CN1563421A - Polymorphism detection chip for gene of enzyme relevant to ethanol metabolism, preparation method of usage - Google Patents
Polymorphism detection chip for gene of enzyme relevant to ethanol metabolism, preparation method of usage Download PDFInfo
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- CN1563421A CN1563421A CN 200410017946 CN200410017946A CN1563421A CN 1563421 A CN1563421 A CN 1563421A CN 200410017946 CN200410017946 CN 200410017946 CN 200410017946 A CN200410017946 A CN 200410017946A CN 1563421 A CN1563421 A CN 1563421A
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Abstract
This invention discloses as alcohol metabolic zyme correlated gene pleiomorphy test chip and its preparation method and usage, designing the following probes on vectors: ADH3: 5'-AAG GTA AAA CAT TTG TTAT5'-AAG GTA AAA AAT TTG TTAT 5'-AAG GTA AAA AAT TTG TTAT5'-AAG CAT AAA GAT TTG TTATA DH2 (allelic gene 1 and allelic gene 2): 5'-AAT CTG TCG CAC AGA TG 5'-AAG GTA AAA CAT TTG TTAT5'-AAT CTG TCC CAC AGA TG5'-AAT CTG TCT CAC AGA TGA DH2(allelic gene 1 and allelic gene 3): 5'-GCA GTA TCT GTA CCG TC CCG TC5'-GCA GTA TCG CTA CCG TC ALDH2:5'-CAT ACA CTG AAG TGA AA5'-CAT ACA CTA AAG TGA AA5'-CAT ACA CTT AAG TGA AA 5'-CAT ACA CTC AAG TGAAA. The advantage is that it realizes quick, simple and high flux test to alcohol metabolic zyme correlated gene ADH2, ADH3 and ALDH2 pleiomorphy.
Description
The present invention relates to a kind of alcohol metabolism relative enzyme gene polymorphic test chips and its preparation method and application for technical field.
Background technique
Alcohol abuse and alcohol dependence have become the more worth people of the public health problem got worse in the world today, especially adolescents alcohol and pay attention to.The alcoholic liver disease past is more rare in China, therefore it is also few for studying it, but the disease incidence of alcoholic liver disease obviously increases in recent years, it has also become is only second to the second largest hepatopathy of virus hepatitis.Alcohol has an apparent toxic effect to liver, and 80% or more has a degree of fatty liver in heavy drinking essence person, and 10%-35% can develop into alcoholic hepatitis, and 10%-20% would develop into cirrhosis.
Metabolic conversion is acetaldehyde and acetic acid to alcohol (ethyl alcohol) in vivo, and predominantly alcohol dehydrogenase (alcoholdehydrogenase, ADH) and acetaldehyde dehydrogenase (aldehyde dehydrogenase, ALDH) acts on.The isodynamic enzyme ADH1 (α) that ADH has 3 kinds of different genes to encode, ADH2 (β), ADH3 (γ), there are certain polymorphisms by ADH2 and ADH3 in base, there are three allele sites by ADH2, ADH2*1, ADH2*2 and ADH2*3, it is separately encoded β 1, 3 three subunits of β 2 and β, the polymorphism of ADH2 allele is the single base replacement due to 47 (G ψ A) and 369 (C ψ T) codons:, the isoenzymes of β 1 has low Km (0.049mmol/L) and low maximal clearance (Vmax) (0.23U/min) to alcohol, the isoenzymes of β 2. compares with low Km (0.94mmol/L) maximal clearance (8.6U/min), isoenzymes β 3 With to the high Km of alcohol (36mmol/L) and high Vmax (7.9U/min).The activity of ADH3*1 expression product is 2 times of ADH3*2.The allele that ALDH2 gene encodes active and inactive subunit is named as ALDH2*1 and ALDH2*2 respectively, the difference of these isoenzymes is that the base of 487 codons exchanges (G ψ A), generates the exchange of lysine (inactive subunit) and glutamic acid (active subunit).The polymorphism of Alcohol-metabolizing enzymes related gene plays an important role in alcoholic liver disease and other metabolism and the morbidity of drug-induced liver disease.
Doing genetic analysis for SNPs is to refer in particular to a large amount of existing single nucleotide variations in detection human genome, this is related [1] with disease susceptibility and poisonous substance neurological susceptibility.SNPs is not only the label of human race and individual difference, is also the label for determining different crowd race and individual difference, can become and find disease related gene, carry out the basis of medical diagnosis on disease, prevention and drug screening.Conventional method for SNPs detection has allele specific oligonucleotide (ASO) hybridization, restriction enzyme enzyme process (RFLP), site-specific primer PCR (PCR-SSP) and direct Sequencing etc..The development of biochip technology provides possibility [2] for the detection method of new SNPs.Oligonucleotide microarray is a kind of technique of gene detection that newly-developed gets up, a large amount of oligonucleotide probe can be regularly arranged on a slide by it, these probes can be combined with the complementary series of the extension increasing sequence of the sample DNA of signal display mass signatures, by being detected to semiochemicals, software processing analysis is carried out to results of hybridization, to obtain the intensity and its distribution pattern figure of hybridization signal.
