CN110452980A - A kind of mitochondrial encephalomyopathy diagnostic kit and application - Google Patents
A kind of mitochondrial encephalomyopathy diagnostic kit and application Download PDFInfo
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- CN110452980A CN110452980A CN201910897061.5A CN201910897061A CN110452980A CN 110452980 A CN110452980 A CN 110452980A CN 201910897061 A CN201910897061 A CN 201910897061A CN 110452980 A CN110452980 A CN 110452980A
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
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Abstract
The invention discloses a kind of mitochondrial encephalomyopathy diagnostic kit and applications.The mitochondrial encephalomyopathy diagnostic kit comprising ECHS1 gene order detection reagent.The application of ECHS1 gene order detection reagent is applied to the preparation of mitochondrial encephalomyopathy diagnostic reagent.The present invention passes through the incidence relation between clear ECHS1 gene and mitochondrial encephalomyopathy, the especially rare mutation c.414+5G > A of ECHS1 gene intron 3 leads to mitochondrial encephalomyopathy, provide the application of a kind of mitochondrial encephalomyopathy diagnostic kit and ECHS1 gene order detection reagent, for mitochondrial encephalomyopathy diagnosis and make a definite diagnosis, provide important science of heredity foundation.
Description
Technical field
The invention belongs to medical diagnostic fields, more particularly, to a kind of mitochondrial encephalomyopathy diagnostic kit and application.
Background technique
Related disease of mitochondria damage mainly includes various types of clinical phenotypes mainly as caused by heredity.
Most of cases have brain exception, but seldom clinical case only individually shows as brain exception.In addition, to patient, especially
It is very difficult that the related disease of mitochondria damage of baby, which makes and clarifying a diagnosis,.The various of mitochondrial protein are encoded in karyogene
Genetic mutation may result in mitochondria dysfunction, be the core reasons for causing related disease of mitochondria damage.Understand line grain
Relationship between the clinical manifestation of bulk damage related disease and gene genetic mutation has important meaning to establishing to diagnose and prevent mistaken diagnosis
Justice.
Mitochondrial encephalomyopathy (ME) is caused by one group of rare structure of mitochondria and (or) dysfunction with brain and flesh
Multisystem disease based on meat involvement.Its muscle damage is mainly shown as that skeletal muscle is not resistant to fatigue, nervous system master extremely
It is presented with ballet's disease, stroke, epilepsy recurrent exerbation, myoclonia, migraine, incoordination, disturbance of intelligence and view mind
Through lesion etc., other systems performance can have heart block, cardiomyopathy, diabetes, renal insufficiency, intestinal pseudo obstruction and body
Material is short and small etc..From at present to the research of this disease from the point of view of, think that this disease is that harmful gene and line grain are carried due to patient more
Body function is abnormal, and thus leads to diverse clinical manifestations.It there is no the associated gene of specific mitochondrial encephalomyopathy, at present line grain
Body genetic test result can not make a definite diagnosis mitochondrial encephalomyopathy.
Summary of the invention
Aiming at the above defects or improvement requirements of the prior art, the present invention provides a kind of mitochondria cerebral disease detection reagents
Box and application, its object is to use due to specifying exact incidence relation between ECHS1 gene and mitochondrial encephalomyopathy
ECHS1 gene detection reagent is used for the preparation of mitochondrial encephalomyopathy diagnostic reagent, thus solves current mitochondrial encephalomyopathy and lacks
The technical issues of effectively making a definite diagnosis means.
To achieve the above object, according to one aspect of the present invention, a kind of mitochondrial encephalomyopathy diagnostic kit is provided,
It includes ECHS1 gene order detection reagent.
Preferably, the mitochondrial encephalomyopathy diagnostic kit comprising the gene order of ECHS1 gene intron 3 is examined
Test agent.
Preferably, the mitochondrial encephalomyopathy diagnostic kit comprising the rare mutation of ECHS1 gene intron 3 is c.414
+ 5G > A detection reagent.
Preferably, the mitochondrial encephalomyopathy diagnostic kit, the ECHS1 gene order detection reagent include
ECHS1 gene amplification reagent.
