CN113637739B

CN113637739B - SCN5A mutant gene, application and Brugada syndrome detection kit

Info

Publication number: CN113637739B
Application number: CN202110906456.4A
Authority: CN
Inventors: 刘哲; 梁庆渊; 赵娜娜; 赖开生; 刘昕超; 高璇; 李方玉; 侯青; 惠汝太
Original assignee: Bestnovo Beijing Medical Technology Co Ltd
Current assignee: Baishino (Baoding) Medical Laboratory Co.,Ltd.
Priority date: 2021-08-09
Filing date: 2021-08-09
Publication date: 2022-02-01
Anticipated expiration: 2041-08-09
Also published as: CN113637739A

Abstract

The invention relates to the technical field of human genetics and cardiovascular, in particular to an SCN5A mutant gene, wherein a base C is mutated into a base T at a genome position chr3: 38592894; the reference genomic version is GRCh 37. The invention also relates to application of the SCN5A mutant gene in preparation of a Brugada syndrome detection kit and relates to the Brugada syndrome detection kit. The SCN5A mutant gene provided by the invention can distinguish Brugada syndrome patients from normal people and serve as a biomarker for clinically and auxiliarily diagnosing Brugada syndrome; the carrier for detecting the variation can provide bearing and rearing guidance and genetic counseling for the testee and reduce the birth of the infant; provides a new medicine target for the human to overcome Brugada syndrome and promotes the research and development of innovative medicines.

Description

SCN5A mutant gene, application and Brugada syndrome detection kit

Technical Field

The invention relates to the technical field of human genetics and cardiovascular, in particular to an SCN5A mutant gene, application and a Brugada syndrome detection kit.

Background

Brugada syndrome belongs to autosomal dominant inheritance, and clinical observation shows that the disease has strong familial inheritance, syncope or sudden death is often caused by ventricular fibrillation (ventricular fibrillation) or polymorphic ventricular tachycardia, and electrocardiogram right chest leads (V)_1-3) The ST segment is raised in a downward inclined shape or a saddle shape. FromBrugada syndrome received extensive attention by clinicians and researchers since its first report as a new disease species in 1992 as the brother of Brugada (Brugada syndrome, Brs).

Brugada syndrome is one of the important diseases causing idiopathic sudden ventricular fibrillation, and is very passive and passive in prevention. With the increasing discovery of genes (related to genes such as SCN5A, SCN1B, CACNA1C, GPD1-L and the like) which affect the Brugada syndrome and mutation sites, gene detection has become an important means for diagnosing the Brugada syndrome. Since the first SCN5A gene mutation was discovered by Chen (Genetic bases and molecular mechanisms for Genetic guided mutation, Nature,1998,392:293-296), a number of mutant genes related to Brugada syndrome have been reported in turn all over the world, but due to the randomness and non-tropism of gene mutation and the rareness of Brugada syndrome, the number of mutant genes discovered at present is very limited, and the discovery of any gene related to Brugada syndrome is an important technical contribution to the art.

Disclosure of Invention

The invention aims to provide an SCN5A mutant gene, application and a Brugada syndrome detection kit, wherein the provided SCN5A mutant gene can distinguish Brugada syndrome patients from normal people, is used as a biomarker for clinically and auxiliarily diagnosing Brugada syndrome, provides genetic block for families carrying Brugada syndrome pathogenic variation, and improves the quality of sound birth and sound care.

By analyzing the family members of the Brugada syndrome, the invention unexpectedly discovers that Brugada syndrome patients in the family have the following mutations:

the encoded product of the SCN5A gene is one of the subunits of tetrodotoxin-insensitive voltage-gated sodium ion channel, which is mainly expressed in the myocardium. c.4915c > T represents: in the CDS region of SCN5A gene, the 4915 th base C is mutated into base T; p.Leu1639Phe: the amino acid at position 1639 of the protein encoded by the SCN5A mutant gene changed from leucine (Leu) to phenylalanine (Phe). The CDS region of the SCN5A gene has the sequence number of SEQ ID NO. 1, and the amino acid sequence number of the SCN5A gene coding protein has the sequence number of SEQ ID NO. 2.

