms has been very active worldwide. Genetic cause analysis of genetic neuropathy, known to be caused by genetic factors rather than environmental factors, has also been carried out, and so far, most mutations except 17p11.2-p12 1.4-1.5Mb overlap/deletion It was found to be due to overlap/deletion of SNP and short nucleotide sequence.

According to the accurate genetic diagnosis of CMT, it is possible to suppress the outbreak and tailor treatment suitable for the genetic cause, but the development of an effective CMT diagnosis method is still insufficient. For personalized treatment of genetic neurological disorders with very heterogeneous characteristics, it is first necessary to establish an accurate genetic diagnosis method. As a result, the present inventors tried to find out the cause of the onset of CMT, and as a result of researching in detail, identified a new mutant gene that has not been previously reported, and completed the present invention by confirming that the mutant gene can be usefully used for diagnosis of CMT. Did.

 One aspect provides a composition for diagnosing the risk of developing Sharko-Marie-Tus and a kit comprising the same.

Another aspect provides a method for providing information to predict an individual's risk of developing Sharko-Marie-Tus.

One aspect is a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, the polynucleotide comprising 10 or more contiguous nucleotides selected from the polynucleotide and comprising a mutation position of the polynucleotide, wherein The position of the mutation is the risk of developing Sharko-Marie-Tooth in an individual who is the 47th nucleotide, 112th nucleotide, 139th nucleotide, 157th nucleotide, 414th nucleotide, 328th nucleotide, or combination thereof from the 5'end of SEQ ID NO: 1 It provides a composition for diagnosing.

CMT is a hereditary neurological disease that exhibits symptoms of distal muscle weakness accompanied by loss of sensory function, and may be classified as CMTX, CMT1, CMT2, CMT4, or IntCMT depending on the phenotype or genotype. CMTX is related to the X chromosome, and refers to a peripheral sensory motor multiple neurological disease in which the limb muscles associated with the feet, legs, and hands weaken and atrophy, and progress slowly and progressively. Includes. The composition for diagnosing the risk of developing Sharko-Marie-Tus may be for diagnosing the risk of CMTX. The CMTX may be CMTX1.

"Risk-Marie-Tooth development risk" may be a relative risk of Sharko-Marie-Tus development risk. For example, the risk may indicate whether the probability of onset is increased or decreased compared to a healthy normal group. In addition, the risk may indicate whether an individual with a specific allele or genotype has an increased or decreased probability of developing Sharko-Marie-Tus compared to an individual with a specific allele or genotype.

The subject refers to a subject for predicting the risk of developing Sharko-Marie-Tus. The subject may include vertebrates, mammals, or humans ( Homo sapiens ). For example, the human may be Korean.

"Polynucleotide" can be DNA or RNA. The polynucleotide may also be in single-stranded or double-stranded form. The polynucleotide may also be composed of natural nucleotides, as well as natural nucleotides, analogues of natural nucleotides, sugars, bases or phosphate sites of natural nucleotides, as long as they have properties that can be hybridized by hydrogen bonding to complementary nucleotides. And nucleotides selected from the group consisting of these nucleotides (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews, 90:543-584 (1990)). .

The polynucleotide represents a single nucleotide polymorphism at the mutation site. Therefore, when one single-stranded polynucleotide is associated with the risk of developing Sharko-Marie-Tus, it can be determined that the polynucleotide complementary to the single-stranded polynucleotide is naturally associated with the risk of developing Sharko-Marie-Tus. have. Thus, a composition according to one aspect comprises a single-stranded polynucleotide associated with a risk of developing Sharko-Marie-Tus having one specific sequence and a polynucleotide having a sequence complementary thereto.

For example, the polynucleotide of SEQ ID NO: 1 is the 47th (SNP position) nucleotide "A or G". In this case, the composition comprises the "A or G" nucleotide of the 47th (SNP position) and 10 or more consecutive nucleotides selected from the polynucleotide of SEQ ID NO: 1, as well as the position corresponding to the 47th (SNP position). Complementary single-stranded polynucleotides having “T or C” nucleotides. That is, the composition can detect the 47th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect a c.47A>G mutation in the polynucleotide of SEQ ID NO: 1.

The polynucleotide of SEQ ID NO: 1 is the 112th (SNP position) nucleotide "G or T". In this case, the composition comprises the "G or T" nucleotide of the 112th (SNP position) and 10 or more consecutive nucleotides selected from the polynucleotide of SEQ ID NO: 1, as well as the position corresponding to the 112th (SNP position). Complementary single-stranded polynucleotides having “C or A” nucleotides. That is, the composition can detect the 112 th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect c.112G>T mutation in the polynucleotide of SEQ ID NO: 1.

The polynucleotide of SEQ ID NO: 1 is the 139th (SNP position) nucleotide "G or A". In this case, the composition comprises a "G or A" nucleotide at position 139 (SNP position) and at least 10 contiguous nucleotides selected from polynucleotides of SEQ ID NO: 1, as well as at positions corresponding to position 139 (SNP position). And complementary single-stranded polynucleotides having “C or T” nucleotides. That is, the composition can detect the 139th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect c.139G>A mutation in the polynucleotide of SEQ ID NO: 1.

The polynucleotide of SEQ ID NO: 1 is the 157th (SNP position) nucleotide "T or C". In this case, the composition comprises a "T or C" nucleotide at the 157th (SNP position) and 10 or more consecutive nucleotides selected from the polynucleotide of SEQ ID NO: 1, as well as a position corresponding to the 157th (SNP position). Complementary single-stranded polynucleotides having “A or G” nucleotides. That is, the composition can detect the 157th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect c.157T>C mutation in the polynucleotide of SEQ ID NO: 1.

The polynucleotide of SEQ ID NO: 1 is 414 th (SNP position) nucleotide "C or G". In this case, the composition comprises a "C or G" nucleotide at position 414 (SNP position) and at least 10 consecutive nucleotides selected from the polynucleotides of SEQ ID NO: 1, as well as at positions corresponding to position 414 (SNP position). Complementary single-stranded polynucleotides having “G or C” nucleotides. That is, the composition can detect the 414 th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect c.414C>G variation in the polynucleotide of SEQ ID NO: 1.

The polynucleotide of SEQ ID NO: 1 is 328th (SNP position) nucleotide "G or A". In this case, the composition comprises the "G or A" nucleotide of the 328th (SNP position) and 10 or more consecutive nucleotides selected from the polynucleotide of SEQ ID NO: 1, as well as the position corresponding to the 328th (SNP position) And complementary single-stranded polynucleotides having “C or T” nucleotides. That is, the composition can detect the 328th nucleotide from the 5'end of SEQ ID NO: 1, which is the SNP position in the polynucleotide of SEQ ID NO: 1. Alternatively, the composition can detect c.328G>A mutation in the polynucleotide of SEQ ID NO: 1.

The nucleotide at the variation position of the nucleotide sequence of SEQ ID NO: 1 may be used as a marker for diagnosing or predicting the risk of developing Charco-Marie-Tus.

“Variation” refers to alteration of one or more nucleotides in the genome.

“Variation location” means the location of the mutation on the genome.

Variations may include substitution of one or more nucleotides, insertion, deletion (insertion, deletion also referred to as Insertion, Deletion:'InDel'), and the like. Substitution means a change in which a nucleotide is replaced with another nucleotide. Insertion refers to a change in which one or more nucleotides are added. Deletion refers to a change in which one or more nucleotides are removed.

Single nucleotide polymorphism (single nucleotide polymorphism: SNP, referred to as SNP) is used in the sense commonly known in the art. SNPs can represent single nucleotide polymorphisms present in the genome within a population. The SNP may be a genotype at the SNP position. The SNP may be that the frequency of minority alleles of the SNP in the population is 1% or more. SNPs can be mixed with single nucleotide variants or single nucleotide variants (SNV, hereinafter referred to as SNV).

Base A means adenine, base G means guanine, base C means cytosine, and base T means thymine.

The polynucleotide may be 10 to 200 nucleotides in length (nucleotide: nt). For example, the polynucleotide may have a length of 10 to 150 nt, 10 to 100 nt, or 15 to 100 nt.

The polynucleotide may be singular or plural, or 1 pair or plural pairs.

The polynucleotide may be a primer, probe or antisense nucleic acid. "Primer" refers to a single-stranded polynucleotide capable of acting as a starting point in the polymerization reaction of a nucleotide by a polymerase. For example, the primer can be a single strand capable of acting as a starting point for template-directed DNA synthesis under suitable conditions (i.e. the presence of four different nucleoside triphosphates and polymerases) in a suitable temperature and a suitable buffer. It may be a polynucleotide of. The suitable length of the primer can vary depending on various factors, such as temperature and the use of the primer. The primer may be 15 to 30 nt in length. Short primer molecules generally require lower temperatures to form sufficiently stable hybrid complexes with the template.

The sequence of the primer need not have a completely complementary sequence with some sequences of the template, and it is sufficient to have a complementarity within a range capable of hybridizing with the template and working with the primer. Therefore, the primer includes not only the polynucleotide itself, but also a sequence that hybridizes specifically to the polynucleotide described above, and can also serve as a starting point in a polymerization reaction. For example, it may be a sequence perfectly complementary to the polynucleotide of SEQ ID NO: 1, as well as a sequence having hybridization to this sequence and having complementarity within a range capable of acting as a primer. The design of the primer can be easily performed by a person skilled in the art by referring to the sequence of the target nucleic acid to be amplified. For example, it may be designed using a commercially available primer design program. Examples of such commercially available primer design programs include the PRIMER 3 program.