Preferably, the mitochondrial encephalomyopathy diagnostic kit, the ECHS1 gene amplification reagent include:
Forward primer: 5 '-AGCTACACCTGGAGCCACTG-3 '；And:
Reverse primer: 5 '-CCATGGACACGAATCACAAG-3 '.
Preferably, the mitochondrial encephalomyopathy diagnostic kit comprising ECHS1 gene expression detection reagent.
Preferably, the mitochondrial encephalomyopathy diagnostic kit, the ECHS1 gene expression detection reagent include
ECHS1 gene RT-PCR primer:
Forward primer: 5 '-TCTGAGTCACCTGGACAACC-3 '；And:
Reverse primer: 5 '-ATCTCAGTGGTATTTGTCAGC-3 '.
Other side according to the invention provides a kind of application of ECHS1 gene order detection reagent, is applied to
The preparation of mitochondrial encephalomyopathy diagnostic reagent.
Preferably, the application is applied to the abnormal caused mitochondrial encephalomyopathy diagnosis examination of ECHS1 gene intron 3
The preparation of agent.
Preferably, the application is applied to line caused by the rare mutation c.414+5G > A of ECHS1 gene non-coding region
The preparation of plastochondria brain myopathy diagnostic reagent.
In general, through the invention it is contemplated above technical scheme is compared with the prior art, can obtain down and show
Beneficial effect:
The present invention is included by the incidence relation between clear ECHS1 gene and mitochondrial encephalomyopathy, especially ECHS1 gene
3 rare mutation c.414+5G > A of son lead to mitochondrial encephalomyopathy, provide a kind of mitochondrial encephalomyopathy diagnostic kit and
The application of ECHS1 gene order detection reagent is the confirmation of mitochondrial encephalomyopathy, provides important science of heredity foundation.
Detailed description of the invention
Fig. 1 is the MRI testing result of infant of the embodiment of the present invention；
Head B-sonography bilateral basal section thalamus area and midbrain brain symmetry abnormal signal as the result is shown, bilateral frontal temporal part spider
Nethike embrane cavity of resorption is broadening, bilateral volume top temporo occipital lobe cortex, brain stem, basal ganglia region abnormal signal (A-F).
Fig. 2 is the MRS testing result of infant of the embodiment of the present invention；
Ch/NAA value is 3.47 to MRS as the result is shown, compared with normal value, there is reduction.
Fig. 3 is that infant of embodiment of the present invention ECHS gene sequencing result and c.414+5G > A mutation bioinformatics are pre-
It surveys；
Two generation sequencing results show that c.414+5G > A mutation (A) occurs in the introne 3 of infant ECHS1 gene.Using life
Object bioinformatics analysis, c.414+5G > A mutation may result in ECHS1 gene cDNA and the missing (B) of 39bp occurs as the result is shown.
Fig. 4 is that minigene of embodiment of the present invention analysis analyzes c.414+5G > A mutation function；
C.414+5G > A mutation function is observed using minigene.C.414+5G > A as the result is shown of gel electrophoresis experiment
Mutation will cause ECHS1 gene cDNA and the missing (A) of 39bp occurs.B is that c.414+5G > A mutation causes ECHS1 gene cDNA to go out
The ideograph of existing 39bp missing.Using Sanger sequence verification c.414+5G > A mutation cause ECHS1 gene cDNA 39bp occur
Missing (C).
Fig. 5 be Bioinformatics Prediction of the embodiment of the present invention c.414+5G > A be mutated caused by ECHS1 protein structure become
Change；
Bioinformatics Prediction the results show that c.414+5G > A mutation will lead to 13, the enzyme activity region of ECHS1 albumen
The missing of Core amino acids.
Fig. 6 be the embodiment of the present invention carry ECHS1 gene c.414+5G > infant of A mutation, in fibroblast
The activity change of ECHS1 (2- enoyl-CoA hydratase).
