CN109750101A - Detect gene panel and its application of monogenic inheritance hypertension - Google Patents
Detect gene panel and its application of monogenic inheritance hypertension Download PDFInfo
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Abstract
The invention discloses a kind of gene panel for detecting monogenic inheritance hypertension and its applications.Utilize gene panel of the invention, it can be mutated by targeting high-flux sequence and in conjunction with the potential pathogenic gene that bioinformatic analysis means filter out monogenic inheritance hypertension from magnanimity hereditary information, the positive rate in the mutational site of monogenic inheritance hypertension can be significantly improved；The testing result of the gene panel is combined with the clinical phenotypes of patient, the Etiologic of monogenic inheritance hypertension can effectively be assisted, to improve clinical disease diagnosis rate and implement specific effective therapeutic intervention scheme for particular condition, the early discovery of monogenic inheritance hypertension, the early effect intervened and improve prognosis are realized.Technology of the invention has the remarkable advantages such as fast, the medical diagnosis on disease accuracy height of genetic test speed.
Description
Technical field
The present invention relates to biomedicine technical fields, and in particular to the gene panel of detection monogenic inheritance hypertension
And its application.
Background technique
In secondary hypertension disease, there is one group of disease to meet mendelian inheritance, referred to as monogenic inheritance is high
Blood pressure.Monogenic inheritance hypertension can frequently result in blood pressure raising, electrolyte and the hormonal readiness disorder, medicine of threat to life
Object is resisted, and greatly increases cardiovascular event and early dead risk, particularly with children and Young Adults patient.In clinical work
In work, about 25% refractory hypertension patient suffers from primary aldosteronism, and accounts about 0.1%-1.0% high
Blood pressure patient suffers from pheochromocytoma and Chromaffionoma.
The big feature of the two of monogenic inheritance hypertension are as follows: high genetic and high genetic heterogeneity.It is lost for single-gene
Transmissibility hypertension, from genetics research of generation nineteen thirty based on family, to high-flux sequence in recent years, it has been found that 16
37 pathogenic related genes in seed type monogenic inheritance hypertension.In view of the height heredity of monogenic inheritance hypertension
Degree, genetic test have been recommended in the diagnostic work of a variety of monogenic inheritance Type of Hypertension.
Different from essential hypertension, once really breaking, most secondary hypertensions are can be obtained by etiological treatment
Arrive healing.Thus it is possible to identify monogenic inheritance hypertensive patient from clinic, it is very important, because this is related to
And to specific intervention stratege selection (e.g., glucocorticoid-suppressible aldosteronism need to use hormone therapy,
Gordon syndrome need to use thiazide diuretic to treat).
In recent years, it is sequenced compared to a traditional generation, the development of high-flux sequence (NGS) has greatly improved Meng De
That diagnosis of hereditary disease and the discovery in mutational site, and significantly improve the diagnosis of such disease.But for single-gene
The large-scale genetics research of genetic hypertension pathogenic related gene, either in general population, or in hypertension people
In group, report all there is no at present.
Gene panel is an assortment of genes, using gene panel gene detected than single in genetic test
Site will it is more, than round pcr detect sequence it is long, comparatively, the gene information amount of acquisition wants more.Each base
Because the gene dosage in panel is not of uniform size, people often select different genes to combine according to different testing goals
(panel) related gene in, and there is the decoding capability for these related genes, gene can be just given full play in this way
The effect of panel detection.
In conclusion if the patient's condition in the numerous types of monogenic inheritance hypertension to patient can be efficiently solved
The problem of being made a definite diagnosis will improve clinical disease diagnosis rate and can implement specific effective therapeutic intervention side for particular condition
Case.