Bibliography:
[1] Khner M K, Beerli P, Yamato J, et al.Usefulness of single nucleatide polymorphismdata for estimating population parameters.Genetics, 2000,156:439-447
[2] Hirschlhorn J N, Sklar P, Lindblad-Toh K, et al.SBE-TAGS:an array-based methodfor efficient single-nucleotide polymorphism genotyping.Proc Natl Acad SciUSA, 2000,97:12164-12169
[3] Picoult-Newberg L, Idker TE, Pohl M G, et al.Mining SNPs from ESTdatabases.Genome Res, 1999,9:167-174.
[4] Yu Chaohui Chen Lihua, the preparation and application China digestion magazine 2003,23 (11) of the alcohol metabolizing enzyme oligonucleotide genetic chip such as Chen Weixing；695-6
Summary of the invention
The object of the present invention is to provide a kind of alcohol metabolism relative enzyme gene polymorphic test chips and its preparation method and application.
Alcohol metabolism relative enzyme gene polymorphic test chip: following probe is equipped on carrier:
ADH3:5 '-AAG GTA AAA CAT TTG TTAT
5’-AAG GTA AAA TAT TTG TTAT
5’-AAG GTA AAA AAT TTG TTAT
5’-AAG GTA AAA GAT TTG TTAT
ADH2 (allele 1 and allele 2): 5 '-AAT CTG TCG CAC AGA TG
5’-AAG GTA AAA CAT TTG TTAT
5’-AAT CTG TCC CAC AGA TG
5’-AAT CTG TCT CAC AGA TG
ADH2 (allele 1 and allele 3): 5 '-GCA GTA TCT GTA CCG TC
5’-GCA GTA TCC GTA CCG TC
5’-GCA GTA TCA GTA CCG TC
5’-GCA GTA TCG GTA CCG TC
ALDH2:5 '-CAT ACA CTG AAG TGA AA
5’-CAT ACA CTA AAG TGA AA
5’-CAT ACA CTT AAG TGA AA
5’-CAT ACA CTC AAG TGA AA
Alcohol metabolism relative enzyme gene polymorphic test chip preparation method step are as follows:
1) processing of chip carrier: wave carrier piece chromic acid lotion soaked overnight, washing, in 24~26% ammonium hydroxide overnight, washing, sheet glass immerses in 95% ethanol solution of aminopropyl trimethoxysilane, adjusts PH to 4~4.5, the cleaning of 95% EtOH Sonicate with glacial acetic acid, 150~160 DEG C dry 3~5 hours, and 9~11% glutaraldehydes are processed into aldehyde radical；
2) synthesis of primer and probe: a set of probe (such as table 1) is devised for Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; 4 pairs of primers (such as table 2) have been synthesized for the design of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; respectively there is a Cyp-3 signaling molecule label in each pair of primer; with 75~85 DEG C of concentrated ammonia liquor deprotections after synthesis; cutting 14~17 hours; OPC column purification; it is ultraviolet it is quantitative after be dried in vacuo concentration, -20~-80 DEG C save backup；
3) genetic chip preparation and post-processing: oligonucleotide probe is dissolved in 3 × SSC solution with 15 μM of concentration, and oligonucleotides is sprayed on the slide of aldehyde radical with PixySys5500 chip preparing instrument, and point spacing is 0.5mm.After point sample, left at room temperature over night.
Alcohol metabolism relative enzyme gene polymorphic test chip is used for the polymorphic detection and parting of patients with alcoholic liver disease Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2.
The present invention is by the probe spotting Jing Guo selection on slide, hybridized respectively with the DNA fragmentation of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 through PCR amplification, disposably obtain the polymorphic situation in each site, quick, simple and direct, high-throughput detection and parting are carried out to Alcohol-metabolizing enzymes related gene polymorphism to realize, there is important social and economic benefit.
Specific embodiment
Immobilised probe is oligonucleotide probe on slide of the present invention, its sequence is designed according to the gene expression characteristics of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2, plays length range in 15-20 base, generally 18 bases, its Tm value is almost the same, at 40~43 DEG C.
Described primer is designed according to the gene expression characteristics of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2, and primer sequence length is respectively used to the amplification of said gene between 20-24 base.
