CN109022566B

CN109022566B - Gene mutation target related to paroxysmal movement-induced dyskinesia and application thereof

Info

Publication number: CN109022566B
Application number: CN201810778601.3A
Authority: CN
Inventors: 田沃土; 曹立
Original assignee: Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Current assignee: Ruinjin Hospital Affiliated to Shanghai Jiaotong University School of Medicine Co Ltd
Priority date: 2018-07-16
Filing date: 2018-07-16
Publication date: 2022-04-05
Anticipated expiration: 2038-07-16
Also published as: CN109022566A

Abstract

The invention relates to the technical field of medical biological detection, in particular to gene mutation targets related to paroxysmal movement-induced dyskinesia and application of the gene mutation targets in a reagent or a kit for diagnosing or treating the paroxysmal movement-induced dyskinesia and evaluating the prognosis of the paroxysmal movement-induced dyskinesia. The invention discovers mutation targets c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T of PRRT2 genes which are main pathogenic genes of paroxysmal movement-induced dyskinesia, can be effectively used for development judgment, treatment scheme selection and/or prognosis evaluation of the paroxysmal movement-induced dyskinesia, and has a certain clinical application prospect.

Description

Gene mutation target related to paroxysmal movement-induced dyskinesia and application thereof

Technical Field

The invention relates to the technical field of medical biological detection, and relates to gene mutations related to paroxysmal movement-induced dyskinesia, and applications of the gene mutations in a reagent or a kit for diagnosing or treating the paroxysmal movement-induced dyskinesia and evaluating the prognosis of the paroxysmal movement-induced dyskinesia.

Background

Paroxysmal dyskinesia (PKD) is a recurrent involuntary disorder of the involuntary movement, the most common subtype of Paroxysmal dyskinesia (PxDs). It is manifested by involuntary movements or abnormal postures of limbs or trunk induced from rest to sudden voluntary movements, and may be manifested by dystonia, chorea-like athetosis or tossing-like movements, with short duration of a single attack (several seconds to 1 minute), no pain and loss of consciousness during the attack, and return to normal during the attack.

PKD is more sick in young children or adolescents, and male is more sick than female; the onset of the disease is induced by a definite inducing factor, i.e. the disease occurs from rest to sudden movement, such as rising, stepping, running, changing the exercise speed, etc., or by fright, etc., and the symptoms may be aggravated under the state of mental stress or sleep insufficiency; the attack duration is short and is not more than 60 seconds, and the attack frequency of a patient is different from several times to hundreds of times every day; the frequency of the attacks is correlated with age, i.e., high peaks of the attacks generally occur before and after the puberty.

Depending on its etiologic classification, PKD can be classified as genetic, sporadic, or secondary, with primary PKD being more common. The secondary PKD can be secondary to various nervous system diseases, such as cerebral apoplexy, multiple sclerosis, cerebral trauma, perinatal cerebral anoxia, central nervous system infection and the like, and can also be secondary to some endocrine or metabolic disorder diseases, such as hyperthyroidism, primary hypoparathyroidism, diabetes, hypoglycemia and the like. The hereditary PKD is mainly inherited in autosomal dominant way and is accompanied by incomplete exon phenomenon.

A total of 3 PKD-associated sites were currently found, namely EKD 1-3. In 2011, PRRT2(Proline-rich transmembrane protein 2) was first confirmed by the inventor and other domestic research groups as the causative gene of PKD. At present, the incidence of PRRT2 gene mutation in PKD cases reported worldwide is about 35-73%, suggesting that PRRT2 is the main causative gene of PKD.

Genetic diagnosis and prenatal and postnatal care of PKD patients have been a problem and a research target of considerable interest in the fields of neuroscience and genetics. Meanwhile, the efficient gene detection technology and the definite genetic diagnosis are helpful for the diagnosis of PKD patients and the selection of reasonable treatment schemes.

However, the pathogenesis of PKD diseases caused by PRRT2 gene mutation is not clear, the pathogenic mutation sites are more, and the phenotypic-genotypic heterogeneity of PRRT2 gene is always a research hotspot in the fields of neuroscience and clinical medicine. To date, the phenotype-genotype correlation of PKD has not been elucidated. This is a major problem, and one of the important reasons is that the pathogenesis of PKD may involve multiple links and central loops, such as neurotransmitter release, ion channel abnormalities, and neuronal excitability abnormalities.