On the genome chr3:38592879-chr3:38592928, the sequence number of the wild type SCN5A gene is SEQ ID NO 3, and the specific sequence is GCTGCTCTTTGCCCTCATGATGTCCCTGCCTGCCCTCTTCAACATCGGGC, whereinCIs a pre-mutation base. The sequence number of the SCN5A mutant gene is SEQ ID NO. 4;

the specific sequence is GCTGCTCTTTGCCCTCATGATGTCCCTGCCTGCCTTCTTCAACATCGGGC, wherein the first and second end caps are, among others,Tis a post-mutation base.

In order to achieve the purpose, the technical scheme of the invention is as follows:

the invention provides an SCN5A mutant gene, wherein a base C is mutated into a base T at a genome position chr3: 38592894; the reference genomic version is GRCh 37.

The mutation was found to be a rare mutation by querying the population frequency database (thousand genomes: none, ESP 6500: none, ExAC: none). The results are all harmful (SIFT is 'D', Polyphen-2 is 'D', MutationTaster _ pred is 'D', VEST3 score is '0.713', and others are '5D/1H') by adopting biological information prediction software such as SIFT, Polyphen-2 and the like for cross prediction, and the amino acid change caused by the mutation is likely to influence the protein function. The amino acid is changed from leucine to phenylalanine. The database was queried to find that the amino acid at this position was highly conserved in vertebrates. Querying ClinVar and HGMD databases to find no variation, and evaluating missense variation p.Met1633Val near the site as suspected pathogenic mutation by a reporter (NM-198056.2, ClinVar database, related diseases: not provided); the literature search does not find the mutation and disease related report. According to the existing evidence: the mutation is a rare variation, the software predicts that the variation may have influence on the protein function, the amino acid at the position is highly conserved in vertebrates, but family linkage and functional evidence support are lacked, so the variation is suspicious pathogenic variation.

The invention also provides application of the SCN5A mutant gene in preparing a Brugada syndrome detection kit.

The invention also provides a Brugada syndrome detection kit, which comprises a reagent for detecting human SCN5A mutant genes, wherein the base C is mutated into the base T at the genomic position chr3: 38592894.

Preferably, primer sequences SEQ ID NO 5 and SEQ ID NO 6 are also included.

Fourthly, the beneficial effects of the invention are as follows:

the hybrid missense mutation of the base C to the base T (namely the human SCN5A gene c.4915C > T hybrid missense mutation) at the genomic position chr3:38592894 can distinguish Brugada syndrome patients from normal people and serve as a biomarker for clinically and auxiliarily diagnosing Brugada syndrome. Whether the subject carries the variation or not is detected, and the carrier of the variation is detected, so that prenatal and postnatal care guidance and genetic counseling can be provided for the subject, and the birth of the infant patient is reduced; provides a new medicine target for the human to overcome Brugada syndrome and promotes the research and development of innovative medicines.

Drawings

FIG. 1 is a graph of Brugada syndrome family;

FIG. 2 is a Sanger sequencing graph of probands in a family;

figure 3 is a Sanger sequencing plot of non-diseased members of the pedigree.

Detailed Description

The following is further detailed by way of specific embodiments:

example 1 proband verification experiment

Sample source: the Fujian medical university attaches to the first hospital, sends 5-10mL of whole blood sample under the premise that the proband (36 years old) and the family voluntarily sign informed consent, establishes a medical record database, and records the information of the disease condition, family condition and the like of the proband in detail. The study was approved by the ethical committee of the unit.

Clinical profile of proband:

TABLE 1 clinical profiles of probands

The Sanger sequencing method is used for detecting the c.4915C > T heterozygous missense variation of the human SCN5A gene, and specifically comprises the following steps:

s1, extracting whole genome DNA:

the whole genome DNA extraction kit of the magnetic bead method whole genome DNA extraction kit of Jiangsu Baishinuo medical science and technology Limited company is adopted to extract the whole genome DNA of the anticoagulated human whole blood EDTA sample of the proband and the verification sample, and the concentration and the purity of the DNA are detected.