When the polynucleotide is used as a PCR primer, in addition to the polynucleotide, it may include a primer that specifically binds its complementary strand.

“Probe” refers to a polynucleotide that specifically binds a specific target sequence. The polynucleotide may be DNA or RNA. The polynucleotide may be in the form of a single strand. The polynucleotide may also be composed of natural nucleotides, as well as natural nucleotides, analogues of natural nucleotides, sugars, bases or phosphate sites of natural nucleotides, as long as they have properties that can be hybridized by hydrogen bonding to complementary nucleotides. It may include a nucleotide selected from the group consisting of nucleotides and combinations thereof. The probe may have a length of 5 to 100nt, 10 to 90nt, 15 to 80nt, 20 to 70nt, or 30 to 50nt. The polynucleotide includes PNA. In addition, the polynucleotide may be attached with a detectable label (eg, Cy3, Cy5 fluorescent substance), for example, for ease of detection of the polynucleotide or a complex to which it is bound in an analytical reaction, eg, 3' It may be attached to the terminal or 5'end.

The probe may be a sequence that is completely complementary to the target sequence including a mutation site. In addition, the probe may have a nucleotide sequence that is substantially complementary within a range that does not interfere with specific hybridization to the target sequence including the mutation site. In addition, the probe may be one having modified nucleotides within a range that does not impair specific hybridization to the target sequence including the mutation site. Examples of the probes include a perfect match probe consisting of a sequence perfectly complementary to a polynucleotide containing a mutation site and a polynucleotide comprising a mutation position, completely complementary to all sequences except the mutation position. It may be selected from the group consisting of probes having a positive sequence.

"Antisense nucleic acid" means a nucleic acid-based molecule that has a nucleotide sequence complementary to a target nucleic acid and is capable of forming a dimer therewith. The antisense nucleic acid may be the polynucleotide or a fragment thereof, or complementary thereto. The antisense nucleic acid may have a length of 10 nt or more, more specifically 10 to 200 nt, 10 to 150 nt, or 10 to 100 nt, but an appropriate length may be selected to increase detection specificity.

The primer, probe, or antisense nucleic acid can be used to amplify or confirm the presence of a nucleotide sequence having a specific allele at the mutation site.

The composition may be a polynucleotide composed of the nucleotide sequence of SEQ ID NO: 1 or a complementary polynucleotide thereof, and may include a combination of mutation positions of the polynucleotide.

The polynucleotide may be labeled with a detectable label. The detectable label is a label material capable of generating a detectable signal, and may be a label material capable of generating a detectable signal including a fluorescent material, for example, materials such as Cy3 and Cy5. The detectable label can confirm the hybridization result of the nucleic acid.

The polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 may be a polynucleotide encoding the GJB1 protein. The GJB1 (Gap junction beta-1 protein) gene, also called connexin 32 (Cx32), is a member of the gap junctional connectin family and encodes a transmembrane protein that controls signal transduction through the cell membrane in the peripheral nervous system. As a gene, it may be a gene registered with NCBI Accession number NM_000166.5. The gene may be a polynucleotide having a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 2. The GJB1 protein may be a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 1, a polypeptide comprising the amino acid sequence of SEQ ID NO: 2, or a protein registered with NCBI Accession number NP_000157.1.

Those skilled in the art will be able to easily identify the location of the variation, nucleotide sequence and amino acid sequence using the registration number. The specific sequence corresponding to the number registered in the UCSC genome browser or GenBank may change somewhat over time. It will be apparent to those skilled in the art that the scope of the present invention also affects the altered sequence.

The composition may be a solid composition or a liquid composition. The liquid composition may include the polynucleotide in a suitable liquid medium. The liquid medium may be any used in the art as a liquid medium for polynucleotides. In addition, the liquid medium may be appropriately selected according to the purpose of using the composition. For example, the liquid medium may be selected from the group consisting of water, PCR buffer and hybridization buffer.

Another aspect provides a microarray for diagnosing the risk of developing Sharko-Marie-Tooth comprising the composition. The polynucleotide, composition, individual, and Sharco-Marie-Tus are as described above. "Microarray" means that the polynucleotide is immobilized at a high density in a distinct region of the substrate surface. In the microarray, the region may be arranged on a substrate at a density of 400/cm ² or more, 10 ³ /cm ² , or 10 ⁴ /cm ² , for example.

Another aspect provides a kit for diagnosing the risk of developing Sharko-Marie-Tooth comprising the composition. The polynucleotide, composition, individual, and Sharco-Marie-Tus are as described above.

The kit may be a kit for specifically amplifying a target sequence and diagnosing an individual's risk of developing Sharko-Marie-Tooth through the presence or absence of an amplification product. In this case, the kit may include the polynucleotide as a primer and a reagent necessary for amplification. The amplification reagent may include, for example, dNTP, polymerase, and a suitable buffer. In the primer, for example, the nucleotide sequence corresponding to the mutation site forms the 3'terminal nucleotide of the primer, and the 3'terminal nucleotide is complementary to the nucleotide of the mutation position (specific primer) or not complementary. (Non-specific primers). The non-specific primer may include a sequence that is not complementary to the 3'terminal nucleotide as well as other sites. The kit may also include instructions for use. The instructions for use, for example, when the target sequence is amplified in the amplification reaction using the specific primer and the target sequence is not amplified in the amplification reaction using the non-specific primer, Sharco-Marie-Tooth among samples used for amplification. It may include a description of the outcome determination, including a description of determining that a target sequence associated with is present and determining the risk of developing an individual's Sharko-Marie-Tus from the results.

The kit may be a kit for hybridizing the polynucleotide or a probe derived therefrom with a nucleic acid in a sample, and diagnosing an individual's risk of developing Charco-Marie-Tus from the hybridization result. In this case, the kit may include the probe and reagents necessary for hybridization. Reagents required for hybridization may include, for example, a hybridization buffer. The nucleic acid may be amplified or unamplified. Therefore, the kit may further include reagents necessary for amplification of nucleic acids. The nucleic acid can be labeled with a detectable label. The kit may include a probe that is completely complementary to the target sequence (perferct match probe) or, in the target sequence, a mismatch probe that is complementary to all regions except the mutation site. The kit may also include instructions for use. The instructions for use include, for example, a sequence associated with Sharco-Marie-Tooth in a sample when a target sequence is detected in a hybridization reaction using the completely complementary probe, and a target sequence is not detected in a hybridization reaction using the mismatch probe. It may include a description of the outcome decision, including a description of determining that it exists and determining the risk of developing the individual's Sharko-Marie-Tus from the results.

Another aspect of the polypeptide consisting of the amino acid sequence of SEQ ID NO: 2, a polypeptide for specifically detecting a polypeptide comprising the amino acid mutation position of the polypeptide, wherein the amino acid mutation position is the 16th from the N-terminal of SEQ ID NO: 2 Provided are compositions for diagnosing the risk of developing Sharko-Marie-Tooth in an individual who is an amino acid, 38th amino acid, 47th amino acid, 53rd amino acid, 138th amino acid, 110th amino acid, or a combination thereof.

The polypeptide represents a single nucleotide polymorphism at the mutation position, thereby indicating amino acid substitution.

For example, the polypeptide of SEQ ID NO: 2 is the 16th (amino acid substitution position) amino acid "H or R". That is, the composition can detect the 16th amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition may detect a p.H16R mutation in the polypeptide of SEQ ID NO: 6.

The polypeptide of SEQ ID NO: 2 is the 38th (amino acid substitution position) amino acid "V or L". That is, the composition can detect the 38th amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition may detect a p.V38L mutation in the polypeptide of SEQ ID NO: 2.

The polypeptide of SEQ ID NO: 2 is the 47th (amino acid substitution position) amino acid "E or K". That is, the composition can detect the 47th amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition can detect a p.E47K mutation in the polypeptide of SEQ ID NO: 2.

The polypeptide of SEQ ID NO: 2 is the 53rd (amino acid substitution position) amino acid "C or R". That is, the composition can detect the 53rd amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition may detect a p.C53R mutation in the polypeptide of SEQ ID NO: 2.

The polypeptide of SEQ ID NO: 2 is the 138th (amino acid substitution position) amino acid "S or R". That is, the composition can detect the 138th amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition may detect a p.S138R mutation in the polypeptide of SEQ ID NO: 2.

The polypeptide of SEQ ID NO: 2 is the 110th (amino acid substitution position) amino acid "G or S". That is, the composition can detect the 110th amino acid from the N-terminal of SEQ ID NO: 2, which is the amino acid substitution position in the polypeptide of SEQ ID NO: 2. Alternatively, the composition may detect a p.G110S mutation in the polypeptide of SEQ ID NO: 2.

The polypeptide comprising the amino acid sequence of SEQ ID NO: 2 may be a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 1, a protein registered with NCBI Accession number NP_000157.1, or a human GJB1 protein.