Therefore, it is expected that if only a few known PRRT2 hotspot mutations are detected in a one-generation sequencing test for PKD patients, the result will have a certain negative probability for all PKD patients by using the same gene diagnosis strategy, and the establishment of future health care treatment for patients with negative detection results will bring about great trouble. Therefore, the most effective method is to find out the mutation situation of PRRT2 gene of each PKD patient by first-generation sequencing, then analyze the coseparation situation in the family according to the specific gene mutation, further evaluate the pathogenicity of the mutation, and establish a personalized diagnosis and treatment scheme and a sound birth and sound care strategy.

Therefore, it is important to find the mutation site of the main pathogenic gene of paroxysmal movement-induced dyskinesia and further understand the biological function of the mutation site in the early stage lesion of paroxysmal movement-induced dyskinesia. However, at present, the mutation sites of the PRRT2 gene, which is the main pathogenic gene, have not been studied in detail aiming at a large number of samples of patients with paroxysmal movement-induced dyskinesia, and new mutation sites with strong pathogenicity have not been found, and no relevant literature reports exist.

Disclosure of Invention

The present invention has been made to solve the above problems, and an object of the present invention is to provide a diagnostic marker for paroxysmal dyskinesia, and an object of the present invention is also to provide a novel use of a target of a gene mutation associated with paroxysmal dyskinesia, that is, an application in the preparation of a kit for diagnosing and prognostically evaluating paroxysmal dyskinesia.

According to a first aspect of the present invention, there is provided a paroxysmal movement-induced dyskinesia-associated gene mutation target, which is c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene.

The invention extracts the whole genome from the peripheral blood samples of 284 cases of paroxysmal motor-induced dyskinesia patients and carries out exon sequencing, which is the sequencing research of the largest sample of paroxysmal motor-induced dyskinesia at home and abroad at present, so that the treatment and sound-birth and sound-care scheme for the paroxysmal motor-induced dyskinesia patients is formulated according to the mutant gene target points found in the research, and the scheme is definitely more targeted and more reasonable.

Therefore, the present invention intends to provide a method for clinical treatment and patient's sound birth and care based entirely on the gene sequencing results of patients with paroxysmal movement-induced dyskinesia, so-called personalized diagnosis and treatment scheme.

The personalized diagnosis and treatment scheme provided by the invention is mainly based on two factors, namely the sequencing result of the PRRT2 gene exon in the peripheral blood genome of each patient, and the actual age and physical condition of each patient.

In 284 patients with paroxysmal movement-induced dyskinesia studied, 4 new pathogenic mutations of PRRT2 gene were co-found, which could lead to the development of the disease, respectively, c.1012+5g > a, c.959c > T (p.a320v), c.971_972insG (p.v325sfs × 16) and c.761c > T (p.p254l). Among them, c.959c > T (p.a320v) was found in one sporadic PKD case; three additional gene mutations c.1012+5g > a, c.971_972insG (p.v325sfs 16) and c.761c > T (p.p254l) were co-isolated in three PKD families, respectively.

The mutation c.959c > T (p.a320v) was found in a sporadic PKD patient, a 25 year old male, who developed involuntary bilateral limb writhing and chorea-like symptoms in sudden movements or changes in movement speed from age 8, with a single episode lasting about 20 seconds with facial involvement, speech deficit and fluent speech, with 2-3 episodes per day with pre-felt prior to each episode, and oral carbamazepine (150 mg/day) allowed complete control of symptoms, consistent with clinical diagnostic criteria for PKD.

Wherein c.761C > T (p.P254L) exists in a 39-year-old male patient, the patient has involuntary movements of bilateral limbs when suddenly moving from 17 years old, the involuntary movements are expressed as inversion of feet and pronation of upper limbs, the single attack time is less than 10 seconds, the oral carbamazepine (100 mg/day) can effectively control symptoms, the clinical characteristics of PKD are met, and similar clinical manifestations exist in brother of the patient, and the same mutation also exists.