Summary of the invention
In order to solve the above technical problems, we have carried out an extensive, polycentric science of heredity in Chinese population
Research.Biological sample such as DNA sample source are related in 1179 clinics of Chinese 19 provinces, 4 municipalities directly under the Central Government, 3 autonomous regions
Suspicious monogenic inheritance hypertensive patient (case crowd) and 1256 healthy population (control) samples.By for by
The monogenic inheritance hypertension relative gene panel of 37 genes composition is sequenced, for example, using Agilent exon
Group capture kit carries out library construction to the sample in experiment, and is sequenced in illumina X Ten platform, described in establishing
The full exon sequencing data storehouse of control crowd (is strong by 1256 inside the Science and Technology Co., Ltd. of Beijing source Nuo Hezhi
The normal healthy controls database that the full sequencing of extron group data of health people are constituted, for filtering pathogenic lower variation) and it is described
Case crowd targeted capture sequencing data storehouse (be 1179 case samples being included in after high-flux sequence, it is resulting prominent
Become the case database that information is constituted, wherein containing large quantities of potential pathogenicity variations), to probe into the high blood of monogenic inheritance
The gene tester of pressure, and combine corresponding Clinical symptoms to carry out the patient's condition and make a definite diagnosis, as early as possible patient be directed to reach
Effective treatment of property.
The present invention one is designed to provide a kind of gene panel for detecting monogenic inheritance hypertension, by 37 lists
The related gene of gene inheritance hypertension forms, and corresponds to Gene Name and is listed below that (relevant information of the gene can join
See online " mankind's Mendelian inheritance " (OMIM, Online Mendelian Inheritance in Man) database
Https: //www.omim.org/):
AIP；ARMC5；CACNA1D；CACNA1H；CUL3；CYP11B1；CYP17A1； CYP21A2；EGLN1；EGLN2；
EPAS1；FH；HSD11B2；KCNJ5；KLHL3； MAX；MEN1；NF1；NOS3；NR3C1；NR3C2；PDE11A；PDE3A；
PDE8B； PRKAR1A；RET；SCNN1B；SCNN1G；SDHA；SDHAF2；SDHB；SDHC；SDHD；TMEM127；VHL；WNK1；
And WNK4.
Another object of the present invention is to provide the base being made of 37 monogenic inheritance hypertension relative genes
Because of application of the panel in the product of preparation detection monogenic inheritance hypertension such as detection device or detection kit.
A purpose of the invention, which also resides in, provides a kind of screening technique of gene mutation, wherein utilizing for test object
Above-described gene panel passes through high-flux sequence and bioinformatic analysis is combined to carry out data processing and gene mutation
Screening.
In some embodiments of the above method of the present invention, data processing and gene in the bioinformatic analysis
The step of screen mutation, is as follows:
A) it was carried out using C language programming software lower machine data (raw data) original to the high-flux sequence of test object
Filter with obtain it is clean read (clean reads), be completely read as Q20 after filtering and be greater than 95%, Q30 being greater than wherein described
85%, and quality data of the G/C content between 40%-60%；
B) above-mentioned clean reading and human genome reference sequences are subjected to sequence alignment to obtain the format of sequence alignment
File, wherein the target comparison rate of the sequence alignment is 95% or more, average sequencing depth > 100 ×, mean coverage >
98.9%；
C) site annotation is carried out by functional annotation software and screens potential disease cause mutation site.
In some embodiments of the above method of the present invention, original lower machine data are filtered in step a) original
Then are as follows: (1) filter the pairs of reading (paired reads) containing connector；(2) filtering N content is more than the pairs of of reading 10%
It reads；(3) sequence that low quality base (Qphred≤5) content in single reading is more than 50% is filtered.
In some embodiments of the above method of the present invention, the target comparison rate of sequence alignment described in step b)
(mapping rate) 95% or more, average sequencing depth > 100 ×, mean coverage > 98.9%, and the sequence alignment
As a result formatted file is bam file.
In some embodiments of the above method of the present invention, carried out in step c) by functional annotation software ANNOVAR
Site annotation and screening, detailed process are as follows:
(1) mutation of frequency of occurrence >=1 time in the full exon sequencing data storehouse of normal healthy controls crowd's sample is filtered out；
(2) mutation of frequency of occurrence >=100 time in the targeted capture sequencing data storehouse of case crowd's sample is filtered out；
(3) mutation recall rate is filtered out to be higher than in normal healthy controls crowd's sample database in case crowd's sample database
In mutational site；
(4) sequencing coverage DP4 < 4, and the mutational site of DP4/DP < 0.1 are filtered out；
(5) site except the panel of 37 genes of monogenic inheritance hypertension and non-exon region are filtered out
Site；
(6) it filters out and organizes integrated database (ExAC, Exome in international human genome project database, human exonic
Aggregation Consortium), in normal healthy controls crowd's sample database minimum gene frequency MAF > 0.01 position
Point；
(7) retain the site being incorporated in HGMD database；
(8) retain a variety of bioinformatics softwares to be predicted as causing a disease, may cause a disease, the site that clinical meaning is unknown；
(9) retain the site of > 10 points of CADD scoring.