S2, amplifying the SCN5A gene by using a primer;

(1) the PCR amplification reagents were prepared, and the compositions of the PCR amplification reagents are specifically shown in the following Table 2:

TABLE 2 composition of PCR amplification reagents

2 × Taq MasterMix (Dye) contains the following components: taq DNA Polymerase, PCR Buffer, Mg² ⁺dNTPs, PCR stabilizer and enhancer, etc. required by conventional PCR.

The primer information required for amplification is shown in table 3 below:

TABLE 3 primer information

Sequence numbering	Primer name	Sequence (5 '→ 3')
			SEQ ID NO:5	Forward primer (SCN5A-E27-PART-F2)	GCTCCTTGCCATATAGAGACCC
SEQ ID NO:6	Reverse primer (SCN5A-E27-PART-R2)	AACATATCGAAGTCGTCCTCAC

(2) Amplification of a fragment of interest

The reaction systems were mixed, and amplification reaction of the target gene fragment was performed on a PCR instrument, and the amplification procedure was as shown in table 3 below. After amplification was completed, the PCR product was purified using Agencourt AMPure XP magnetic beads (purchased from beckman coulter trade (china) ltd.). The PCR amplification procedure was: pre-denaturation at 94 ℃ for 2 min; denaturation at 94 ℃ for 30s, annealing at 61 ℃ for 30s, and extension at 72 ℃ for 30s for 30 cycles. Final extension at 72 ℃ for 2 min.

2 mu L of PCR product is taken, 1.5% agarose gel electrophoresis is used for detecting the PCR product, and 1000bp Marker is selected as reference.

S3, Sanger sequencing of the PCR products using a 3730XL Genetic Analyzer full-automatic sequencer, obtaining reference sequences from NCBI (https:// www.ncbi.nlm.nih.gov /) database and aligning the sequencing results.

The results as shown in fig. 1-3 indicate that: probands and their children carry the SCN5A gene c.4915c > T heterozygous missense variation inherited from the mother of probands. Wherein, FIG. 1 is a Brugada syndrome family diagram; FIG. 2 is a Sanger sequencing graph of probands in a family; figure 3 is a Sanger sequencing plot of non-diseased members of the pedigree.

Example 2-detection kit for Brugada syndrome

This example provides a kit for detecting human SCN5A gene c.4915c > T heterozygous missense variation, comprising 2 × Taq MasterMix (Dye), primers capable of detecting SCN5A mutant gene, etc., and the specific composition of the kit is shown in table 4 below.

TABLE 4 kit composition

The specific steps of screening the Brugada syndrome by using the kit are as follows: the DNA of the subject was extracted according to the procedure of example 1, and then the SCN5A gene was amplified using the designed primer combination (SEQ ID NO:5 and SEQ ID NO:6) to obtain a PCR product, and finally the PCR product was sequenced. And obtaining a reference sequence from an NCBI (https:// www.ncbi.nlm.nih.gov /) database, comparing the reference sequence with a sequencing result, judging whether the SCN5A gene of the testee carries c.4915C > T heterozygous missense variation, and assisting the clinical confirmation of whether the testee has Brugada syndrome.

Example 3 mutation verification against out-of-family Normal persons

The site of the mutation in SCN5A gene c.4915C > T was determined in 500 non-related normal persons of the same race (i.e., non-familial normal persons) by the method described in example 1, and none of the results showed that the mutation could be detected.

Taken together, and based on the fact that the SCN5A gene c.4915c > T heterozygous missense mutation results in p.leu1639phe change of the protein encoded by the SCN5A gene, while the SCN5A gene is a known pathogenic gene of Brugada syndrome, the SCN5A gene c.4915c > T heterozygous missense mutation is proved to be a pathogenic mutation of Brugada syndrome.

The foregoing detailed description of the preferred embodiments of the invention has been presented. It should be understood that numerous modifications and variations could be devised by those skilled in the art in light of the present teachings without departing from the inventive concepts. Therefore, the technical solutions available to those skilled in the art through logic analysis, reasoning and limited experiments based on the prior art according to the concept of the present invention should be within the scope of protection defined by the claims.