The GJB1 protein is a transmembrane protein located across EC (extracellular, extracellular), TM (transmembrane), and IC (intracellular, intracellular). The p.H16R mutation may be located in the IC domain of the GJB1 protein. The p.V38L mutation may be located in the TM domain of the GJB1 protein. The p.E47K mutation may be located in the EC domain of the GJB1 protein. The p.C53R mutation may be located in the EC domain of the GJB1 protein. The p.S138R mutation may be located in the TM domain of the GJB1 protein. The p.G110S mutation may be located in the IC domain of the GJB1 protein.

The amino acid at the amino acid variation position of the amino acid sequence of SEQ ID NO: 2 may be used as a marker for diagnosing or predicting the risk of developing Sharko-Marie-Tus.

Amino acid variation means that the amino acid sequence is changed by changing one or more nucleotides encoding the amino acid. Amino acid mutations include missense mutations, latent mutations, nonsense mutations, neutral mutations, intron mutations, and frame shift mutations. Frame shift) and the like. A missense variation refers to a change in which nucleotides are changed to encode other amino acids. Potential variation refers to a variation in which the nucleotide is encoded, but the amino acid being encoded is the same. Nonsense variation means a change that no longer produces amino acids by changing the nucleotide to become a termination codon. Neutral variation refers to a change in nucleotides that has been changed to encode another amino acid, but the same nature as the originally designated amino acid. Variation in the intron refers to the change in the non-encoding intron region. The lattice shift mutation refers to a change in which amino acids to be translated are changed by shifting the decoding frame encoding the gene by nucleotide substitution, insertion, deletion, and the like. Therefore, depending on the amino acid substitution, there is no change in the function of the protein, resistance, or positive, harmful, damaging, or disease may be caused.

The composition may be an antibody or an antigen-binding fragment, and may be singular or plural.

“Specifically detecting a polypeptide” means specifically identifying the polypeptide in a sample of an individual.

"Antibody" is a term known in the art and refers to a specific protein molecule directed against an antigenic site. Antibodies can be in the form of whole antibodies as well as functional fragments of antibody molecules. The whole antibody has a structure having two full-length light chains and two full-length heavy chains, and each light chain is connected by a heavy chain and a disulfide bond. A functional fragment of an antibody molecule means a fragment that retains antigen-binding function.

An "antigen-binding fragment" is an entire immunoglobulin structure or a fragment thereof for the entire antibody structure, and refers to a portion of a polypeptide that includes a portion to which an antigen can bind. For example, it may include scFv fragment, (scFv)2 fragment, Fab fragment, Fab' fragment, F(ab')2 fragment, and the like. The antigen-binding fragment can be obtained using a proteolytic enzyme, for example, when the whole antibody is restrictedly digested with papain, Fab can be obtained, and when digested with pepsin, an F(ab')2 fragment can be obtained, and the gene is obtained. It can be produced through recombinant technology.

The composition may be an antigen-antibody binding to a polypeptide containing an amino acid sequence to be detected. The polypeptide is about 1×10 ⁷ M ^-1 , about 1×10 ⁸ M ^-1 , about 1×10 ⁹ M ^-1 , about 1×10 ¹⁰ M ^-1 , or about 1×10 ¹¹ M ^-1 may have binding affinity with an affinity constant (K _A ).

The composition includes immunocytochemistry and immunohistochemistry, radioimmunoassays, Enzyme Linked Immunoabsorbent assay (ELISA), immunoblotting, Farr assay, immunoprecipitation, latex aggregation, Red blood cell aggregation, non-turbidity, immunodiffusion, counter-current electrophoresis, single-radical immunodiffusion, immunochromatography, protein chips, and immunofluorescence can be used. That is, it can be used in a method capable of measuring the binding of an antigen to an antibody.

 Amino acid G is glycine, amino acid A is alanine, amino acid V is valine, amino acid L is leucine, amino acid I is isoleucine, amino acid M is methionine, amino acid F is phenylalanine, amino acid W is tryptophan, amino acid P is proline, amino acid S is serine, Amino acid T is threonine, amino acid C is cysteine, amino acid Y is tyrosine, amino acid N is asparagine, amino acid Q is glutamine, amino acid D is aspartate, amino acid E is glutamate, amino acid K is lysine, amino acid R is arginine, amino acid H Means histidine.

The composition may be a solid composition or a liquid composition. The liquid composition may include the polypeptide in a suitable liquid medium. The liquid medium may be any used in the art as a liquid medium of the polypeptide. In addition, the liquid medium may be appropriately selected according to the purpose of using the composition. For example, the liquid medium can be water or a buffer.

Another aspect provides a kit for diagnosing the risk of developing Sharko-Marie-Tooth comprising the composition. The polypeptide, composition, individual and Sharko-Marie-Tus are as described above. The kit may be a kit for diagnosing the risk of developing Sharko-Marie-Tus in an individual through the presence or absence of the polypeptide. For example, the kit may be a protein chip kit. The protein chip kit may include a substrate, a suitable buffer buffer, a secondary antibody labeled with a chromogenic enzyme or a fluorescent substance, a chromogenic substrate, and the like to detect or measure the binding of the antigen to the antibody. As the base material, a nitrocellulose membrane, a 96-well plate synthesized from polyvinyl resin, a 96-well plate synthesized from polystyrene resin, a slide glass made of glass, etc. may be used, and peroxidase (peroxidase) ), alkaline phosphatase, etc. can be used. In addition, FITC, RITC, etc. may be used as the fluorescent material, and ABTS (2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid)), OPD (o-phenyl) as a chromogenic substrate Rendiamine), TMB (tetramethyl benzidine), and the like.

Another aspect provides a method of determining a nucleotide at a variant position comprising determining a nucleotide at the variant position of the polynucleotide in an isolated biological sample.

The method includes providing an isolated biological sample. Methods of isolating nucleic acids from individuals are known in the art. For example, it can be isolated by directly separating DNA from tissue, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, or urine, or by amplifying specific regions by nucleic acid amplification methods such as PCR. The isolated nucleic acid sample includes not only purely isolated nucleic acid, but also crude isolated nucleic acid, for example, cell debris containing nucleic acid. The nucleic acid amplification methods include PCR, ligase chain reaction (LCR), transcription amplification, self-maintaining sequence replication and nucleic acid based sequence amplification (NASBA). The isolated nucleic acid may be DNA or RNA. The DNA can be genomic DNA, cDNA or recombinant DNA. The RNA may be mRNA.

The method also includes the step of determining the nucleotide at the mutation site of the polynucleotide described above. It is known to determine the nucleotide of a mutation site. For example, the nucleotide of a mutation site can be directly determined by a nucleotide sequencing method of a known nucleic acid. Nucleotide sequencing methods may include Sanger (or dideoxy) sequencing methods or Maksam-Gilbert (chemical cleavage) methods. In addition, a nucleotide at a variable position can be determined by hybridizing a probe containing the sequence at the variable position with a target polynucleotide and analyzing the hybridization result. The degree of hybridization can be confirmed, for example, by labeling a target nucleic acid with a detectable label and detecting the hybridized target nucleic acid, or by an electrical method or the like. In addition, a single base primer extension (SBE) method can be used.

In the method, the step of determining the polynucleotide of the mutation site may include hybridizing the nucleic acid sample to a microarray in which the polynucleotide is immobilized; And detecting the hybridization result.

In addition, the method may include determining that the subject belongs to a risk group having a high probability of developing Sharko-Marie-Tooth if a risk allele exists as a result of determining the nucleotide of the mutation site.

The term "risk allele" refers to the reference allele as the minor allele a, where a in the disease group is greater than the frequency of a in the normal group, or a if the a group in the disease group is the majority allele Can be considered as a risk confrontation. The more risk an allele has, the more likely it is that Sharko-Marie-Tus will develop it. For example, when the minor allele a is present at the variant position of SEQ ID NO: 1, the subject can be determined to belong to a risk group with a high probability of developing Sharko-Marie-Tus. For example, the nucleotide at the determined mutation position is from the 5'end of SEQ ID NO: 1, the 47th nucleotide is G, the 112th nucleotide is T, the 139th nucleotide is A, the 157th nucleotide is C, the 414th nucleotide is G, and If the 328th nucleotide is a nucleotide selected from the group consisting of A, it can be determined that the subject belongs to a risk group with a high probability of developing Sharko-Marie-Tooth.

In the above method, the subject means a subject for predicting the risk of developing Sharko-Marie-Tus. The subject can include a vertebrate, a mammal, or a human ( Homo sapiens ). For example, the human may be Korean.

In addition, the method may be for providing information for predicting the risk of developing an individual's Sharko-Marie-Tus. Providing information for predicting the risk of developing Sharko-Marie-Tus of the subject may be for predicting or diagnosing the relative risk of developing Sharko-Marie-Tus. For example, the risk may be for predicting or diagnosing whether the probability of developing Sharko-Marie-Tus is increased compared to a group having a reference genomic sequence or a normal group. If the number of mutations in the SNP position is 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, or 6, it may be determined that the risk of developing Sharko-Marie-Tus is high, and may be used for prediction or diagnosis. Accuracy and reliability can be increased.

Another aspect provides a method of determining an amino acid at a variant position comprising determining the amino acid at the variant position of the amino acid in an isolated biological sample.