In addition, the mutation c.971_972insG (p.v325sfs 16) was co-isolated in one PKD family. The patient is a 9 year old infant, and has involuntary movement during sudden movement (such as getting up and walking, passing traffic lights, etc.), manifested by stiff limbs or involuntary wriggling accompanied by stiff facial expression, single attack lasting less than 30 seconds, pre-sensation before each attack, manifested by numbness or stiffness of upper limbs from one side, and attack 5-6 times per day. The infant suffers from twice infant convulsions within 1 year of age. The infant mother also has a history of similar symptom onset between 9-30 years of age.

The PRRT2 gene cleavage site mutation c.1012+5g > a was co-isolated in another familial PKD family. In the family, the proband is a female in the age of 10 years, starting from the age of 9 years, when playing badminton or running for 50 meters, the left limb involuntary movement appears, the left limb involuntary movement is expressed as dystonia and chorea-like movement, the face is involved, the expression stiffness and dysarthria are expressed, the duration of a single attack is less than 1 minute, and the attack frequency is 10 times/day. Their mothers also had similar symptoms occurring from age 10 and were found to have the same mutation c.1012+5g > a.

These 4 mutations were scored as pathogenic mutations, evaluated according to ACMG guidelines and standards. The PRRT2 gene contains a total of 4 exons and encodes a proline-rich transmembrane protein 2(PRRT 2). According to a Human Gene Mutation Database (HGMD), by 6 months of 2018, 92 common PRRT2 Gene mutations are reported, wherein common Mutation types comprise nonsense Mutation, missense Mutation, insertion Mutation, shearing Mutation and The like, and in addition, micro deletion/insertion, large fragment deletion/insertion, compound heterozygosis, homozygous Mutation and The like exist; the vast majority are located in

exons

2 and 3 of the gene.

In a second aspect of the invention, the application of the gene mutation target as a diagnostic marker of paroxysmal movement-induced dyskinesia is provided.

The third aspect of the invention provides the application of the gene mutation target point in preparing a reagent or a kit for diagnosing and prognostically evaluating paroxysmal movement-induced dyskinesia.

The diagnostic and prognostic evaluation reagent is a reagent for detecting c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene in a biological sample, and the kit comprises a reagent for detecting c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene in the biological sample.

Preferably, the reagent for detecting the c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene in the biological sample is selected from: PCR primers with detection specificity for PRRT2 gene.

The PCR primer sequence with detection specificity to PRRT2 gene is as follows:

the biological sample is selected from: peripheral blood obtained from a subject.

In a fourth aspect of the present invention, there is provided a diagnostic and prognostic assessment kit for paroxysmal movement-induced dyskinesia, which comprises reagents for detecting the c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene in a biological sample.

Preferably, the diagnostic kit comprises: (i) one or more reagents for detecting an effective amount of the c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites; (ii) one or more selected from the group consisting of: containers, instructions for use, positive control sequences, negative control sequences, buffers, adjuvants or solvents, such as solutions for suspending or immobilizing cells, solutions for lysing cells, or solutions for nucleic acid purification.

Wherein, the positive control sequence is a gene amplification sequence of a PKD patient known to be caused by PRRT2 mutation, and the negative control sequence is a gDNA sequence (NM-145239) of a GRCH38/hg38 human standard PRRT2 gene.

The kit of the present invention may further include instructions for use of the kit, wherein the instructions describe a specific procedure for detection using the kit, and a method for evaluating the degree of a disease state of paroxysmal exercise-induced dyskinesia and a strategy for sound prenatal and postnatal care using the detection results.

With the diagnostic and prognostic assessment kits of the present invention, PRRT2 molecules can be detected by various methods selected from the group consisting of (including but not limited to): performing Polymerase Chain Reaction (PCR) or biochip detection method on the target fragment. The detection mode can be adjusted and changed by those skilled in the art according to actual conditions and needs.

Of course, other reagents that are clinically useful in the determination of outcome of a patient's paroxysmal exercise-induced dyskinesia, selection of treatment regimens, and/or prognostic assessment are also included in the kits to assist or validate the results obtained by detecting the PRRT2 molecule.

In a fifth aspect of the present invention, there is provided a method for detecting paroxysmal movement-induced dyskinesia using the above kit, comprising:

A. obtaining a peripheral blood sample from an object, centrifuging the blood sample to be detected at normal temperature to obtain plasma, extracting peripheral blood genome DNA by using a reagent provided in a reagent kit, and determining the purity of the plasma;

B. and (3) carrying out quantitative detection on the copy number of the target fragment by adopting a real-time quantitative PCR technology, wherein primers used in the detection process are shown as SEQ ID NO. 1-8.