One aspect of the invention also provides said gene screen mutation method in auxiliary diagnosis monogenic inheritance hypertension
Disease because device in application.
Another aspect of the invention also provides a kind of device of auxiliary diagnosis monogenic inheritance hypertensive patient's cause of disease,
Comprising: in the gene panel 37 genes targeted capture probe, the primer pair of 37 genes is expanded, to targeting
The filtering module that the original lower machine data of high-flux sequence are filtered, by the target sequence of object to be measured and reference sequences into
The sequence alignment module that row compares, and the sequence alignment result is annotated and screens potential pathogenic mutation gene
Genescreen module.
The potential disease cause mutation site that the present invention is filtered out can assist high blood in conjunction with the clinical phenotypes of patient
Press the Etiologic of patient.
Beneficial effect
By high-flux sequence (NGS) technology, the gene of detection monogenic inheritance hypertension of the invention is utilized
Panel can disposably carry out multiple monogenic inheritance hypertension relative genes that screening is sequenced without bias, then utilize biology
Bioinformatics analysis means from magnanimity hereditary information by annotation, filtering and etc. filter out potential disease cause mutation, Neng Gouxian
Write the positive rate for improving the mutational site of monogenic inheritance hypertension；By the testing result of the gene panel and trouble
The clinical phenotypes of person combine, and can effectively assist the Etiologic of monogenic inheritance hypertensive patient, face to improve
Bed medical diagnosis on disease rate simultaneously implements specific effective therapeutic intervention scheme for particular condition, realizes monogenic inheritance hypertension
Early discovery, the early effect intervened and improve prognosis.Technology of the invention fast, the medical diagnosis on disease accuracy with genetic test speed
High remarkable advantage.
Detailed description of the invention
Fig. 1 is the both adrenal glands enhanced CT figure of women multiple endocrine neoplasia (MEN) 2B patient Ren: on right kidney
Visible bulk capsule reality density lump shadow in gland area, sharpness of border, 72 × 55mm, Density inhomogeneity in stove are thought of as thermophilic chromium
Cytoma；Left adrenal gland thickening, is thought of as adrenal hyperplasia.
Fig. 2 is to be related to MEN 2B patient Ren (I2) and husband (I1) and its 4 name woman's pedigree charts, and wherein I is indicated
Parental generation, II indicate filial generation, and square indicates male, and circle indicates that women, solid black graphical representation have appearance abnormal appearance
Disease affected individual, hollow graphical representation do not correspond to the healthy family member of appearance abnormal appearance；I1 is 41 years old, and I2 is 37 years old
(arrow is directed toward propositus Ren), II1 are 13 years old, and II 2 is 11 years old；II 3 is 10 years old, and II 4 is 9 years old.
Fig. 3-6 is MEN 2B propositus Ren (I generation) corresponding with Fig. 2 pedigree chart and its disease of children's (II generation)
Related appearance external appearance characteristic picture, wherein Fig. 3 be I2 (propositus Ren), Fig. 4 be II2 (affected individual), Fig. 5 be II3 (by
Tired person), Fig. 6 is II4 (affected individual).
Fig. 7 is II1 (non-affected individual).
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, implementing
The conventional means that technological means used in example is well known to those skilled in the art.
Embodiment
The diagnosis of family 1,1 multiple endocrine neoplasia 2B type of embodiment and Analysis of clinical features
2 type of multiple endocrine neoplasia (MEN 2) is a kind of neoplastic syndrome of autosomal dominant inheritance.According to
Its clinical manifestation, pathological characteristics and genetic base difference, are divided into 3 kinds of hypotypes, respectively MEN 2A, MEN 2B, MTC.Its
Although middle MEN 2B incidence is only 2.5/10 ten thousand, because it has the characteristics that age of onset is early, invasion is high, case fatality rate is high,
Therefore early diagnosis, early intervention it is particularly critical.The present embodiment is to other 5 familys in 1 MEN 2B propositus and its family
The Clinical symptoms of member is analyzed.