The method includes providing an isolated biological sample. Methods of isolating a sample from an individual are known in the art. For example, tissues, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, or samples such as urine may be included, but is not limited thereto.

The method also includes determining the amino acid at the amino acid substitution position of the polypeptide described above. It is known to determine the amino acid at the amino acid position. For example, a peptide sequencer or protein sequencer can be used to directly determine the amino acid at the amino acid substitution site. In addition, the amino acid sequence can be determined using a protein chip, an enzyme-linked immunoassay, or the like, capable of detecting the amino acid at the amino acid substitution position while measuring the binding of the antigen to the antibody.

In addition, the method may include determining that the subject belongs to a risk group having a high probability of developing Sharko-Marie-Tooth if an amino acid substitution exists as a result of determining the amino acid at the amino acid mutation position.

In the above method, the amino acid at the determined amino acid mutation position is the 16th amino acid R from the N-terminal of SEQ ID NO: 2, the 38th amino acid is L, the 47th amino acid is K, the 53th amino acid is R, the 138th amino acid is R, 110 If the second amino acid is S, or a combination thereof, the subject can be determined to belong to a risk group with a high probability of developing Sharko-Marie-Tooth.

In the above method, the subject refers to a subject for predicting the risk of developing Sharko-Marie-Tus. The subject may include vertebrates, mammals, or humans ( Homo sapiens ). For example, the human may be Korean.

Providing information for predicting the risk of developing Sharko-Marie-Tus of the subject may be for predicting or diagnosing the relative risk of developing Sharko-Marie-Tus. For example, the risk may be for predicting or diagnosing whether the probability of developing Sharko-Marie-Tooth is increased compared to a group having a reference genomic sequence or a normal group. When the number of mutations in the mutation position is 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, or 6, it may be determined that the risk of developing Sharko-Marie-Tus is high, and prediction or diagnosis The accuracy and reliability of can be increased.

The polypeptide comprising the amino acid sequence of SEQ ID NO: 2 may be a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 1, a protein registered with NCBI Accession number NP_000157.1, or a human GJB1 protein.

The GJB1 protein is a transmembrane protein located across EC (extracellular, extracellular), TM (transmembrane), and IC (intracellular, intracellular). The p.H16R mutation may be located in the IC domain of the GJB1 protein. The p.V38L mutation may be located in the TM domain of the GJB1 protein. The p.E47K mutation may be located in the EC domain of the GJB1 protein. The p.C53R mutation may be located in the EC domain of the GJB1 protein. The p.S138R mutation may be located in the TM domain of the GJB1 protein. The p.G110S mutation may be located in the IC domain of the GJB1 protein.

In addition, the method may include determining that the subject belongs to a risk group having a high probability of developing Sharko-Marie-Tooth if an amino acid substitution exists in the EC domain as a result of determining the amino acid at the amino acid mutation position. .

According to the method, when the amino acid at the amino acid mutation position is determined, when the amino acid substitution is present in the EC domain, compared to the case where the amino acid substitution is present in the IC domain or TM domain, the individual is severely affected by the development of Sharco-Marie-Tooth. It may include the step of determining to exhibit the symptoms of.

The compositions and kits can be used to quickly and efficiently diagnose the risk of developing Sharko-Marie-Tus. In addition, according to the method for providing information for predicting the risk of developing Sharko-Marie-Tus, it is possible to efficiently check whether the individual has a risk of developing Sharko-Marie-Tus.

FIG. 1 shows the structure, domain and distribution of 43 mutations of the GJB1 protein, identified in families including Korean CMTX1 patients. The structure and domain of the GJB1 protein is Kleopa et al.(Kleopa et al. How do mutations in GJB1 cause X-linked Charcot-Marie-20 Tooth disease? Brain Res 2012;1487:198-205).
2 is a result of confirming whether the sequence is conserved by arranging the amino acid sequence of the position of 17 mutations and the surrounding region including the same in some vertebrate species. The vertebrate species is human (Homo sapiens), Rat (Mus musculus), rat (Rattus norvegicus), A wolf (Canis lupus familiaris), cow (Bos taurus), Chicken (Gallus gallus) And African clawed frogs (Xenopus laevis)to be. The amino acid sequence was obtained from NCBI, and the accession number of each sequence wasHomo sapiens: NP_000157.1, Mus musculus: NP_001289425.1,Rattus norvegicus: NP_058947.1,Canis lupus familiaris: XP_538073.1,Bos taurus: NP_776494.1, Gallus gallus: NP_989702.1, andXenopus laevis: Corresponds to NP_001095219.1. Conservation analysis of the protein sequence was performed using MEGA6, version 6.0 (http://www.megasoftware.net/).
Figure 3 shows the association between the severity of the disease and the duration of the disease, Figure 3A shows the relationship between the disease duration and FDS, Figure 3B is a graph showing the relationship between the disease duration and CMTNS.
4 is a graph showing the distribution of the onset age of Korean CMTX1 patients.

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

Example  1: Variation position in patient group and normal group Allele  Through analysis Sharko -Associated with Marie-Tooth Marker  Selection

In this example, in the genomes of Charco-Marie-Tus patients and normal persons, alleles of mutation sites were analyzed, and from the results, mutation markers associated with Charco-Marie-Tus were selected.

(1) Selection of test subjects

In the Korean 655 family, 1092 and 952 family members with CMT were selected as subjects of this study. Among them, a total of 128 patients, 67 males and 61 females were selected from 63 families with a family history of Sharko-Marie-Tus. In addition, 78 patients, 35 males and 43 females, and 278 healthy males, 278 males and 222 females, who were not affected by the disease, were included in the study. The protocol of this study was approved by the clinical research deliberation committee of Ewha Womans University Mokdong Hospital and Sungkyunkwan University Samsung Seoul Hospital.

(2) DNA purification and 17p12  Through prescreening of duplicates GJB1  Genetic analysis

Genomic DNA was purified from the peripheral blood of the individual of (1) using QIAamp blood DNA purification kit (Qiagen, Germany). Whether the 17p12 (PMP22) overlap/deletion in the sample is hexaplexx microsatellite polymerase chain reaction (PCR) and real-time polymerase chaining Screening was performed using a reaction (real-time PCR). Peripheral neuropathy relevant gene by sequencing GJB1 mutations by direct sequencing, whole exome sequencing, and a panel of genes containing 73 genes. -targeted sequencing) method. It was confirmed that the 128 persons had a mutation in the GJB1 gene. If the mutation of the GJB1 gene was recently reported or reported in the Inherited Peripheral neuropathy Mutation Database (http://www.molgen.ua.ac.be/CMTMutations/Mutations/), pathogenicity Was judged. In addition, it was determined that there is a possibility of etiology when a new mutation is not detected in 500 healthy control subjects, and is co-segregated from member individuals affected by the disease in the household.

(3) Clinical evaluation method

Clinical data include gender, age at onset, duration of illness, and presence of motor loss, sensory loss, areflexia, foot deformities, and muscle wasting. . Physical disabilities were measured by a functional disability scale (FDS) and a CMT neuropathy score (CMTNS). Sensorineural hearing loss or hearing loss was measured by brainstem auditory evoked potentials (BAEP).

(4) Electrophysiology ( electrophysiological ) Analysis method

Electrophysiological analysis was performed on the Sierra Wave EMG system (Cadwell Lab, Kennewick, WA, USA), Medelec Synergy (Oxford Instruments Medical, Hawthorne, NY, USA), and Toungies 2- It was carried out using a channel neuroscreen system (Toennies two-channel NeuroScreen system) (Jaeger-Toennies, Hochberg, Germany). Limb movement and sensory NCS (nerve conduction study) was recorded using conventional methods (Oh SJ. Clinical Electromyography: Nerve Conduction Studies. Lippincott Williams & Wilkins, 2003).

NCS was performed with a surface electrode. CMAP for each of the abductor pollicis brevis and adductor digiti quinti, as measured by stimulation at the elbow or wrist, MNCV (motor nerve conduction velocity) of the median and ulnar nerves (Compound muscle action potential) was recorded. In the same way, MNCV and CMAP of the fibula nerve and tibia nerve were measured. SNCV (sensory nerve conduction velocity) and SNAP (sensory nerve action potential) were obtained for the finger-wrist region from the median nerve and ulnar nerve. Needle EMG (needle electromyography) was performed on the bilateral proximal and distal lower extremity muscles.

(5) Statistical analysis

Chi-square, Kruskal-Wallis, and Fisher's exact test were used to compare discrete variables. In addition, the independent sample T-test and Mann-Whitney U test were used to compare the two samples against the average value of the continuous variables. Pearson's correlation coefficient was calculated based on FDS values and clinical variables. When the p-value was less than 0.05, the difference was judged to be statistically significant. Statistical analysis was performed using a statistical package from Social Sciences (SPSS) version 18.0 (IBM, Armonk, NY, USA).

(6) Korean CMTX1  Analysis of patient's mutation profile

Samples of 128 patients with mutations in the GJB1 gene in 63 families with a family history of Sharko-Marie-Tus were analyzed. The frequency of CMTX1 is about 9.6%, and this number is higher than the frequency suggested in the previous report, about 5.3%, and it is about 14.8% when calculated based on genetically identified CMT cases.