C. Sending the PCR product to a first-generation sequencing detection, and comparing the target gene with a normal control sequence;

D. according to the detection result, carrying out coseparation verification on the same mutation on other members in the patient family: if the detection result is consistent with the diseased condition, the detected mutation is suggested to be a pathogenic mutation.

In the present invention, the term "control level" refers to the gDNA sequence of the normal population of PRRT2 molecules used as a reference.

The term "prognosis" refers to predicting the likely course and outcome of a disease, which includes determining the specific outcome of the disease (e.g., improvement, the appearance or disappearance of other abnormalities such as certain symptoms, signs, and complications). Poor prognosis as described in the present invention includes, but is not limited to: no obvious tendency to self-heal after adulthood, more frequent attacks and longer duration of single attacks in the case of taking larger doses of carbamazepine or oxcarbazepine, etc. After predicting the prognosis of the patient, a more rational treatment can be selected to improve the patient's prognosis by combining with the specific mutation site of the PRRT2 molecule.

The invention has the following beneficial guarantee and effects:

the invention is based on the whole genome exon sequencing result of the paroxysmal movement-induced dyskinesia patients, namely, the corresponding clinical diagnosis and treatment scheme adopted according to the exon mutation condition of the patients is beneficial to evaluating the state of illness and is more beneficial to selecting and evaluating a treatment method. Because the clinical treatment scheme and the method provided by the invention are formulated based on the big data analysis result of the whole genome exon detection of the patient, the adopted clinical treatment scheme is more individualized, has more pertinence and targeting property than other methods, and has better treatment effect.

The present inventors have discovered 4 novel mutations of the unprompted PRRT2 gene that can lead to paroxysmal movement-induced dyskinesia, including c.1012+5g > a, c.959c > T (p.a320v), c.971_972insG (p.v325sfs 16) and c.761c > T (p.p254l). The gene mutation targets can be effectively used for genetic detection, treatment scheme selection and/or prognosis evaluation of paroxysmal movement-induced dyskinesia, so that a novel diagnosis agent and/or detection agent for paroxysmal movement-induced dyskinesia genes is provided for the field, and the gene mutation targets have certain clinical application prospects.

In addition, in terms of technology, the detection of gene mutation targets is essentially quantitative PCR detection of blood genomes, has the characteristics of simple and convenient operation, sensitive detection, good specificity, high repeatability and the like, and is increasingly applied to clinical examination technology nowadays.

In addition, the index gene mutation target sequence related by the invention has obvious difference with a normal target sequence, has higher clinical reference value and reliability, can improve the detection rate and accuracy of the familial paroxysmal motion-induced dyskinesia, has great significance for the clinical treatment of the familial paroxysmal motion-induced dyskinesia, can obtain a detection result only by collecting blood of a detector, and has high compliance of patients.

Drawings

FIG. 1 is a family diagram of four mutant targets of PRRT2 pathogenic gene. A: the PRRT2 mutation c.761C > T (p.P254L) was detected in pedigree A, present in proband and Cologe (II:2 and II:1), but absent from normal pedigree members (III: 1); b: the PRRT2 mutation c.971_972insG (p.V325Sfs. 16) was detected in pedigree B to be present in proband its mother (II:1 and I:2), but not in normal pedigree members (I: 1); c: PRRT2 mutation c.959C > T (p.A320V) is a head mutation, which is detected only in proband (II:1) and is not present in parents (I:1 and I: 2); d: detection of the PRRT2 mutation c.1012+5g > a in pedigree D was detected in proband (III:1) and mother (II:2), but not in normal persons (II:1 and II:3) in pedigree. In the figure, the family is shown at the top left, the corresponding generation of sequencing graph is shown at the top right, the proband is indicated by an arrow, and species conservation analysis is shown below.

Fig. 2 is a diagram of clinical features of 284 cases of provers of paroxysmal movement-induced dyskinesia in the present invention, and the percentage of each clinical feature is labeled in a histogram.

Detailed Description

The present invention will now be described in detail with reference to examples and drawings, but the practice of the invention is not limited thereto.