1, case basic condition:
Patient Ren, women, 37 years old, in August, 2012 was because " palpitaition is fainted April repeatedly, is aggravated with nausea and vomiting 20
It " it goes to a doctor in the second affiliated hospital, University Of Nanchang, it is admitted to hospital with " palpitaition is to be checked ".Zeng Yin " thyroid cancer " row " left side in 2009
Subtotal thyroidectomy ", readme is without hypertension history.It is married, 2 sub 2 female are given birth to, wherein 2 sub 1 female's appearance appearances have exception.
It is admitted to hospital tentative diagnosis " palpitaition syncope is to be checked ".
2, physical examination:
Be admitted to hospital blood pressure 120/86mmHg, and 112 beats/min of pulse.Lip thickens, visible number at back, lingual margin mucous membrane and frenulum
A thin tumor sample object of kick shape not of uniform size.The visible operative incision scar of thyroid gland.
3, auxiliary examination:
Patient improves the double kidney color ultrasounds of liver and gallbladder pancreas spleen after being admitted to hospital: margo interior hepatis and right upper pole of kidney mixing rise agglomerate, 62 ×
57mm, however not excluded that adrenal gland source tumour.After improve both adrenal glands enhanced CT: visible bulk capsule reality in right adrenal gland area
Density lump shadow, sharpness of border, 72 × 55mm, Density inhomogeneity in stove are thought of as pheochromocytoma；Left adrenal gland thickening, is examined
Consider is adrenal hyperplasia (Fig. 1).Have a blood test cortisol rhythm, parathyroid hormone, feritin, aldosterone level is normal.Repeatedly look into life
Change blood glucose slightly to increase, glycosylated hemoglobin slightly increases.Urine proteins (2+), vanilmandelic acid (VMA) are normal.
4, clinical diagnosis:
Because patient is there are paroxysmal palpitaition pre-syncopal symptoms, human health screening finds the thin tumor sample protrusion tissue in lip portion, is thought of as
Mucous membrane neuroma, there is right side Adrenal Pheochromocytoma in adrenal CT prompt, and previously have thyroid cancer, left thyroid gland to cut
Except history, therefore consider that patient is diagnosed as multiple endocrine neoplasia MEN 2B.
5, the Analysis of clinical features of family member
In view of MEN 2B is monogenic inheritance neoplastic syndrome, there is high genetic tendency, we are to propositus Ren
The family member of certain (patient) has carried out Analysis of clinical features.As a result, it has been found that patient husband is that related appearance appearance is normally good for
Health people, but there is the appearance appearance of 3 people to have exception, and the thin tumor sample protrusion in visible lip portion in its 2 sub 2 female, there is 1 people for related phase
The normal Healthy People of looks appearance (pedigree chart is as shown in Fig. 2, propositus and children's face off-note are shown in Fig. 3-6)；In short, packet
4 afflicted individuals (I2, II2, II3 and II4) for including propositus have unusual facies, show as lip and thicken, back,
Visible multiple sizes of thin tumor sample shape protrusion at lingual margin mucous membrane and frenulum considers mucous membrane of mouth neuroma, belongs to multiple
Property endocrine neoplasia MEN 2B；And the visible left postoperative scar of thyroidectomy of propositus (I 2) neck；And non-afflicted individual (II
1) without unusual facies.
Embodiment 2 compares the foundation of crowd's full exon sequencing and case crowd targeted capture sequencing data storehouse
1, biological specimen collection
DNA sample source is related to suspicious in 1179 clinics of Chinese 19 provinces, 4 municipalities directly under the Central Government, 3 autonomous regions
Monogenic inheritance hypertensive patient (case crowd) and 1256 healthy population (controls.
2, the foundation of database
Using the monogenic inheritance hypertension relative gene panel cited hereinabove being made of 37 genes to this
A little samples are sequenced, and carry out library structure to the sample in experiment using Agilent exon group capture kit in the present invention
It builds, and is sequenced in illumina X Ten platform, it is entirely outer to establish case crowd's targeted capture sequencing data storehouse and control crowd
Aobvious son sequencing.
Main purpose of the building for the cDNA library of high-flux sequence is all to connect and want at the both ends of DNA fragmentation
Connector.CDNA library building process specifically includes that 1, fragmentation DNA；2, end-filling；3, in 3 ' ends plus polyA tail；4,
It is connected with connector；5, PCR amplification.The cDNA library of high quality need containing respectively with P5 and P7 connector on the surface FlowCell
Identical structure.