A total of 43 etiological variations were identified in 63 families. The pathogenic mutations are 39 missense mutations and 4 termination/lattice shift mutations. The variation is shown in Table 1, Table 2, Table 3 and FIG. 1. Variations specifically identified for Korean patients are underlined. The p.V95M mutation was most frequently seen in families including Korean CMTX1 patients. Standard measurement errors are indicated as ±.

Seventeen variations that were not reported in non-Korean races were located in well-conserved regions of the amino acid sequence. 2 is a result of analyzing the degree of sequence preservation of a region containing 17 mutations.

transition Domain ^b GERP ^c Double Rico (In- silico) predicted ^d ancestry
ID Additional symptoms ^e Severity ^f The onset age Examination age Nucleotide ^a amino acid S P2 Mu FDS CMTNS c.3G>T p.M1I IC 4.15 0.00 0.998 -0.422 FC582 HAF, stroke 0 5 10 15 c.7T>C p.W3R IC 4.15 0.00 1.000 -0.796 FC323 HAF 2 9 15 24 c.13G>A p.G5S IC 4.15 0.00 0.044 -0.588 FC143 HAF 2 - 15 19 c.20A>G p.Y7C IC 4.15 0.00 1.000 0.013 FC718 HAF, ataxia, tremor 2 - 15 35 c.22A>C p.T8P IC 2.98 0.25 0.018 -0.124 FC507 HAF 3 12 22 26 FC602 HAF 2 8 47 52 c.43C>T p.R15W IC 4.36 0.00 1.000 -0.873 FC751 - - - - - c.44G>A p.R15Q IC 4.36 0.06 0.995 -1.000 FC583 HAF, ataxia, tremor One 17 27 36 c.47A>G p.H16R IC 4.36 1.00 0.999 -0.177 FC584 HAF 2 8 10 51 c.62G>A p.G21D TM 4.36 0.00 1.000 -0.195 FC068 HAF, ataxia One 15 47 50 c.77C>T
p.S26L
TM
4.36
0.00
0.999
-0.273
FC120 HAF One 8 37 42 FC381 HAF 3 14 12 26 c. 77delC p.S26fs TM 4.36 . . . FC267 HAF, tremor 2 8 14 20 c.109G>T p.V37L TM 4.26 0.00 1.000 -0.594 FC258 HAF, ataxia, tremor One 10 7 21 c.112G>T p.V38L TM 4.26 0.06 0.443 0.320 FC491 HAF 2 10 15 19 c.137A>G p.D46G EC 4.56 0.01 1.000 -0.811 FC539 HAF, ataxia, tremor 3 26 15 33 c.139G>A p.E47K EC 4.56 0.00 1.000 -0.811 FC635 HAF 3 12 48 54

transition Domain ^b GERP ^c Double Rico (In- silico) predicted ^d ancestry
ID Additional symptoms ^e Severity ^f The onset age Examination age Nucleotide ^a amino acid S P2 Mu FDS CMTNS c.157T>C p.C53R EC 4.56 0.00 1.000 -0.442 FC476 HAF, tremor 3 14 15 18 c.257C>T p.T86I TM 4.67 0.00 1.000 -0.025 FC382 Mild 2 11 8 19 c.271G>A p.V91M TM 4.67 0.03 1.000 -0.448 FC501 HAF, SNHL 3 12 9 68 c.283G>A p.V95M IC 4.67 0.00 1.000 -0.219 FC134 HAF, ataxia, tremor 3 13 21 25 FC565 HAF, ataxia 2 9 48 56 FC580 HAF 4 22 9 43 FC622 HAF, ataxia One 15 8 23 FC687 HAF, ataxia, tremor One 4 8 22 FC698 Ataxia 2 11 40 50 c.286G>C p.A96P IC 4.67 0.00 1.000 -0.694 FC725 HAF, ataxia, tremor 2 - 11 22 c.328G>A p.G110S IC 4.67 - 0.025 -1.000 FC788 - - - - - c.394T>C p.W132R TM 4.67 0.00 1.000 -0.905 FC069 HAF, tremor 3 12 15 34 FC277 HAF, ataxia, tremor One 12 14 21 FC799 - - - - - c.407T>C p. V136A TM 4.67 0.00 0.007 -1.000 FC002 3 17 One 13 c.414C>G p. S138R TM -2.68 0.01 1.000 0.427 FC370 HAF One 5 39 40 c.415G>A p.V139M TM 3.81 - 0.994 -0.777 FC781 - - - - - c.424C>T p.R142W TM 4.67 0.00 1.000 -0.267 FC674 HAF 2 9 20 25 c.425G>A p.R142Q TM 4.67 0.00 1.000 -0.677 FC194 HAF, ataxia, tremor 3 9 22 24 c.451T>C
p.Y151H
EC
4.67
0.00
1.000
-0.910
FC289 HAF, ataxia, tremor 6 21 35 43 FC743 HAF 3 4 25 47 c. 454delG p.V152fs EC 4.67 . . . FC312 HAF, ataxia, tremor 6 19 12 52 c.458T>G p.F153C EC 4.67 0.00 1.000 -1.000 FC115 HAF 0 4 35 47 FC663 Mild 2 10 21 26 c.476G>A p.G159D EC 4.81 0.00 0.775 -0.004 FC251 HAF 3 19 27 65

transition Domain ^b GERP ^c Double Rico (In- silico) predicted ^d ancestry
ID Additional symptoms ^e Severity ^f The onset age Examination age Nucleotide ^a amino acid S P2 Mu FDS CMTNS c.490C>T p.R164W EC 3.91 0.00 0.993 -0.818 FC298 HAF 3 15 22 35 FC412 Mild 3 15 15 51 FC705 HAF, ataxia 2 - 55 60 FC714 HAF, ataxia, tremor 4 15 30 55 c.491G>A p.R164Q EC 4.81 0.00 1.000 -0.953 FC030 HAF, tremor, Contr 3 13 12 19 FC163 HAF, ataxia, VCP, Hoarse, Myot 3 18 15 22 FC254 Tremor, pain One - 15 65 FC367 HAF 2 9 15 21 FC722 HAF, ataxia 3 20 8 36 c.502T>C p.C168R EC 4.81 0.00 1.000 -0.687 FC022 HAF, tremor, SNHL 4 21 26 36 c.536_ 537insCTGA p.C179X EC 4.99 . . . FC359 HAF, ataxia, tremor 4 18 20 26 c.538T>C p.F180L EC 4.99 0.00 1.000 -1.000 FC517 HAF, ataxia, tremor, Contr One 13 11 20 FC662 HAF 3 12 9 40 c.548G>A p.R183H EC 4.99 0.00 1.000 -0.442 FC481 HAF, ataxia, tremor 3 17 19 24 FC653 HAF, tremor, SNHL One 11 20 35 c.551C>T p.P184L EC 4.99 0.00 1.000 0.439 FC147 Mild 2 - 12 28 c.556G>A p.E186K EC 4.13 0.00 1.000 -0.835 FC571 HAF 0 2 5 7 c.590C>T p.A197V T M 4.99 0.00 0.012 0.052 FC074 HAF 4 14 26 36 c.622G>A p.E208K IC 4.76 0.00 1.000 -0.876 FC224 HAF 3 15 25 59 c.641-642TC>AT p.I214N IC 4.90 0.00 0.007 -1.000 FC095 HAF, ataxia, tremor 6 19 21 25 c.658C>T p.R220X IC 1.96 . . . FC271 HAF 2 12 45 71

The term is defined as follows.

IC: intracellular domain

EC: extracellular domain

TM: transmembrane domain

GERP: the genomic evolutionary rate profiling figure

S: SIFT, <0.05, affects protein function (http://sift.jcvi.org)

P2: polyphen 2, confidence level 0>1 (http://genetics.bwh.harvard.edu/pph2/)

Mu: MUpro,-decreases protein stabilization, + increases protein stabilization

HAF: high arched foot

Contr: Construction

SNHL: sensorineural hearing loss

VCP: vocal cord palsy

Hoarse: speech disorder (hoarseness)

myt dys: myotonic dystrophy

FDS: functional disability scale

CMTNS: CMT neuropathy score

(7) Korean CMTX1  Analysis of clinical characteristics of patient mutation

Of the 61 female patients, 22 were asymptomatic. Of the 128 patients, 89 patients with symptoms and electrophysiological records were selected. High-arched feet were observed in 87% of patients, ataxia in 41% of patients, and tremor in 35% of patients. Sensorineural hearing loss was observed in 4 patients with p.C168R, p.V91M, p.D46G, and p.R183H mutations. However, optic atrophy, scoliosis, spasticity and painful sensory paresthesia were not observed. The clinical characteristics are shown in Table 4 below.

The degree of disability varies from patient to patient, but the average values of FDS and CMTNS were 2.3 and 12.2, respectively, indicating moderate severity. None of the patients depended on wheelchairs or used crutches. In evaluating the disorder, FDS and CMTNS were highly correlated ( r =0.723, p <0.001), and disease severity was related to the duration of disease in all male patients. CMTNS levels were associated with symptomatic female patients. 3A is a graph showing the relationship between the disease period and FDS, and FIG. 3B is a graph showing the relationship between the disease period and CMTNS.