Using spectrophotometry, in the infant fibroblast, shadow of the c.414+5G > A mutation to ECHS1 enzyme activity
It rings.The results show that carry ECHS1 gene c.414+5G > infant of A mutation, ECHS1 (2- enoyl CoA in fibroblast
Hydrase) enzyme activity significant decrease.
Specific embodiment
In order to make the objectives, technical solutions, and advantages of the present invention clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.As long as in addition, technical characteristic involved in the various embodiments of the present invention described below each other it
Between do not constitute conflict and can be combined with each other.
ECHS1 gene is located at chromosome 10q26.2-q26.3.The short chain propylene oxygen ethyl of ECHS1 gene code mitochondrial is auxiliary
Enzyme A hydrase (short chain enoyl-CoA hydratase, sceh or echs1), is positioned at mitochondrial matrix, in many metabolic pathways
The aquation of middle catalysis enoyl CoA, including short chain fatty acids beta oxidation and branch.Amino acid catabolic, cyclophorase
It is catalyzed unsaturated fatty acid, simultaneously participates in the branched-aminos acid metabolic such as eucaryote valine, leucine, isoleucine.ECHS1
It is widely studied in model organism due to its special role in mitochondria oxidative function, but ECHS1 and baby are strong before this
Relationship between health and disease is unclear.Inventor has found a kind of new ECHS1 mutation, and provides enough experiment cards
According to showing that the mutation causes mitochondrial encephalomyopathy.
The present invention provides a kind of mitochondrial encephalomyopathy diagnostic kits, comprising: DNA extracts reagent, ECHS1 gene order
Detection reagent and ECHS1 gene expression detection reagent；
The DNA extracts reagent, is complete genome DNA extraction purification reagent, the preferably full base of the peripheral blood containing EDTA
Because group DNA extracts reagent.
The ECHS1 gene order detection reagent, the gene order detection reagent including ECHS1 gene intron 3, including
ECHS1 gene PCR amplifing reagent.The full-length genome that the template that the ECHS1 gene PCR amplifing reagent uses is extracted for peripheral blood
DNA, the primer of use are as follows:
Forward primer: 5 '-AGCTACACCTGGAGCCACTG-3 '；And:
Reverse primer: 5 '-CCATGGACACGAATCACAAG-3 '.
The ECHS1 gene expression detection reagent, including ECHS1 gene expression detection reagent, preferably ECHS1 gene
RT-PCR detection reagent, the primer used are as follows:
Forward primer: 5 '-TCTGAGTCACCTGGACAACC-3 '；And:
Reverse primer: 5 '-ATCTCAGTGGTATTTGTCAGC-3 '.
The present invention specifies the dysfunction of ECHS1 gene, causes mitochondrial encephalomyopathy, therefore ECHS1 gene order is examined
Test agent can apply to the preparation of mitochondrial encephalomyopathy diagnostic reagent；Especially, in ECHS1 gene non-coding region introne 3
The preparation of mitochondrial encephalomyopathy diagnostic reagent caused by rare rare mutation c.414+5G > A of mutation.
The following are embodiments:
Children's hospital, Wuhan City accepts an example infant for medical treatment, is boy baby, and parent's health produces, birth weight 3 for pregnant 40 weeks,
500g, the long 51cm of body.The infant is that the first case, brother have no symptom in family.Its parent is all Chinese, non-close relative's knot
It closes.When the infant is born, sample symptom is embraced in discovery, 5-6 times daily, and night morbidity is more frequent.
When the infant is one month big, because pneumonia of newborn, icterus neonatorum, left uronephrosis are hospitalized.Shi Faxian 8 months big
Its psychomotor development delay.Can't raise one's head at eight months, at 13 months can not sitting alone, when two one full year of life is beyond expression
If one significant.After one one full year of life, spasm, which shows, becomes especially prominent.