Sequence listing

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Pro Asp Leu Tyr Gly Asn Pro Pro Gln Glu Leu Ile Gly Glu Pro Leu

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Glu Asp Leu Asp Pro Phe Tyr Ser Thr Gln Lys Thr Phe Ile Val Leu

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Val Leu Ser Pro Phe His Pro Ile Arg Arg Ala Ala Val Lys Ile Leu

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Trp Asn Trp Leu Asp Phe Ser Val Ile Ile Met Ala Tyr Val Ser Glu

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Met Gly Asn Leu Arg His Lys Cys Val Arg Asn Phe Thr Ala Leu Asn

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Gly Thr Asn Gly Ser Val Glu Ala Asp Gly Leu Val Trp Glu Ser Leu

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Asp Leu Tyr Leu Ser Asp Pro Glu Asn Tyr Leu Leu Lys Asn Gly Thr

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Ser Asp Val Leu Leu Cys Gly Asn Ser Ser Asp Ala Gly Thr Cys Pro

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Glu Gly Tyr Arg Cys Leu Lys Ala Gly Glu Asn Pro Asp His Gly Tyr

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Thr Ser Phe Asp Ser Phe Ala Trp Ala Phe Leu Ala His Phe Arg Leu

355 360 365

Met Ala Gln Asp Cys Trp Glu Arg Leu Tyr Gln Gln Thr Leu Arg Ser

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Ala Gly Lys Ile Tyr Met Ile Phe Phe Met Leu Val Ile Phe Leu Gly

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Ser Phe Tyr Leu Val Asn Leu Ile Leu Ala Val Val Ala Met Ala Tyr

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Glu Glu Gln Asn Gln Ala Thr Ile Ala Glu Thr Glu Glu Lys Glu Lys

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Arg Phe Gln Glu Ala Met Glu Met Leu Lys Lys Glu His Glu Ala Leu

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Thr Ile Arg Gly Val Asp Thr Val Ser Arg Ser Ser Leu Glu Met Ser

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Pro Leu Ala Pro Val Asn Ser His Glu Arg Arg Ser Lys Arg Arg Lys

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Arg Met Ser Ser Gly Thr Glu Glu Cys Gly Glu Asp Arg Leu Pro Lys

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Ser Asp Ser Glu Asp Gly Pro Arg Ala Met Asn His Leu Ser Leu Thr

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Arg Gly Leu Ser Arg Thr Ser Met Lys Pro Arg Ser Ser Arg Gly Ser

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Ile Phe Thr Phe Arg Arg Arg Asp Leu Gly Ser Glu Ala Asp Phe Ala

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Asp Asp Glu Asn Ser Thr Ala Gly Glu Ser Glu Ser His His Thr Ser

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Leu Leu Val Pro Trp Pro Leu Arg Arg Thr Ser Ala Gln Gly Gln Pro

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Ser Pro Gly Thr Ser Ala Pro Gly His Ala Leu His Gly Lys Lys Asn

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Ser Thr Val Asp Cys Asn Gly Val Val Ser Leu Leu Gly Ala Gly Asp

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Pro Glu Ala Thr Ser Pro Gly Ser His Leu Leu Arg Pro Val Met Leu

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Glu His Pro Pro Asp Thr Thr Thr Pro Ser Glu Glu Pro Gly Gly Pro

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Gln Met Leu Thr Ser Gln Ala Pro Cys Val Asp Gly Phe Glu Glu Pro

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Gly Ala Arg Gln Arg Ala Leu Ser Ala Val Ser Val Leu Thr Ser Ala

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Leu Glu Glu Leu Glu Glu Ser Arg His Lys Cys Pro Pro Cys Trp Asn

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Arg Leu Ala Gln Arg Tyr Leu Ile Trp Glu Cys Cys Pro Leu Trp Met