The phenotype of CMTX1 patients showed significant differences according to gender. 36% of asymptomatic female patients and symptomatic female patients showed a mild degree of phenotype compared to each of asymptomatic male patients and symptomatic male patients.

The age of onset was significantly different by gender. The age of onset in male patients developed at a younger age compared to the distribution of age onset in female patients. 4 is a graph showing the distribution of the onset age of Korean CMTX1 patients. Looking at the age at which symptoms developed, 29.7% of male patients (19 out of 64) were before the age of 10, and 9.4% of male patients (6 out of 64) were after the age of 30.

Clinical features
(Clinical manifestation) gun man Woman p-number Total (number of patients n) 128 67 (52%) 61 (48%) Symptoms (Symptomatic) 106 (83%) 67 (100%) 39 (64%) <0.001 No symptoms (Asymptomatic) 22 (17%) 0 (0%) 22 (36%) Data analysis 89 58 31 Test age (yr) 37.4±17.0 34.7±16.8 42.5±16.5 0.040 Onset age (yr)
Disease period (yr) 23.3±14.6
14.2±13.4 18.4±11.1
16.3±14.3 32.1±16.1
10.3±10.7 <0.001
0.047 FDS 2.3±1.5 2.6±1.3 1.7±1.6 0.004 CMTNS 12.2±5.8 13.2±5.4 10.2±5.9 0.019 Foot deformity (n, %) 76 (87%) 54 (96%) 22 (71%) 0.001 Ataxia (n, %) 36 (41%) 29 (52%) 7 (23%) 0.008 Tremor (n, %) 30 (35%) 24 (43%) 6 (19%) 0.027 Scoliosis (n) 0 0 0 Contract (n, %) 2 (2%) 2 (4%) 0 (0%) 0.536 Optic atrophy (n) 0 0 0 Vocal cord palsy
(n, %) 4 (5%) 3 (5%) 1 (3%) 0.454 Vocal cord palsy
(n, %) 1 (1%) 1 (2%) 0 (0%) 0.649 Spasticity (n) 0 0 0 Painful luck
(Painful sensory paresthesia) (n) 0 0 0 Wheelchair-bound (n) 0 0 0

The term is defined as follows.

FDS: Functional Disability Scale

CMTNS: CMT neuropathy score (Charcot.Marie.Tooth neuropathy score)

SNHL: sensorineural hearing loss

The electrophysiological characteristics of Korean CMTX1 patients are shown in Table 5 below. Male patients showed a more severe phenotype in electrophysiology data. FDS and CMTNS showed statistically significant differences between male and female patients. Motor NCS (CMAP and NCV) showed significant differences between male and female patients, except for CMAP for tibial nerves. In addition, the incidence of foot anomalies, ataxia and hydrocephalus showed significant differences between male and female patients (see Tables 4 and 5). The mean FDS was 2.6 in male patients and 1.7 in female patients with symptoms ( p =0.004). The mean value of CMTNS was 13.2 in male patients and 10.2 in female patients with symptoms ( p = 0.019).

According to the results of electrophysiological analysis, the median nerve NCV and CMAP mean amplitudes were 31.33 m/s and 4.28 mV for male patients, respectively, while 40.75 m/s and 6.66 mV for symptomatic female patients, respectively. There was a significant difference. This difference was also observed for motor NCS of the ulna and fibula nerves.

53.6% of male patients had moderate NCV (30-40 m/s), 20.7% of male patients had mild (>40 m/s), and 15.6% of male patients had severe slowing (<30 m /s). 48.4% of female patients with symptoms slowed in mildness (>40 m/s), 38.7% of female patients with symptoms had moderate NCV (30-40 m/s), and 12.9% of female patients with symptoms were severe Slowing (<30 m/s) was shown. Each data is expressed as ± standard error (SEM).

Electrophysiological features Total (n=89) Male (n=58) Female (n=31) p-number Median motor nerve ^a MNCV (m/s) 37.26±7.80 31.33±12.33 40.75±9.09 <0.001 CMAP (mV) 5.13±4.72 4.28±4.12 6.66±5.39 0.024 No reaction (n, %) 6 (7%) 6 (11%) 0 (0%) 0.059 Ulnar motor nerve MNCV (mV) 40.47±8.05 37.16±4.83 46.46±9.23 <0.001 CMAP (m/s) 8.60±3.89 7.90±3.64 9.86±4.06 0.023 No reaction (n, %) 0 (0%) 0 (0%) 0 (0%) Peroneal nerve MNCV (m/s) 15.25±18.84 9.16±14.93 26.26±20.36 <0.001 CMAP (mV) 1.21±2.18 0.54±1.26 2.44±2.89 0.001 No reaction (n, %) 51 (58.6%) 40 (71.4%) 11 (35.5%) 0.002 Tibial nerve MNCV (m/s) 25.93±14.47 23.03±14.83 31.18±12.37 0.008 CMAP (mV) 3.65±6.22 2.93±5.91 4.95±6.65 0.166 No reaction (n, %) 17 (19.5%) 14 (25%) 3 (9.7%) 0.098 Median sensory nerve
SNCV (m/s) 19.45±15.18 18.68±15.29 20.98±15.11 0.500 SNAP (μV) 4.65±5.44 4.08±5.30 5.69±5.63 0.188 No reaction (n, %) 32 (37%) 22 (39%) 10 (32%) 0.515 Ulnar sensory nerve SNCV (m/s) 19.97±14.49 19.23±14.66 21.32±14.31 0.523 SNAP (μV) 4.18±4.83 3.72±4.86 5.03±4.72 0.228 No reaction (n, %) 29 (33%) 20 (36%) 9 (29%) 0.527 Sural nerve SNCV (m/s) 13.02±13.86 11.39±13.03 15.96±15.01 0.160 SNAP (μV) 3.38±5.254 2.59±4.23 4.81±6.56 0.096 No reaction (n, %) 45 (51.7%) 31 (55.4%) 14 (45.2%) 0.362

The term is defined as follows.

MNCV: motor nerve conduction velocity

CMAP: compound muscle action potential

SNCV: sensory nerve conduction velocity

SNAP: sensory nerve action potential

The relationship between CMAP of the median, ulnar and tibial nerves, motor NCV of the median, ulnar, fibula and tibial nerves, and SNAP of the ulnar nerve were confirmed. For male patients, the duration of the disease is CMAP of the median, ulnar and fibula nerves; It was inversely correlated with motor NCV of the ulnar nerve and not related to the sensory nerve. In female patients with symptoms, the duration of the disease is CMAP of the median and ulnar nerves; Median and tibial nerve motors were associated with NCV. CMTNS correlated with all amplitudes and velocities in the upper and lower extremities, except for the tibial nerve in male patients. Table 6 shows the relationship between the parameters related to the disease.

parameter Disease period FDS CMTNS Pearson correlation
(Pearson correlation) Note
(2-tailed) Pearson
correlation Note
(2-tailed) Pearson
correlation Note
(2-tailed) Disease period One - 0.290 0.027 0.266 0.047 FDS 0.290 0.027 One - 0.723 0.000 CMTNS 0.266 0.047 0.723 0.000 One - Polite CMAP -0.273 0.038 -0.485 0.000 -0.511 0.000 Courteous MNCV -0.185 0.163 -0.222 0.094 -0.331 0.013 Ulna CMAP -0.372 0.004 -0.480 0.000 -0.484 0.000 Ulnar MNCV -0.349 0.007 -0.111 0.408 -0.338 0.011 Polite SNAP -0.143 0.284 -0.361 0.005 -0.504 0.000 Polite SNCV -0.063 0.638 -0.275 0.037 -0.444 0.001 Ulna SNAP -0.161 0.228 -0.288 0.028 -0.440 0.001 Ulnar SNCV -0.161 0.228 -0.206 0.120 -0.426 0.001 Fibula MNCV -0.216 0.104 -0.375 0.004 -0.438 0.001 Fibula CMAP -0.275 0.037 -0.345 0.008 -0.344 0.009 Tibia MNCV -0.142 0.287 -0.116 0.386 -0.211 0.119 Tibial CMAP -0.250 0.059 -0.225 0.089 -0.160 0.238 Raincoat MNCV -0.077 0.564 -0.334 0.010 -0.402 0.002 Zebu SNAP -0.107 0.422 -0.246 0.062 -0.365 0.006

(8) Analysis of patients with no symptoms

Patients with no symptoms were defined as follows. No symptoms are 1) the patient's electrophysiological data is in the normal category and 2) no clinical symptoms such as high arched feet.

Twenty-two patients with CJB1 mutations (M1I, W3R, R15Q, D46G, T86I, V95M, W132R, R142Q, R142W, F153C, R164Q, R164W, F180L, A197V, and I214N) showed no symptoms and were all female patients. Fourteen patients were confirmed by neurological examination and electrophysiological analysis. Eight patients were confirmed by neurological examination and clinical features. In male patients with the same mutations as female patients with no symptoms, the onset age did not show a significant difference compared to male patients with other mutations (21.8 vs 25.3, p=0.27). In addition, the two male patients showed no significant difference in severity.