The infant is checked, electromyogram and repetition electroencephalogram are normal.Laboratory checks that display lactic acid increases
11.41mmol/L (normal range (NR) 0.5-2.22mmol/L), 201 μm of ol/L of blood ammonia (18-72 μm of ol/L of normal range (NR)).In blood
It is without exception that fatty acyl carnitine analyzes result.Urinary organic acid is analysis shows that the excretion of 3- hydroxybutyl carnitine dramatically increases.Urinalysis
It has been shown that, ketone 3+, bil1+, pro1+.Excrement, which is occulted blood, checks the positive, and prompting patient, there are intestinal mucosa injuries.T3, T4 and TSH are horizontal
In the normal range.Liver and kidney function, myocardium enzyme, electrolyte level are normal.We judge infant for " mitochondria brain flesh
Disease ", and trimestral cocktail therapy has been carried out to it.After treatment, symptom is obviously improved, and embraces the reduction of sample seizure frequency.
Brain magnetic resonance imaging shows, bilateral forehead occipital cortex, brain stem and Basal ganglia there are dispersivity, length and it is symmetrical
T1 and T2 signal distributions.DWI shows high RST.Bilateral telocoele is slightly larger, and brain ditch is slightly wider.Posterior fossa cerebellum shows no obvious abnormalities letter
Number, centerline construction has no displacement (Fig. 1).NAA declines MRS as the result is shown, and CH/NAA declines 3.47 (Fig. 2).
DNA is extracted:
DNA extracts reagent: complete genome DNA is extracted and purified from infant peripheral blood.Peripheral blood DNA mentions in the present embodiment
The peripheral blood genome for taking use " Axygen Scientific " company to produce extracts and purification kit.It is firstly added 500 μ L
Ultrapure water sterilize into 200 μ L peripheral bloods, splitting erythrocyte.Then it at 4 DEG C, is centrifuged 15 minutes, discards supernatant, it is heavy to retain
It forms sediment, is precipitated as leucocyte.The Proteinase K of Roche company production is added, cracks leucocyte.Isopropanol is added, genome is precipitated
DNA is eventually adding 75% ethyl alcohol rinsing.
ECHS1 gene magnification:
Amplimer:
Forward primer: 5 '-AGCTACACCTGGAGCCACTG-3 ' (SEQ No.1)；And:
Reverse primer: 5 '-CCATGGACACGAATCACAAG-3 ' (SEQ No.2).
It uses complete genome DNA for template, PCR amplification ECHS1 gene extron 2, exon 3, exon 4 and includes
Sub 2, introne 3 sequence, with ABI PRISM 3500xL Genetic Analyser (Applied Biosystems, USA) direct Sequencing
Analyze PCR product.ECHS1 gene gene number in NCBI omim database is 616277, and number is in ENSEMBL
The gene order of ENSG00000210049.
XhoI is used to above-mentioned PCR product, BamHI restriction enzyme carries out digestion to generate cohesive end, then by the base
Because PCR product is cloned into pSPL3 carrier, and whether validating DNA segment is correctly inserted into.By wild type carrier and inserted with c.414+
The carrier of 5G > A mutation ECHS1 gene is transferred to African green monkey kidney COS7 cell line and carries out montage analysis, after transfection in 48 hours,
It extracts total serum IgE and carries out RT-PCR.
ECHS1 gene expression RT-PCR detection:
RNA is extracted and the specific method of RT-PCR is shown in " Qing brick tea (QBT) aqueous extract
protects monosodium glutamate-induced obese mice against metabolic syndrome
and involves up-regulation Transcription Factor Nuclear Factor-Erythroid 2-
Related Factor 2(Nrf2)antioxidant pathway.2018.103:637-644.".Method particularly includes: it uses
Trizol kit (Thermo Fisher Scientific company) extracts total serum IgE.Use one-step method
PrimeScriptTMRNA is synthesized cDNA by RT-PCR Kit (Japanese Takara company).It is template (1 μ L), SYBR by the cDNA
Green Master Mix (Japanese Takara company), specific primer and distilled water are added in PCR amplification system, total volume
50μL。
The specific primer:
Forward primer: 5 '-TCTGAGTCACCTGGACAACC-3 ' (SEQ No.3)；And:
Reverse primer: 5 '-ATCTCAGTGGTATTTGTCAGC-3 ' (SEQ No.4).