Signaling molecule label is the primer expanded for the gene order of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2.Signaling molecule label is fluorescent molecule.
Table 1: the probe of alcohol metabolism relative enzyme gene polymorphic test chip
| gene | probe |
| ADH3 | 5’-AAG GTA AAA CAT TTG TTAT |
| 5’-AAG GTA AAA TAT TTG TTAT | |
| 5’-AAG GTA AAA AAT TTG TTAT | |
| 5’-AAG GTA AAA GAT TTG TTAT | |
| ADH2 (allele 1 and allele 2) | 5’-AAT CTG TCG CAC AGA TG |
| 5’-AAT CTG TCA CAC AGA TG | |
| 5’-AAT CTG TCC CAC AGA TG | |
| 5’-AAT CTG TCT CAC AGA TG | |
| ADH2 (allele 1 and allele 3) | 5’-GCA GTA TCT GTA CCG TC |
| 5’-GCA GTA TCC GTA CCG TC | |
| 5’-GCA GTA TCA GTA CCG TC | |
| 5’-GCA GTA TCG GTA CCG TC | |
| ALDH2 | 5’-CAT ACA CTG AAG TGA AA |
| 5’-CAT ACA CTA AAG TGA AA | |
| 5’-CAT ACA CTT AAG TGA AA | |
| 5’-CAT ACA CTC AAG TGA AA |
Table 2: the primer of alcohol metabolism relative enzyme gene polymorphic test chip
| Gene | Cyp-3 fluorescent dye primer | Primer |
| ADH3 | 5’-CTT TAA GAG TAA AGA ATC TGT CC-3’ | 5’-ACC TCT TTC CAG AGC GAAGCAG-3’ |
| ADH2 (allele 1 and allele 2) | 5’-GAA GGG GGG TCA CCA GGT TGC-3’ | 5’-AAT CTT TTC TGA ATCTGA ACAG-3’ |
| ADH2 (allele 1 and allele 3) | 5’-TTG ATA ACA TCT CTG AAG AGC TGA-3’ | 5’-TGG ACT CTC ACA ACA AGCATGT-3’ |
| ALDH2 | 5’-CAG GTC CCA CAC TCA CAG TTT-3’ | 5’-CAC CCT TTG GTG GCT ACAAG-3’ |
Embodiment 1: the preparation of nucleotide gene chip:
1) processing of chip carrier: wave carrier piece chromic acid lotion soaked overnight is washed, in 25% ammonium hydroxide overnight, washing.Sheet glass immerses in 95% ethanol solution of aminopropyl trimethoxysilane, adjusts PH to 4.5 with glacial acetic acid, the cleaning of 95% EtOH Sonicate, 160 DEG C dry 4 hours, and 5% glutaraldehyde is processed into aldehyde radical.
2) synthesis of primer and probe: the present invention devises a set of probe (being shown in Table 1) for Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene.4 pairs of primers (being shown in Table 2) have been synthesized for the design of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene.Respectively there is a Cyp-3 fluorescent marker in each pair of primer.With 80 DEG C of concentrated ammonia liquor deprotections after synthesis, cut 15 hours, OPC column purification.It is ultraviolet it is quantitative after be dried in vacuo concentration, -20 DEG C save backup.
3) genetic chip preparation and post-processing: oligonucleotide probe is dissolved in 3 × SSC solution with 15 μM of concentration, and oligonucleotides is sprayed on the slide of aldehyde radical with PixySys5500 chip preparing instrument, and point spacing is 0.5mm.After point sample, left at room temperature over night.
Embodiment 2: the preparation of nucleotide gene chip:
1) processing of chip carrier: wave carrier piece chromic acid lotion soaked overnight, washing, in 24% ammonium hydroxide overnight, washing, sheet glass immerses in 95% ethanol solution of aminopropyl trimethoxysilane, adjusts PH to 4, the cleaning of 95% EtOH Sonicate with glacial acetic acid, 150 DEG C dry 3 hours, and 9% glutaraldehyde is processed into aldehyde radical；
2) synthesis of primer and probe: a set of probe is devised for Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; 4 pairs of primers have been synthesized for the design of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; respectively there is a Cyp-3 signaling molecule label in each pair of primer; with 75 DEG C of concentrated ammonia liquor deprotections after synthesis; cutting 13 hours; OPC column purification, it is ultraviolet it is quantitative after be dried in vacuo concentration, -20 DEG C save backup；
3) genetic chip preparation and post-processing: oligonucleotide probe is dissolved in 3 × SSC solution with 15 μM of concentration, and oligonucleotides is sprayed on the slide of aldehyde radical with PixySys5500 chip preparing instrument, and point spacing is 0.5mm.After point sample, left at room temperature over night.