The reagents and starting materials used in the present invention are commercially available or can be prepared according to literature procedures. Experimental procedures without specific conditions noted in the following examples, generally following conventional conditions such as Sambrook et al molecular cloning: the conditions described in the Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), either according to the usual conditions or according to the conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are by weight.

Example detection of four mutant targets of PRRT2 pathogenic Gene

1. Sample collection

284 confirmed peripheral blood specimens of paroxysmal dyskinesia, which were assessed via neurologic clinical assessment and various auxiliary examinations at rekins hospital affiliated with Shanghai university of medical school of transportation, were collected and placed in a refrigerator at 4 ℃. All patients signed informed consent, and the protocol was approved by the ethical committee of the affiliated rekins hospital of the medical college of shanghai transportation university.

2. Total DNA extraction

(1) Transfer 3ml of anticoagulated blood to a 15ml round bottom centrifuge tube, add 9ml of erythrocyte lysate (1 ×), cover the tube cap tightly and reverse it up and down repeatedly, ice-bath for 30 min.

(2) Centrifuging at 3,000rpm for 10min, discarding supernatant, pouring on clean straw paper to suck off more residual liquid, adding 3ml of nuclear fission solution (1X) and 300. mu.l of 10% SDS into leukocyte at the bottom of tube, and blowing and mixing to make the liquid become viscous.

(3) Mu.l proteinase K was added thereto, and the mixture was mixed by inversion and then placed in a 37 ℃ water bath overnight.

(4) To the solution (3), 3ml of Tris-saturated phenol was added, and the mixture was repeatedly shaken and centrifuged at 3,000rpm for 10 min.

(5) The supernatant was aspirated into a new 15ml round-bottom centrifuge tube, 3ml chloroform/isopropanol mixture (24: 1 by volume) was added, shaken vigorously, and centrifuged at 3,000rpm for 10 min.

(6) And (3) sucking the supernatant into a new centrifugal tube, adding 6ml of absolute ethyl alcohol, and reversely mixing until white floccules are separated out, namely the genome DNA.

(7) The white flocculent DNA was transferred to a clean 1.5ml EP tube, centrifuged at 2,000rpm for 5min, the supernatant discarded and the tube bottom DNA pellet was discarded.

(8) The pellet was gently rinsed with 1ml of 75% ethanol, centrifuged at 2,000rpm for 1min, the supernatant was discarded, and the pellet was inverted on clean straw paper.

(9) Repeating the step (8) once, and drying at room temperature for 15 min.

(10) Dissolving with 500 μ l double distilled water at 65 deg.C for 15min, and shaking overnight.

(11) And detecting the DNA concentration of the mother solution by using a multifunctional microplate reader, and diluting the mother solution as required to obtain a working solution. The mother liquor was stored in a refrigerator at-20 deg.C (long term) and the working solution was stored in a refrigerator at 4 deg.C (short term).

3. PCR reaction

PCR reaction solutions were prepared on ice according to the reaction system shown in Table 1.

TABLE 1 PCR reaction System

PCR instrument program: pre-denaturation: 95 ℃ 5min → denaturation: 95 ℃ 30s → annealing: respective annealing temperature 30s → extension: 35-50s at 72 ℃, and 35 cycles of returning to the second step of amplification → 72 ℃ for 10min → 4 ℃ Forever.

The sequences of the primers used in the reaction system are shown in Table 2:

TABLE 2 primer sequences for PRRT2 gene

4. Results

The sequencing results were analyzed by bioinformatics and found to have 4 unreported pathogenic mutations of PRRT2 gene, namely c.1012+5g > a, c.959c > T (p.a320v), c.971_972insG (p.v325sfs 16) and c.761c > T (p.p254l), as detailed in table 3.

TABLE 3 frequency and rating evidence of six disease-causing gene mutations

(Pathogenicity evidence ofsixpathogenic and likelypathogenic variants)

PolyPhen 2 ═ Polymorphism photosetting v 2; 1000g 1000genome Project; ESP6500 ═ NHLBI GO outer Sequencing Project; (ii) an extrame Aggregation Consortium; i not available; d ═ damaging; polyphen-2 scores ranged from 0-1, ≦ 0.2 considered detrimental variation, 0.2-0.85 considered potentially detrimental variation, and >0.85 considered detrimental variation.

a recommends the reference sequence NM-145239 (PRRT 2).