PCR program be 95 DEG C 5 minutes, 95 DEG C of 30s, 60 DEG C of 30s, 72 DEG C of 30s, totally 35 circulation.Ago-Gel
Electrophoretic analysis PCR product.In order to confirm ECHS1 gene c.414+5G > A mutation, carried out using standard reagent and condition
Sanger sequencing.
Shearing site analysis:
VariantValidator software (Freeman PJ, Hart RK, Gretton LJ, Brookes AJ,
Dalgleish R.VariantValidator:accurate validation,mapping,and formatting of
sequence variation descriptions.Hum Mutat.2018；39:61-68.) it is used to determine chr10:
G.13345596_13345635 it lacks.
Human Splice Finder software (Desmet FO, Hamroun D, Lalande M, Collod-Beroud
G,Claustres M,Beroud C.Human Splicing Finder:an online bioinformatics tool to
predict splicing signals.Nucleic Acids Res.2009；37:e67), Splice Port software (Dogan
RI,Getoor L,Wilbur WJ,Mount SM.SplicePort—an interactive splice-site
analysis tool.Nucleic Acids Res.2007；35:W285-W291.) and Fruit Fly Splice
Predictor software (The Berkeley Drosophila Genome Project (BDGP) is a consortium of
the Drosophila Genome Center funded by the National Human Genome Research
Institute and the National Institute of General Medical Sciences through its
support of work in the Susan Celniker,J.Ben Brown,Erwin Frise and Gary Karpen
Laboratories.) it is used to the influence of predicted gene shearing site.
Infant gene is analyzed using Sanger sequence, one is found in its ECHS1 [MIM#602292] gene intron 3
The rare mutation of non-coding c.414+5G > A (chr10:g.13345596_13345635 lack GTCAAGAAGCCAGTCATCGC
TGCTGTCAATGGCTATGCC；NM_004092.4).Other regions of genome are without exception.To the genomic DNA from parent
Analysis shows, which is homozygote, and his parent is the heterozygote of the variant.
We utilize Human Splice Finder software, Splice Port software and Fruit Fly Splice
Predictor software carries out bioinformatic analysis to c.414+5G > A mutation first, to predict the function of the mutation.As a result it shows
Show that the mutation likely results in the change of introne splice site, and then the missing of 39bp sequence occurs.Splice Site
Score calculator is used to assess the intensity of splice site, and wild type and the montage of c.414+5G > A saltant type are strong
Degree is respectively 14.2 and 7.9 (http://rulai.cshl.edu/new_alt_exon_db2/html/score.html).(figure
3)
In order to prove that 39bp missing is as caused by detecting c.414+5G > A mutation, we are analyzed using minigene
(minigene analysis) observes base by the PCR amplification and Polyacrylamide Gel Electrophoresis of plasmid specific primer
Because of montage product.As the result is shown: c.414+5G > A mutation has a significant impact (Fig. 4) to the splice mode of ECHS1 gene.Meanwhile
Sanger sequencing result also strong proof, from building c.414+5G > A mutant plasmid in the cDNA segment that obtains also have
The missing of 39bp, it is consistent with the testing result in infant patients cDNA.
C.414+5g > a protein function analysis of variance
Influence of the caused 39bp missing to protein function is mutated by c.414+5G > A in order to detect, we carry out first
Silicon analysis.The result shows that 39bp missing may result in the missing of ECHS1 core enzyme 13 amino acid of catalytic domain.With
Afterwards, three-dimensional structure prediction is carried out to ECHS1 using PyMol software.As a result as Fig. 5 shows that 13 amino acid deletions are located at albumen
Matter active site.C.414+5G > A mutation will lead to the highly conserved sequential amino acid deletion in enzyme activity area, in this way may be very big
Possibility influence ECHS1 protein activity.
The test of ECHS1 gene function:
Based on above-mentioned computer forecast, continue to observe ECHS1 c.414+5G > A is mutated definite influence on its enzyme activity, I
Detect whether infant 2- enoyl-CoA hydratase activity changes.