690 695 700

Ser Ile Lys Gln Gly Val Lys Leu Val Val Met Asp Pro Phe Thr Asp

705 710 715 720

Leu Thr Ile Thr Met Cys Ile Val Leu Asn Thr Leu Phe Met Ala Leu

725 730 735

Glu His Tyr Asn Met Thr Ser Glu Phe Glu Glu Met Leu Gln Val Gly

740 745 750

Asn Leu Val Phe Thr Gly Ile Phe Thr Ala Glu Met Thr Ser Lys Ile

755 760 765

Ile Ala Leu Asp Pro Tyr Tyr Tyr Phe Gln Gln Gly Trp Asn Ile Phe

770 775 780

Asp Ser Ile Ile Val Ile Leu Ser Leu Met Glu Leu Gly Leu Ser Arg

785 790 795 800

Met Ser Asn Leu Ser Val Leu Arg Ser Phe Arg Leu Leu Arg Val Phe

805 810 815

Lys Leu Ala Lys Ser Trp Pro Thr Leu Asn Thr Leu Ile Lys Ile Ile

820 825 830

Gly Asn Ser Val Gly Ala Leu Gly Asn Leu Thr Leu Val Leu Ala Ile

835 840 845

Ile Val Phe Ile Phe Ala Val Val Gly Met Gln Leu Phe Gly Lys Asn

850 855 860

Tyr Ser Glu Leu Arg Asp Ser Asp Ser Gly Leu Leu Pro Arg Trp His

865 870 875 880

Met Met Asp Phe Phe His Ala Phe Leu Ile Ile Phe Arg Ile Leu Cys

885 890 895

Gly Glu Trp Ile Glu Thr Met Trp Asp Cys Met Glu Val Ser Gly Gln

900 905 910

Ser Leu Cys Leu Leu Val Phe Leu Leu Val Met Val Ile Gly Asn Leu

915 920 925

Val Val Leu Asn Leu Phe Leu Ala Leu Leu Arg Ser Ser Phe Ser Ala

930 935 940

Asp Asn Leu Thr Ala Pro Asp Glu Asp Arg Glu Met Asn Asn Leu Gln

945 950 955 960

Leu Ala Leu Ala Arg Ile Gln Arg Gly Leu Arg Phe Val Lys Arg Thr

965 970 975

Thr Trp Asp Phe Cys Cys Gly Leu Leu Arg Gln Arg Pro Gln Lys Pro

980 985 990

Ala Ala Leu Ala Ala Gln Gly Gln Leu Pro Ser Cys Ile Ala Thr Pro

995 1000 1005

Tyr Ser Pro Pro Pro Pro Glu Thr Glu Lys Val Pro Pro Thr Arg Lys

1010 1015 1020

Glu Thr Arg Phe Glu Glu Gly Glu Gln Pro Gly Gln Gly Thr Pro Gly

1025 1030 1035 1040

Asp Pro Glu Pro Val Cys Val Pro Ile Ala Val Ala Glu Ser Asp Thr

1045 1050 1055

Asp Asp Gln Glu Glu Asp Glu Glu Asn Ser Leu Gly Thr Glu Glu Glu

1060 1065 1070

Ser Ser Lys Gln Glu Ser Gln Pro Val Ser Gly Gly Pro Glu Ala Pro

1075 1080 1085

Pro Asp Ser Arg Thr Trp Ser Gln Val Ser Ala Thr Ala Ser Ser Glu

1090 1095 1100

Ala Glu Ala Ser Ala Ser Gln Ala Asp Trp Arg Gln Gln Trp Lys Ala

1105 1110 1115 1120

Glu Pro Gln Ala Pro Gly Cys Gly Glu Thr Pro Glu Asp Ser Cys Ser

1125 1130 1135

Glu Gly Ser Thr Ala Asp Met Thr Asn Thr Ala Glu Leu Leu Glu Gln

1140 1145 1150

Ile Pro Asp Leu Gly Gln Asp Val Lys Asp Pro Glu Asp Cys Phe Thr

1155 1160 1165

Glu Gly Cys Val Arg Arg Cys Pro Cys Cys Ala Val Asp Thr Thr Gln

1170 1175 1180

Ala Pro Gly Lys Val Trp Trp Arg Leu Arg Lys Thr Cys Tyr His Ile

1185 1190 1195 1200

Val Glu His Ser Trp Phe Glu Thr Phe Ile Ile Phe Met Ile Leu Leu

1205 1210 1215