(9) Korean CMTX1  Analysis of patient genotype-phenotype association

The genotype-phenotype association of CMTX1 induced by GJB1 mutation was analyzed. Since CMTX1 is associated with the X chromosome, it can be seen that there is a difference in genetic composition between male and female patients with this disease. All female patients were heterozygous for mutation, but all male patients were hemizygous. By lyonisation, women have randomly inactivated mutations and wild-type X chromosomes, so it is expected to show differences in the severity of sex between men and women in clinical characteristics.

Electrophysiological data of a total of 58 male patients was collected and classified into 3 domain groups according to the mutation location. The characteristics of phenotype according to domain analysis in male CMTX1 patients are shown in Table 7 below. Patients with mutations in the EC domain showed severe phenotypes in FDS, CMTNS, motor NCV, and CMAP. FDS and CMAP of the median and ulnar nerves showed significant differences in patients with mutations in the EC domain and IC domain. In addition, CMTNS and tibial nerve MNCV showed significant differences in patients with mutations in TM domain and EC domain.

The same analysis as above was performed on the female patient. As a result, results similar to those in male patients were confirmed. CMAP of the ulnar nerve, sensory NCV, and SNAP of the ulnar sensory nerve and the median sensory nerve were significantly decreased in patients with mutations in the EC domain compared to patients with mutations in the IC domain.

Table 8 shows the results of analyzing the domains of female CMTX1 patients. Variations in the EC domain showed a severe phenotype compared to variations in the other two domains. When there was variation in the EC domain, 26.1% of female patients with variation in the EC domain were asymptomatic. On the other hand, when the TM domain and the IC domain had a variation, 38.1% and 47.1% of the female patients having a variation in the TM domain and the IC domain, respectively, had no symptoms. The ratio of symptomatic female CMTX1 patients to symptomatic female CMTX1 patients in each domain is shown in Table 9 below.

Characteristic IC domain
(n=20) TM domain
(n=14) EC domain
(n=24) p -number ^a p -number ^b p -number ^c Examination age 37.6±18.7 27.0±14.5 36.5±15.7 0.065 0.922 0.071 The onset age 20.4±14.2 14.0±6.2 19.2±10.1 0.094 0.750 0.289 Disease period 17.3±15.8 13.0±15.0 17.3±13.0 0.245 0.883 0.377 FDS 2.3±1.4 2.4±1.0 3.0±1.4 0.079 0.027 0.572 CMTNS 11.9±5.4 12.1±3.8 14.9±5.9 0.030 0.062 0.862 Foot deformity
(Foot deformity) 18 (95%) 13 (100%) 23 (96%) 1.000 1.000 1.000 Ataxia
(Ataxia) 11 (58%) 6 (46%) 12 (50%) 0.823 0.606 0.513 Tremor 8 (42%) 5 (39%) 11 (46%) 0.666 0.807 0.837 Scoliosis
(Scoliosis) 0 0 0 - - - build
(Contractu re) 0 0 2 (8%) 0.532 0.495 - Optic nerve atrophy
(Optic atrophy) 0 0 0 - - - SNHL 0 1 (8%) 2 (8%) 1.000 0.495 0.406 Vocal cord paralysis
(Vocal cord palsy) 0 0 1 (4%) 1.000 1.000 - Median motor nerve ^d MNCV (m/s) 35.41±4.48 34.69±11.66 26.28±15.18 0.052 0.053 0.328 CMAP (mV) 6.34±4.04 4.64±3.83 2.48±3.62 0.062 <0.001 0.219 no response 0 1 (8%) 5 (21%) 0.394 0.056 0.406 Ulnar motor nerve MNCV (mV) 38.13±4.26 37.58±4.46 36.16±5.40 0.504 0.167 0.502 CMAP (m/s) 9.76±2.86 8.00±3.26 6.37±3.79 0.127 0.002 0.156 no response 0 0 0 - - - Peroneal nerve MNCV (m/s) 7.09±14.33 11.85±15.72 9.33±15.34 0.640 0.627 0.382 CMAP (mV) 0.33±0.72 0.76±1.37 0.58±1.53 0.727 0.509 0.311 no response 15 (79%) 8 (62%) 17 (71%) 0.564 0.545 0.427 Tibial nerve MNCV (mV) 24.02±13.47 31.21±10.18 17.80±16.20 0.004 0.177 0.114 CMAP (m/s) 2.47±4.40 3.44±3.80 3.03±7.78 0.859 0.784 0.526 no response 4 (21%) 1 (8%) 9 (38%) 0.065 0.324 0.625 Median sensory nerve SNCV (m/s) 20.61±14.57 24.03±13.85 14.25±15.91 0.091 0.318 0.295 SNAP (μV) 3.79±3.70 5.36±4.57 3.61±6.65 0.052 0.163 0.269 no response 6 (32%) 3 (23%) 13 (54%) 0.091 0.139 0.704 Ulnar sensory nerve SNCV (m/s) 20.01±14.32 21.34±14.92 17.47±15.19 0.326 0.940 0.402 SNAP (μV) 3.41±3.54 3.92±3.98 3.84±6.20 0.452 0.547 0.26 no response 6 (32%) 4 (31%) 10 (42%) 0.724 0.497 1.000 Sural nerve SNCV (m/s) 14.04±13.78 10.62±12.36 9.71±12.99 0.837 0.297 0.470 SNAP (μV) 3.29±5.55 2.57±3.76 2.04±3.27 0.657 0363 0.688 no response 9 (47%) 7 (54%) 15 (63%) 0.609 0.321 0.719

Characteristic IC domain (N=8) TM domain (N=10) EC domain (N=13) p -number ^a p -number ^b p -number ^c Examination age 43.00±22.08 38.60±16.95 45.08±12.56 0.214 0.942 0.534 The onset age 34.38±22.29 32.10±13.35 30.77±14.66 0.950 0.799 0.755 Disease period 8.62±8.14 6.50±8.94 14.31±12.60 0.047 0.292 0.304 FDS 2.13±1.89 1.20±0.92 1.69±1.84 0.796 0.502 0.282 CMTNS 10.75±6.67 8.40±5.91 11.31±5.59 0.277 1.000 0.422 Foot deformity 6 (75%) 7 (70%) 9 (69%) 1.000 1.000 1.000 Ataxia 4 (50%) 2 (20%) 1 (8%) 0.560 0.047 0.321 Tremor 2 (25%) 2 (20%) 2 (15%) 1.000 0.618 1.000 Scoliosis
(Scoliosis) 0 0 0 - - - build
(Contractu re) 0 0 0 - - - Optic nerve atrophy
(Opticatrophy) 0 0 0 - - - SNHL 0 0 1 (8%) 1.000 1.000 Vocal cord paralysis
(Vocal cord palsy) 0 0 0 - - - Median motor nerve ^d MNCV (m/s) 43.05±6.76 39.97±9.32 39.94±10.49 0.877 0.365 0.424 CMAP (mV) 7.19±4.65 7.72±6.94 5.52±4.62 0.515 0.469 0.929 no response 0 0 0 - - - Ulnar motor nerve MNCV (mV) 48.79±7.87 48.35±11.13 43.58±8.28 0.264 0.148 0.563 CMAP (m/s) 11.69±2.51 10.37±4.47 8.35±4.21 0.278 0.046 0.248 no response 0 0 0 - - - Peroneal nerve MNCV (m/s) 31.31±19.67 27.92±20.17 21.88±21.57 0.523 0.314 0.753 CMAP (mV) 2.83±2.34 1.94±2.28 2.58±3.67 0.949 0.628 0.419 no response 2 (25%) 3 (30%) 6 (46%) 0.669 0.400 1.000 Tibial nerve MNCV (mV) 29.93±14.63 33.13±13.29 30.45±10.98 0.456 0.971 0.563 CMAP (m/s) 5.74±6.37 6.99±8.97 2.88±4.27 0.926 0.717 0.689 no response 1 (13%) 1 (10%) 1 (8%) 1.000 1.000 1.000 Median sensory nerve SNCV (m/s) 27.70±11.33 20.11±17.57 17.52±14.85 0.482 0.014 1.000 SNAP (μV) 9.65±6.28 5.71±5.74 3.23±3.82 0.250 0.006 0.164 no response 1 (13%) 4 (40%) 5 (39%) 1.000 0.336 0.314 Ulnar sensory nerve SNCV (m/s) 25.43±10.66 22.89±16.35 17.58±14.72 0.217 0.256 0.655 SNAP (μV) 8.08±5.63 4.77±4.47 3.65±3.58 0.466 0.028 0.195 no response 1 (13%) 3 (30%) 5 (39%) 1.000 0.336 0.588 Sural nerve SNCV (m/s) 25.46±10.42 15.77±16.81 10.26±13.89 0.338 0.013 0.526 SNAP (μV) 10.41±8.92 4.24±4 1.80±3.38 0.273 0.004 0.124 no response 1 (13%) 5 (50%) 8 (62%) 0.580 0.067 0.152

The term is defined as follows.