Primary myoblasts are established from subjects bones' Muscle biospy, are adding 20% (V/V) fetal calf serum (FBS, Thermo
Fisher Scientifific provide) DMEM/F-12 culture medium (Thermo Fisher Scientifific offers) in train
It supports.Condition of culture is 37 DEG C, 5%CO2.
2- enoyl-CoA hydratase, that is, ECHS1 enzyme activity, by spectrophotometry to fibroblast lysate and not
The variation of saturation crotonyl-CoA substrate reactions enough time (15 minutes) absorption photometric afterwards measures.Specific method is shown in Fong
JC,Schulz H.Purification and properties of pig heart crotonase and the
presence of short chain and long chain enoyl coenzyme A hydratases in pig and
guinea pig tissues.J Biol Chem 1977；252:542–Luis PB,Ruiter JP,Ofman R,et
al.Valproic acid utilizes the isoleucine breakdown pathway for its complete
beta-oxidation.Biochem Pharmacol 2011；82:1740–1746.
The result shows that: compared with wild type control, the activity of 2- enoyl-CoA hydratase is significant in infant fibroblast
It reduces, as shown in Figure 6.
As it will be easily appreciated by one skilled in the art that the foregoing is merely illustrative of the preferred embodiments of the present invention, not to
The limitation present invention, any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should all include
Within protection scope of the present invention.
Claims (10)
1. a kind of mitochondrial encephalomyopathy diagnostic kit, which is characterized in that including ECHS1 gene order detection reagent.
2. mitochondrial encephalomyopathy diagnostic kit as described in claim 1, which is characterized in that including ECHS1 gene intron 3
Gene order detection reagent.
3. mitochondrial encephalomyopathy diagnostic kit as claimed in claim 2, which is characterized in that including ECHS1 gene intron 3
Rare mutation c.414+5G > detection reagent of A.
4. mitochondrial encephalomyopathy diagnostic kit as described in claim 1, which is characterized in that the ECHS1 gene order inspection
Test agent includes ECHS1 gene amplification reagent.
5. mitochondrial encephalomyopathy diagnostic kit as claimed in claim 4, which is characterized in that the ECHS1 gene magnification examination
Agent includes:
Forward primer: 5 '-AGCTACACCTGGAGCCACTG-3 '；And:
Reverse primer: 5 '-CCATGGACACGAATCACAAG-3 '.
6. mitochondrial encephalomyopathy diagnostic kit as described in claim 1, which is characterized in that examined including ECHS1 gene expression
Test agent.
7. mitochondrial encephalomyopathy diagnostic kit as claimed in claim 6, which is characterized in that the ECHS1 gene expression inspection
Test agent includes ECHS1 gene RT-PCR primer:
Forward primer: 5 '-TCTGAGTCACCTGGACAACC-3 '；And:
Reverse primer: 5 '-ATCTCAGTGGTATTTGTCAGC-3 '.
8. a kind of application of ECHS1 gene order detection reagent, which is characterized in that it is applied to mitochondrial encephalomyopathy diagnostic reagent
Preparation.
9. application as claimed in claim 8, which is characterized in that it is applied to the abnormal caused mitochondria brain of ECHS1 gene intron 3
The preparation of myopathy diagnostic reagent.