Ser Ser Gly Ala Leu Ala Phe Glu Asp Ile Tyr Leu Glu Glu Arg Lys

1220 1225 1230

Thr Ile Lys Val Leu Leu Glu Tyr Ala Asp Lys Met Phe Thr Tyr Val

1235 1240 1245

Phe Val Leu Glu Met Leu Leu Lys Trp Val Ala Tyr Gly Phe Lys Lys

1250 1255 1260

Tyr Phe Thr Asn Ala Trp Cys Trp Leu Asp Phe Leu Ile Val Asp Val

1265 1270 1275 1280

Ser Leu Val Ser Leu Val Ala Asn Thr Leu Gly Phe Ala Glu Met Gly

1285 1290 1295

Pro Ile Lys Ser Leu Arg Thr Leu Arg Ala Leu Arg Pro Leu Arg Ala

1300 1305 1310

Leu Ser Arg Phe Glu Gly Met Arg Val Val Val Asn Ala Leu Val Gly

1315 1320 1325

Ala Ile Pro Ser Ile Met Thr Val Leu Leu Val Cys Leu Ile Phe Trp

1330 1335 1340

Leu Ile Phe Ser Ile Met Gly Val Asn Leu Phe Ala Gly Lys Phe Gly

1345 1350 1355 1360

Arg Cys Ile Asn Gln Thr Glu Gly Asp Leu Pro Leu Asn Tyr Thr Ile

1365 1370 1375

Val Asn Asn Lys Ser Gln Cys Glu Ser Leu Asn Leu Thr Gly Glu Leu

1380 1385 1390

Tyr Trp Thr Lys Val Lys Val Asn Phe Asp Asn Val Gly Ala Gly Tyr

1395 1400 1405

Leu Ala Leu Leu Gln Val Tyr Glu Glu Gln Pro Gln Trp Glu Tyr Asn

1410 1415 1420

Leu Tyr Met Tyr Ile Tyr Phe Val Ile Phe Ile Ile Phe Gly Ser Phe

1425 1430 1435 1440

Phe Thr Leu Asn Leu Phe Ile Gly Val Ile Ile Asp Asn Phe Asn Gln

1445 1450 1455

Gln Lys Lys Lys Leu Gly Gly Gln Asp Ile Phe Met Thr Glu Glu Gln

1460 1465 1470

Lys Lys Tyr Tyr Asn Ala Met Lys Lys Leu Gly Ser Lys Lys Pro Gln

1475 1480 1485

Lys Pro Ile Pro Arg Pro Leu Asn Lys Tyr Gln Gly Phe Ile Phe Asp

1490 1495 1500

Ile Gly Thr Lys Gln Ala Phe Asp Val Thr Ile Met Phe Leu Ile Cys

1505 1510 1515 1520

Leu Asn Met Val Thr Met Met Val Glu Thr Asp Asp Gln Ser Pro Glu

1525 1530 1535

Lys Ile Asn Ile Leu Ala Lys Ile Asn Leu Leu Phe Val Ala Ile Phe

1540 1545 1550

Thr Gly Glu Cys Ile Val Lys Leu Ala Ala Leu Arg His Tyr Tyr Phe

1555 1560 1565

Thr Asn Ser Trp Asn Ile Phe Asp Phe Val Val Val Ile Leu Ser Ile

1570 1575 1580

Val Gly Thr Val Leu Ser Asp Ile Ile Gln Lys Tyr Phe Phe Ser Pro

1585 1590 1595 1600

Thr Leu Phe Arg Val Ile Arg Leu Ala Arg Ile Gly Arg Ile Leu Arg

1605 1610 1615

Leu Ile Arg Gly Ala Lys Gly Ile Arg Thr Leu Leu Phe Ala Leu Met

1620 1625 1630

Met Ser Leu Pro Ala Leu Phe Asn Ile Gly Leu Leu Leu Phe Leu Val

1635 1640 1645

Met Phe Ile Asp Ser Ile Phe Gly Met Ala Asn Phe Ala Tyr Val Lys

1650 1655 1660

Trp Glu Ala Gly Ile Asp Asp Met Phe Asn Phe Gln Thr Phe Ala Asn

1665 1670 1675 1680

Asn Met Leu Cys Leu Phe Gln Ile Thr Thr Ser Ala Gly Trp Asp Gly

1685 1690 1695

Leu Leu Ser Pro Ile Leu Asn Thr Gly Pro Pro Tyr Cys Asp Pro Thr

1700 1705 1710

Leu Pro Asn Ser Asn Gly Ser Arg Gly Asp Cys Gly Ser Pro Ala Val

1715 1720 1725