3, result
The case crowd's targeted capture sequencing data storehouse established is that 1179 case samples being included in are measured by high pass
After sequence, the case database that resulting abrupt information is constituted, wherein containing large quantities of potential pathogenicity variations.
The full exon sequencing data storehouse of the control crowd established is inside the Science and Technology Co., Ltd. of Beijing source Nuo Hezhi
The normal healthy controls database being made of the full sequencing of extron group data of 1256 Healthy Peoples, it is pathogenic lower for filtering
Variation.
Embodiment 3 detects multiple endocrine neoplasia 2B type man using monogenic inheritance shr gene detection panel
It is 1
1, sample collection and genetic test:
In view of RET Oncogene Mutation it is the pathogenesis basis of MEN 2, and is related to RET proto-oncogene Met918Thr mutation
Multiple endocrine neoplasia MEN 2B family report, we plan to this 6 family members carry out targeting sequencing gene inspection
It surveys, analyzes the gene mutation situation of the family.We have collected patient Mr. and Mrs and 2 sub 2 female, the buccal swab DNA sample of totally 6 people
This, for 37 monogenic inheritance hypertension pathogenic related genes including MEN 2B Disease-causing gene RET proto-oncogene
Exon region carries out targeting sequencing (described 37 using monogenic inheritance shr gene detection panel of the invention
Monogenic inheritance hypertension pathogenic related gene is for example cited hereinabove).
The present invention captures kit to the sample progress library construction in experiment using Agilent exon group, and in
The sequencing of illumina X Ten platform.Basic process are as follows: after the frag-ment libraries for obtaining 500bp or so, it is sequenced for upper machine,
To obtain original lower machine data (raw data).
Then, data processing is carried out by bioinformatic analysis and gene mutation is screened, the specific steps are as follows:
A) internal C language programming software is used, it is original to the high-flux sequence of 6 test objects of above-mentioned family member
Lower machine data are filtered the clean reading (clean reads) of acquisition；Filter principle are as follows: (1) filter the pairs of reading containing connector
Take (paired reads)；(2) filtering N content is more than the pairs of reading for reading 10%；(3) low-quality in the reading of filtering single
Amount base (Qphred≤5) content is more than 50% sequence；Reading Q20 after filtering is greater than 95%, Q30 and is greater than 85%,
And the data of high quality are used for subsequent analysis in suitable range by G/C content；
B) above-mentioned clean reading is compared with human genome reference sequences using bwa software, to obtain sequence alignment
Bam file；Wherein the target comparison rate of the sequence alignment is 95% or more, and average sequencing depth > 100 ×, mean coverage
> 98.9%；
C) site annotation is carried out by functional annotation software ANNOVAR and screens potential disease cause mutation site, had
Body process are as follows:
(1) frequency of occurrence >=1 in normal healthy controls crowd (1256 healthy full exon sequencings of sample) database is filtered out
Secondary mutation；
(2) frequency of occurrence >=100 time in case crowd (1179 clinical samples targeted capture sequencings) database are filtered out
Mutation；
(3) mutation recall rate is filtered out in normal healthy controls crowd's sample database higher than case crowd sample database
Mutational site；
(4) sequencing coverage (DP4) < 4, and the mutational site of DP4/DP < 0.1 are filtered out；
(5) site except the panel of 37 genes of monogenic inheritance hypertension and non-exon region are filtered out
Site；
(6) it filters out in international human genome project database, ExAC database, 1256 normal healthy controls crowd's samples
In database, the site of minimum gene frequency MAF > 0.01；
(7) retain the site being incorporated in HGMD database；
(8) retain a variety of bioinformatics softwares to be predicted as causing a disease, may cause a disease, the site that clinical meaning is unknown；
(9) retain the site of > 10 points of CADD scoring.