10. application as claimed in claim 9, which is characterized in that c.414+ it is applied to the rare mutation of ECHS1 gene non-coding region
The preparation of mitochondrial encephalomyopathy diagnostic reagent caused by 5G > A.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100113286A1 (en) * | 2007-05-05 | 2010-05-06 | Gilles Andre Lajoie | Methods for detection of preeclampsia |
| CA2881321A1 (en) * | 2012-09-05 | 2014-03-13 | A1M Pharma Ab | Alpha-1-microglobulin for use in the treatment of mitochondria-related diseases |
| CN104789572A (en) * | 2015-05-06 | 2015-07-22 | 南京医科大学 | GPRASP2 mutant type gene as well as identification method and detection kit thereof |
| CN205528786U (en) * | 2016-03-10 | 2016-08-31 | 上海睿玻生物科技有限公司 | Mitochondria brain myopathy, lactic acidosis and palsy appearance disease gene sequencing kit |
| CN107447041A (en) * | 2017-09-28 | 2017-12-08 | 郑州大学第附属医院 | Glioblastoma united diagnostic reagent box and its application method based on CCL8, CCL26 and CXCL6 gene |
| WO2019142845A1 (en) * | 2018-01-17 | 2019-07-25 | 株式会社カネカ | Transformed microorganism for producing pha copolymer comprising 3hh monomer unit at high composition rate and method for producing pha using same |
-
2019
- 2019-09-23 CN CN201910897061.5A patent/CN110452980B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100113286A1 (en) * | 2007-05-05 | 2010-05-06 | Gilles Andre Lajoie | Methods for detection of preeclampsia |
| CA2881321A1 (en) * | 2012-09-05 | 2014-03-13 | A1M Pharma Ab | Alpha-1-microglobulin for use in the treatment of mitochondria-related diseases |
| CN104661673A (en) * | 2012-09-05 | 2015-05-27 | A1M制药公司 | Alpha-1-microglobulin for use in the treatment of mitochondria-related diseases |
| CN104789572A (en) * | 2015-05-06 | 2015-07-22 | 南京医科大学 | GPRASP2 mutant type gene as well as identification method and detection kit thereof |
| CN205528786U (en) * | 2016-03-10 | 2016-08-31 | 上海睿玻生物科技有限公司 | Mitochondria brain myopathy, lactic acidosis and palsy appearance disease gene sequencing kit |
| CN107447041A (en) * | 2017-09-28 | 2017-12-08 | 郑州大学第附属医院 | Glioblastoma united diagnostic reagent box and its application method based on CCL8, CCL26 and CXCL6 gene |
| WO2019142845A1 (en) * | 2018-01-17 | 2019-07-25 | 株式会社カネカ | Transformed microorganism for producing pha copolymer comprising 3hh monomer unit at high composition rate and method for producing pha using same |
Non-Patent Citations (11)
| Title |
|---|
| L. SHI等: "Identification of genetic associations of ECHS1 gene with milk fatty acid traits in dairy cattle", 《ANIMAL GENETICS》 * |
| L. SHI等: "Identification of genetic associations of ECHS1 gene with milk fatty acid traits in dairy cattle", 《ANIMAL GENETICS》, 15 January 2019 (2019-01-15), pages 1 - 9 * |
| TOBIAS B. HAACK等: "Deficiency of ECHS1 causes mitochondrial encephalopathy with cardiac involvement", 《ANNALS OF CLINICAL AND TRANSLATIONAL NEUROLOGY》 * |
| TOBIAS B. HAACK等: "Deficiency of ECHS1 causes mitochondrial encephalopathy with cardiac involvement", 《ANNALS OF CLINICAL AND TRANSLATIONAL NEUROLOGY》, vol. 2, no. 5, 31 December 2015 (2015-12-31), pages 492 * |
| WU ET AL.: "Two novel ECHS1 variants, affecting splicing and reducing enzyme activity, is associated with mitochondrial encephalopathy in infant: a case report", 《BMC NEUROLOGY》 * |
| WU ET AL.: "Two novel ECHS1 variants, affecting splicing and reducing enzyme activity, is associated with mitochondrial encephalopathy in infant: a case report", 《BMC NEUROLOGY》, vol. 20, no. 165, 30 April 2020 (2020-04-30), pages 1 - 8 * |
| 宋艳梅: "线粒体脑肌病的基因突变研究", 中国优秀硕士学位论文全文数据库 医药卫生科技辑, pages 070 - 186 * |
| 张峻滔等: "线粒体脑肌病的治疗进展", 《广东医学》 * |
| 张峻滔等: "线粒体脑肌病的治疗进展", 《广东医学》, no. 24, 25 December 2015 (2015-12-25), pages 147 - 149 * |
| 时雯雯等: "线粒体脑病分子遗传学基础研究若干新进展", 《生命科学》 * |
| 时雯雯等: "线粒体脑病分子遗传学基础研究若干新进展", 《生命科学》, no. 08, 31 December 2017 (2017-12-31), pages 30 - 37 * |
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