Gly Ile Leu Phe Phe Thr Thr Tyr Ile Ile Ile Ser Phe Leu Ile Val

1730 1735 1740

Val Asn Met Tyr Ile Ala Ile Ile Leu Glu Asn Phe Ser Val Ala Thr

1745 1750 1755 1760

Glu Glu Ser Thr Glu Pro Leu Ser Glu Asp Asp Phe Asp Met Phe Tyr

1765 1770 1775

Glu Ile Trp Glu Lys Phe Asp Pro Glu Ala Thr Gln Phe Ile Glu Tyr

1780 1785 1790

Ser Val Leu Ser Asp Phe Ala Asp Ala Leu Ser Glu Pro Leu Arg Ile

1795 1800 1805

Ala Lys Pro Asn Gln Ile Ser Leu Ile Asn Met Asp Leu Pro Met Val

1810 1815 1820

Ser Gly Asp Arg Ile His Cys Met Asp Ile Leu Phe Ala Leu Thr Lys

1825 1830 1835 1840

Arg Val Leu Gly Glu Ser Gly Glu Met Asp Ala Leu Lys Ile Gln Met

1845 1850 1855

Glu Glu Lys Phe Met Ala Ala Asn Pro Ser Lys Ile Ser Tyr Glu Pro

1860 1865 1870

Ile Thr Thr Thr Leu Arg Arg Lys His Glu Glu Val Ser Ala Met Val

1875 1880 1885

Ile Gln Arg Ala Phe Arg Arg His Leu Leu Gln Arg Ser Leu Lys His

1890 1895 1900

Ala Ser Phe Leu Phe Arg Gln Gln Ala Gly Ser Gly Leu Ser Glu Glu

1905 1910 1915 1920

Asp Ala Pro Glu Arg Glu Gly Leu Ile Ala Tyr Val Met Ser Glu Asn

1925 1930 1935

Phe Ser Arg Pro Leu Gly Pro Pro Ser Ser Ser Ser Ile Ser Ser Thr

1940 1945 1950

Ser Phe Pro Pro Ser Tyr Asp Ser Val Thr Arg Ala Thr Ser Asp Asn

1955 1960 1965

Leu Gln Val Arg Gly Ser Asp Tyr Ser His Ser Glu Asp Leu Ala Asp

1970 1975 1980

Phe Pro Pro Ser Pro Asp Arg Asp Arg Glu Ser Ile Val

1985 1990 1995

<210> 3

<211> 50

<212> DNA

<213> Homo sapiens

<400> 3

gctgctcttt gccctcatga tgtccctgcc tgccctcttc aacatcgggc 50

<210> 4

<211> 50

<212> DNA

<213> Homo sapiens

<400> 4

gctgctcttt gccctcatga tgtccctgcc tgccttcttc aacatcgggc 50

<210> 5

<211> 22

<212> DNA

<213> Homo sapiens

<400> 5

gctccttgcc atatagagac cc 22

<210> 6

<211> 22

<212> DNA

<213> Homo sapiens

<400> 6

aacatatcga agtcgtcctc ac 22

Claims

1. The SCN5A mutant gene is characterized in that a base C is mutated into a base T at a genomic position chr3: 38592894; the reference genomic version is GRCh 37.

2. The SCN5A mutant gene according to claim 1, wherein the amino acid change of the protein encoded by the SCN5A mutant gene is: leu is leucine and Phe is phenylalanine.

3. Use of a reagent for detecting the SCN5A mutant gene of claim 1 or 2 in the preparation of a kit.

4. A Brugada syndrome detection kit is characterized by comprising a reagent for detecting human SCN5A mutant genes, wherein at the genomic position chr3:38592894, a base C is mutated into a base T; the reference genomic version is GRCh 37.

5. The Brugada syndrome assay kit of claim 4, further comprising primer sequences SEQ ID NO 5 and SEQ ID NO 6.