2, testing result
Testing result shows that propositus I2 detects the c.2753 position nucleosides of the 16th exon of MEN 2B Disease-causing gene RET
Sour heterozygosity c.T2753C mutation, causes the 918th amino acids that p.M918T mutation occurs, corresponding methionine is caused to be mutated
For threonine (Met918Thr).And then same other 5 family members for receiving sequencing are analyzed, wherein appearance has exception for discovery
There is this mutation in sub 1 female in the 2 of performance, i.e. II2, II3 and II4.And propositus husband I1 and 1 daughter II1 are not detected this and dash forward
Become (targeting sequencing result is shown in following table 1).Therefore consider, 3 appearances for detecting mutation have the face of abnormal children abnormal prominent
It rises and is showed for oral mucosa neuroma, belong to multiple endocrine neoplasia MEN 2B.
Table 1: propositus and its family member target sequencing result
Three, it treats
Patient Ren is diagnosed as MEN 2B, and is interrupted breaking-out palpitaition, syncope, influences quality of life, it is proposed that be transferred to uropoiesis
The operation excision of surgery row pheochromocytoma.But patient is transferred to Urology Surgery and waits during operation because operation risk is by refusing
Exhausted operative treatment is then left hospital.Suggest that patient periodically answers in institute's layman's specific treatment, and periodically in Cardiological follow-up after discharge
The imageological examinations such as adrenal CT, color ultrasound are looked into, periodic review urinates VMA, blood cortisol, thyroid hormone, parathyroid hormone etc.
Index is vigilant the generation of other endocrine system carcinomas.
In short, panel is detected by using monogenic inheritance shr gene of the invention, in conjunction with other every phases
The incidence of the multiple endocrine neoplasia 2B type family generation of neutrons of patient Ren has been made a definite diagnosis in the clinical examination of pass, is real
It applies specific effective therapeutic intervention scheme and provides exact foundation.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to claimed model without departing from theon the basis of the spirit of the present invention
It encloses.
Claims (10)
1. a kind of gene panel for detecting monogenic inheritance hypertension, which is characterized in that by 37 high blood of monogenic inheritance
The related gene of pressure forms, and the title of the gene is listed below:
AIP；ARMC5；CACNA1D；CACNA1H；CUL3；CYP11B1；CYP17A1；CYP21A2；EGLN1；EGLN2；EPAS1；
FH；HSD11B2；KCNJ5；KLHL3；MAX；MEN1；NF1；NOS3；NR3C1；NR3C2；PDE11A；PDE3A；PDE8B；
PRKAR1A；RET；SCNN1B；SCNN1G；SDHA；SDHAF2；SDHB；SDHC；SDHD；TMEM127；VHL；WNK1；And WNK4.
2. application of the gene panel described in claim 1 in the product of preparation detection monogenic inheritance hypertension.
3. application as claimed in claim 2, which is characterized in that the product is detection device or detection kit.
4. a kind of screening technique of gene mutation, which is characterized in that utilize gene described in claim 1 for test object
Panel passes through high-flux sequence and bioinformatic analysis is combined to carry out data processing and gene mutation screening.
5. method as claimed in claim 4, which is characterized in that the data processing and gene mutation of the bioinformatic analysis
The step of screening, is as follows:
A) it is filtered using the original lower machine data of high-flux sequence of the C language programming software to test object to obtain clean reading
It takes, is completely read as Q20 after filtering and is greater than 95%, Q30 being greater than 85% wherein described, and G/C content is between 40%-60%
Quality data；
B) above-mentioned clean reading is obtained to the formatted file of sequence alignment with human genome reference sequences progress sequence alignment,
Wherein the target comparison rate of the sequence alignment is 95% or more, and average sequencing depth > 100 ×, mean coverage > 98.9%；
C) site annotation is carried out by functional annotation software and screens potential disease cause mutation site.
6. method as claimed in claim 5, which is characterized in that the principle being filtered in step a) to original lower machine data
Are as follows: (1) filter the pairs of reading containing connector；(2) filtering N content is more than the pairs of reading for reading 10%；(3) filtering single is read
Take middle Qphred≤5 low quality base contents be more than 50% sequence.
7. method as claimed in claim 5, which is characterized in that the target comparison rate of sequence alignment described in step b) is 95%
More than, average sequencing depth > 100 ×, mean coverage > 98.9%, and the formatted file of the sequence alignment result is bam text
Part.