EC: extracellular

TM: transmembrane

IC: intracellular

FDS: Functional Disability Scale

CMTNS: CMT neuropathy score (Charcot-Marie-Tooth neuropathy score)

SNHL: sensorineural hearing loss

MNCV: motor nerve conduction velocity

CMAP: compound muscle action potential

SNCV: sensory nerve conduction velocity

SNAP: sensory nerve action potential

a: Analysis between TM and EC domain groups

b: Analysis between IC and EC domain groups

c: Analysis between TM and IC domain groups

Data in the absence of response were expressed as 0 m/s (NCV), 0 mV (CMAP), and 0 μV (SNAP).

patient domain gun p -number ^* p -number ^a p -number ^b p -number ^c IC TM EC Symptoms (Symptomatic) 9 13 17 39 0.383 0.393 0.169 0.578 No symptoms (Asymptomatic) 8 8 6 22 ratio 47.1% 38.1% 26.1% 36.1%

*: Analysis between IC, TM and EC domain groups

a: Analysis between TM and EC domain groups

b: Analysis between IC and EC domain groups

c: Analysis between TM and IC domain groups

<110> Samsung Life Public Welfare Foundation KONGJU NATIONAL UNIVERSITY INDUSTRY-UNIVERSITY COOPERATION FOUNDATION <120> Marker for diagnosing Charcot-Marie-Tooth and use thereof <130> PN115854 <160> 2 <170> KopatentIn 2.0 <210> 1 <211> 852 <212> DNA <213> Homo sapiens <400> 1 atgaactgga caggtttgta caccttgctc agtggcgtga accggcattc tactgccatt 60 ggccgagtat ggctctcggt catcttcatc ttcagaatca tggtgctggt ggtggctgca 120 gagagtgtgt ggggtgatga gaaatcttcc ttcatctgca acacactcca gcctggctgc 180 aacagcgttt gctatgacca attcttcccc atctcccatg tgcggctgtg gtccctgcag 240 ctcatcctag tttccacccc agctctcctc gtggccatgc acgtggctca ccagcaacac 300 atagagaaga aaatgctacg gcttgagggc catggggacc ccctacacct ggaggaggtg 360 aagaggcaca aggtccacat ctcagggaca ctgtggtgga cctatgtcat cagcgtggtg 420 ttccggctgt tgtttgaggc cgtcttcatg tatgtctttt atctgctcta ccctggctat 480 gccatggtgc ggctggtcaa gtgcgacgtc tacccctgcc ccaacacagt ggactgcttc 540 gtgtcccgcc ccaccgagaa aaccgtcttc accgtcttca tgctagctgc ctctggcatc 600 tgcatcatcc tcaatgtggc cgaggtggtg tacctcatca tccgggcctg tgcccgccga 660 gcccagcgcc gctccaatcc accttcccgc aagggctcgg gcttcggcca ccgcctctca 720 cctgaataca agcagaatga gatcaacaag ctgctgagtg agcaggatgg ctccctgaaa 780 gacatactgc gccgcagccc tggcaccggg gctgggctgg ctgaaaagag cgaccgctgc 840 tcggcctgct ga 852 <210> 2 <211> 283 <212> PRT <213> Homo sapiens <400> 2 Met Asn Trp Thr Gly Leu Tyr Thr Leu Leu Ser Gly Val Asn Arg His   1 5 10 15 Ser Thr Ala Ile Gly Arg Val Trp Leu Ser Val Ile Phe Ile Phe Arg              20 25 30 Ile Met Val Leu Val Val Ala Ala Glu Ser Val Trp Gly Asp Glu Lys          35 40 45 Ser Ser Phe Ile Cys Asn Thr Leu Gln Pro Gly Cys Asn Ser Val Cys      50 55 60 Tyr Asp Gln Phe Phe Pro Ile Ser His Val Arg Leu Trp Ser Leu Gln  65 70 75 80 Leu Ile Leu Val Ser Thr Pro Ala Leu Leu Val Ala Met His Val Ala                  85 90 95 His Gln Gln His Ile Glu Lys Lys Met Leu Arg Leu Glu Gly His Gly             100 105 110 Asp Pro Leu His Leu Glu Glu Val Lys Arg His Lys Val His Ile Ser         115 120 125 Gly Thr Leu Trp Trp Thr Tyr Val Ile Ser Val Val Phe Arg Leu Leu     130 135 140 Phe Glu Ala Val Phe Met Tyr Val Phe Tyr Leu Leu Tyr Pro Gly Tyr 145 150 155 160 Ala Met Val Arg Leu Val Lys Cys Asp Val Tyr Pro Cys Pro Asn Thr                 165 170 175 Val Asp Cys Phe Val Ser Arg Pro Thr Glu Lys Thr Val Phe Thr Val             180 185 190 Phe Met Leu Ala Ala Ser Gly Ile Cys Ile Ile Leu Asn Val Ala Glu         195 200 205 Val Val Tyr Leu Ile Ile Arg Ala Cys Ala Arg Arg Ala Gln Arg Arg     210 215 220 Ser Asn Pro Pro Ser Arg Lys Gly Ser Gly Phe Gly His Arg Leu Ser 225 230 235 240 Pro Glu Tyr Lys Gln Asn Glu Ile Asn Lys Leu Leu Ser Glu Gln Asp                 245 250 255 Gly Ser Leu Lys Asp Ile Leu Arg Arg Ser Pro Gly Thr Gly Ala Gly             260 265 270 Leu Ala Glu Lys Ser Asp Arg Cys Ser Ala Cys         275 280

Claims (17)

A composition for diagnosing the risk of developing Sharko-Marie-Tooth in Koreans, comprising an agent for specifically detecting a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 1 or a polynucleotide comprising a complementary polynucleotide mutation site ,
The composition of the nucleotide position is the 47th nucleotide from the 5'end of SEQ ID NO: 1 G. The method according to claim 1, wherein the composition is from the 5'end of SEQ ID NO: 1 nucleotide 112 T, 139 nucleotide A, 157 nucleotide C, 414 nucleotide G, 328 nucleotide A, or a combination thereof The composition further comprises an agent for specifically detecting a polynucleotide containing a phosphorus mutation. The composition according to claim 1, wherein the agent for specifically detecting is 10 to 200 nucleotides in length. The composition according to claim 1, wherein the agent for specifically detecting is a primer, probe, or antisense nucleic acid. The composition according to claim 1, wherein the agent for specifically detecting is labeled with a detectable label. A kit for diagnosing the risk of developing Sharko-Marie-Tus in Koreans comprising the composition of claim 1 or 2. A composition for diagnosing the risk of developing Sharko-Marie-Tooth in Koreans, comprising an agent for specifically detecting a polypeptide comprising an amino acid mutation position of a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2,
The amino acid at the mutation position is a composition wherein the 16th amino acid from the N-terminal of SEQ ID NO: 2 is R. The method according to claim 7, wherein the composition from the N-terminal of SEQ ID NO: 2, the 38th amino acid is L, the 47th amino acid is K, the 53rd amino acid is R, the 138th amino acid is R, the 110th amino acid is S, or a combination thereof. A composition further comprising an agent for specifically detecting a polypeptide comprising a mutation. The composition according to claim 7, wherein the agent for specifically detecting is an antibody or an antigen-binding fragment. A kit for diagnosing the risk of developing Sharko-Marie-Tooth in Koreans comprising the composition of claim 7 or 8. A method for providing information for predicting the risk of developing Sharko-Marie-Tus in Koreans comprising the following steps:
Determining a nucleotide at position 47 from the 5'end of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 in an isolated biological sample; And
And determining that the Korean belongs to a risk group with a high probability of developing Sharko-Marie-Tooth when the 47th nucleotide from the 5'end of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 is G. . delete The method of claim 11, wherein the method comprises the steps of determining the nucleotide of the position that is 112, 139, 157, 414, 328, or a combination thereof from the 5'end of the polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1 ; And
When the 112th nucleotide from the 5'end of SEQ ID NO: 1 is T, the 139th nucleotide is A, the 157th nucleotide is C, the 414th nucleotide is G, and the 328th nucleotide is A, the Korean is characterized by the development of Sharco-Marie-Tooth. And determining that the probability belongs to a high risk group. A method for providing information for predicting the risk of developing Sharko-Marie-Tus in Koreans comprising the following steps:
Determining an amino acid at position 16 from the N-terminus of the polypeptide consisting of the amino acid sequence of SEQ ID NO: 2 in an isolated biological sample; And
And determining that the Korean belongs to a risk group having a high probability of developing Sharko-Marie-Tus when the 16th amino acid from the N-terminal of the polypeptide consisting of the amino acid sequence of SEQ ID NO: 2 is R. delete The method according to claim 14, wherein the method comprises the steps of determining the amino acid at the position of the 38th, 47th, 53th, 138th, 110th, or a combination thereof from the N-terminal of the polypeptide consisting of the amino acid sequence of SEQ ID NO:2; And
If the 38th amino acid is L, the 47th amino acid is K, the 53th amino acid is R, the 138th amino acid is R, and the 110th amino acid is S, from the N-terminal of the polypeptide consisting of the amino acid sequence of SEQ ID NO:2, Sharko- Determining that belonging to a high-risk group with a high probability of developing Marie-Tus. A method for providing information for predicting the risk of developing Sharko-Marie-Tus in Koreans comprising the following steps:
Determining an amino acid variation of the polypeptide consisting of the amino acid sequence of SEQ ID NO: 2 in the isolated biological sample; And
And determining that the Korean exhibits severe symptoms due to the development of Sharco-Marie-Tus compared to the case where the determined amino acid mutation is present in the EC domain, or in the IC domain or TM domain. .

Images