FIG. 1 is a graph showing the detection pedigree of the four mutant targets, and as shown in FIG. 1, PRRT2 mutant c.761C > T (p.P254L) was detected in pedigree A, and was present in proband brother (II:2 and II:1), but was not present in normal pedigree members (III: 1); the PRRT2 mutation c.971_972insG (p.V325Sfs. 16) was detected in pedigree B to be present in proband its mother (II:1 and I:2), but not in normal pedigree members (I: 1); the PRRT2 mutation in pedigree C c.959C > T (p.A320V) is a head mutation, which is detected only in proband (II:1) but not in parents (I:1 and I: 2); detection of the PRRT2 mutation c.1012+5g > a in pedigree D was detected in proband (III:1) and mother (II:2), but not in normal persons (II:1 and II:3) in pedigree. In the figure, the family is shown at the top left, the corresponding generation of sequencing graph is shown at the top right, the proband is indicated by an arrow, and species conservation analysis is shown below.

Fig. 2 is a diagram of 284 cases of paroxysmal motor-induced dyskinesia probands in the present invention.

Sudden movements are symptoms in all patients, with the next highest frequency of symptoms being pre-onset presbycusis (premonition), Dystonia (Dystonia only), and facial involvement (face invasion), as shown in figure 2.

While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention, and the scope of the appended claims is to be accorded the full scope of the invention.

SEQUENCE LISTING

<110> Renjin Hospital affiliated to Shanghai university of transportation medical school

<120> Gene mutation target related to paroxysmal movement induced dyskinesia and application thereof

<130> specification of claims

<160> 8

<170> PatentIn version 3.5

<210> 1

<211> 20

<212> DNA

<213> Artificial sequence

<400> 1

caaatccccg tctcttccag 20

<210> 2

<211> 20

<212> DNA

<213> Artificial sequence

<400> 2

atctccttcg atcccctgtc 20

<210> 3

<211> 20

<212> DNA

<213> Artificial sequence

<400> 3

cccaagccta tctcctcctc 20

<210> 4

<211> 20

<212> DNA

<213> Artificial sequence

<400> 4

ccccattctc ttgcttttcc 20

<210> 5

<211> 19

<212> DNA

<213> Artificial sequence

<400> 5

ccagattccc agcctaccc 19

<210> 6

<211> 20

<212> DNA

<213> Artificial sequence

<400> 6

ggatccatgc agagaggaga 20

<210> 7

<211> 20

<212> DNA

<213> Artificial sequence

<400> 7

ctcccttacc cgccatctat 20

<210> 8

<211> 20

<212> DNA

<213> Artificial sequence

<400> 8

atgcaggagc aggacagaag 20

Claims

1. The application of the reagent for detecting the gene mutation target point in preparing the diagnostic kit of the paroxysmal movement-induced dyskinesia is characterized in that the gene mutation target point is the c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites of PRRT2 gene.

2. The use according to claim 1, wherein the reagent is selected from PCR primers with detection specificity for PRRT2 gene, and the primer sequence is shown as SEQ ID No. 1-SEQ ID No. 8.

3. A diagnostic kit for paroxysmal movement-induced dyskinesia, the kit comprising reagents for detecting the c.1012+5g > a, c.959c > T, c.971_972insG and c.761c > T mutation sites of PRRT2 gene in a biological sample.

4. The diagnostic kit of claim 3, wherein the biological sample is obtained from peripheral blood of the subject.

5. The diagnostic kit of claim 3, wherein the kit comprises: (i) reagents for detecting effective amounts of c.1012+5g > a, c.959C > T, c.971_972insG and c.761C > T mutation sites; (ii) one or more selected from the group consisting of: containers, instructions for use, positive control sequences, negative control sequences, buffers, aids or solvents for cell vortexing or fixation, cell lysis, or nucleic acid purification.

6. The diagnostic kit for paroxysmal movement induced dyskinesia according to claim 5, wherein the kit further comprises instructions for using the kit, wherein the instructions describe how to use the kit for detection, how to use the detection result to judge the development of paroxysmal movement induced dyskinesia, and how to select a treatment regimen.