8. method as claimed in claim 5, which is characterized in that carry out position by functional annotation software ANNOVAR in step c)
Point annotation and screening, detailed process are as follows:
(1) mutation of frequency of occurrence >=1 time in the full exon sequencing data storehouse of normal healthy controls crowd's sample is filtered out；
(2) mutation of frequency of occurrence >=100 time in the targeted capture sequencing data storehouse of case crowd's sample is filtered out；
(3) it filters out mutation recall rate and is higher than dashing forward in case crowd's sample database in normal healthy controls crowd's sample database
Displacement point；
(4) sequencing coverage DP4 < 4, and the mutational site of DP4/DP < 0.1 are filtered out；
(5) site except the panel of 37 genes of monogenic inheritance hypertension and the position of non-exon region are filtered out
Point；
(6) it filters out minimum in international human genome project database, ExAC database, normal healthy controls crowd's sample database
The site of gene frequency MAF > 0.01；
(7) retain the site being incorporated in HGMD database；
(8) retain a variety of bioinformatics softwares to be predicted as causing a disease, may cause a disease, the site that clinical meaning is unknown；
(9) retain the site of > 10 points of CADD scoring.
9. gene mutation screening technique as claimed in claim 4 is in the dress of auxiliary diagnosis monogenic inheritance hypertensive patient's cause of disease
Application in setting.
10. the device of auxiliary diagnosis monogenic inheritance hypertensive patient's cause of disease, which is characterized in that including described in claim 1
The targeted capture probe of 37 genes in gene panel expands the primer pair of 37 genes, to targeting high-flux sequence
The filtering module that original lower machine data are filtered, the sequence ratio that the target sequence of object to be measured is compared with reference sequences
To module, and the sequence alignment result is annotated to and is screened the genescreen module of potential pathogenic mutation gene.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111653316A (en) * | 2020-05-27 | 2020-09-11 | 上海寻因生物科技有限公司 | Visualization analysis method, system and storage medium based on next generation sequencing |
| CN112608994A (en) * | 2020-12-31 | 2021-04-06 | 南昌瑞因康生物科技有限公司 | Detection method, device and storage medium for hypertrophic cardiomyopathy and related genes |
| CN113684220A (en) * | 2021-09-06 | 2021-11-23 | 百世诺（北京）医疗科技有限公司 | Single gene hypertension gene detection kit based on mutated VHL gene |
| CN113957141A (en) * | 2021-12-01 | 2022-01-21 | 长沙艾迪康医学检验实验室有限公司 | Oligonucleotide for detecting hypertension-related gene SCNN1B mutation and application thereof |
| KR20220022313A (en) * | 2020-08-18 | 2022-02-25 | 주식회사 테라젠바이오 | Method for providing information for hypertension and kits using the same |
| CN114381514A (en) * | 2022-01-27 | 2022-04-22 | 圣湘生物科技股份有限公司 | Gene detection panel for detecting adrenal hypertension and application thereof |
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| CN111653316A (en) * | 2020-05-27 | 2020-09-11 | 上海寻因生物科技有限公司 | Visualization analysis method, system and storage medium based on next generation sequencing |
| KR20220022313A (en) * | 2020-08-18 | 2022-02-25 | 주식회사 테라젠바이오 | Method for providing information for hypertension and kits using the same |
| KR102565803B1 (en) | 2020-08-18 | 2023-08-10 | 주식회사 테라젠헬스 | Method for providing information for hypertension and kits using the same |
| CN112608994A (en) * | 2020-12-31 | 2021-04-06 | 南昌瑞因康生物科技有限公司 | Detection method, device and storage medium for hypertrophic cardiomyopathy and related genes |
| CN113684220A (en) * | 2021-09-06 | 2021-11-23 | 百世诺（北京）医疗科技有限公司 | Single gene hypertension gene detection kit based on mutated VHL gene |
| CN113957141A (en) * | 2021-12-01 | 2022-01-21 | 长沙艾迪康医学检验实验室有限公司 | Oligonucleotide for detecting hypertension-related gene SCNN1B mutation and application thereof |
| CN114381514A (en) * | 2022-01-27 | 2022-04-22 | 圣湘生物科技股份有限公司 | Gene detection panel for detecting adrenal hypertension and application thereof |
| CN114381514B (en) * | 2022-01-27 | 2024-05-07 | 圣湘生物科技股份有限公司 | Gene detection panel for detecting adrenal hypertension and application thereof |
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