The preparation of 3 nucleotide gene chip of embodiment:
1) processing of chip carrier: wave carrier piece chromic acid lotion soaked overnight, washing, in 26% ammonium hydroxide overnight, washing, sheet glass immerses in 95% ethanol solution of aminopropyl trimethoxysilane, adjusts PH to 4.5, the cleaning of 95% EtOH Sonicate with glacial acetic acid, 160 DEG C dry 5 hours, and 11% glutaraldehyde is processed into aldehyde radical；
2) synthesis of primer and probe: a set of probe is devised for Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; 4 pairs of primers have been synthesized for the design of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; respectively there is a Cyp-3 signaling molecule label in each pair of primer; with 85 DEG C of concentrated ammonia liquor deprotections after synthesis; cutting 17 hours; OPC column purification, it is ultraviolet it is quantitative after be dried in vacuo concentration, one 80 DEG C save backup；
3) genetic chip preparation and post-processing: oligonucleotide probe is dissolved in 3 × SSC solution with 15 μM of concentration, and oligonucleotides is sprayed on the slide of aldehyde radical with PixySys5500 chip preparing instrument, and point spacing is 0.5mm.After point sample, left at room temperature over night.
The application method of alcohol metabolism relative enzyme gene polymorphic test chip are as follows:
1) slide for being loaded with oligonucleotide probe is respectively washed twice using preceding 0.2%SDS and clear water, it is air-dried rear 1%NaBH4 solution reduction 10min, 0.2%SDS is washed once, and washing is primary, with for use after being air-dried, complete the covalent immobilization of probe in surface of glass slide.
2) contain 50ng DNA, 4 pairs of primers (table 2), 10mM Tris-HClpH8.3,50mM KCl, 1.5mM MgCl in 20ul PCR reaction system2, 200uM dNTP and 1.5 unit Taq enzymes；Wherein primer content is 20ng Cyp-3 fluorescent dye primer, 2ng general primer.PCR cycle condition: 95 DEG C of denaturation in 5 minutes；94 DEG C of denaturation of each 45 seconds of 35 circulations, 57.5 DEG C of annealing, 72 DEG C of extensions；It is finally 72 DEG C of extensions in 5 minutes.
3) after 2ulPCR product is mixed with 10ul hybridization buffer, hybridize 1 hour with the oligonucleotide probe slide prepared at 42 DEG C, then respectively with 1 × SSC, 0.2%SDS, 0.2 × SSC, 0.1 × SSC cleaning, it is finally detected with laser co-focusing Fluorescence Scanner ScanArray 3000, is scanned 3 times under 85% laser intensity.Positive control probe composition is analyzed, judges the differentiation of the gene pleiomorphism of alcohol metabolism relevant enzyme.Alcohol metabolism relative enzyme gene polymorphic test chip uses example: to 165 wine-heads, wherein 43 without liver damage, it is alcoholic liver disease 122, the age 25~70 years old, 44.0 years old average.It is Normal group 65, the age 26~68 years old, 45.9 years old average.All research objects are all from Zhejiang Province Epidemiology of alcoholic liver diseases survey group.Diagnostic criteria excludes virus B hepatitis surface antigen and viral hepatitis type C antibody positive referring to " diagnosis basis of alcoholic liver disease and healing, improvement standard [3] ", and the person of not being true to type does needle biopsy of liver.All objects extract 3ml peripheral blood, routinely extract DNA and are detected with the said chip above method.As a result are as follows: 1) ADH2 gene polynorphisms: healthy control group, wine-head's group, the ADH2*1 gene frequency without liver damage group and alcoholic liver disease group are respectively 37.69%, 55.76%, 46.51%, 59.02%；ADH2*2 gene frequency is respectively 62.31%, 44.24%, 53.49%, 40.98%；ADH2*3 allele is not found.The ADH2*1 frequency of wine-head and patients with alcoholic liver disease is apparently higher than healthy control group, and patients with alcoholic liver disease is higher than the wine-head without liver damage；2) ADH3 gene pleiomorphism: for the gene frequency of the ADH3 gene of wine-head and alcoholic liver disease compared with healthy control group, the ADH3*2 frequency of the wine-head of no liver damage is higher than normal healthy controls.3) ALDH2 gene pleiomorphism: the ALDH2*2 gene frequency of wine-head and patients with alcoholic liver disease is lower than healthy control group, and patients with alcoholic liver disease ALDH2*2 gene frequency is substantially less than the wine-head [4] without liver damage.
Claims (9)
1, a kind of alcohol metabolism relative enzyme gene polymorphic test chip, it is characterised in that: following probe is equipped on carrier:
ADH3:5 '-AAG GTA AAA CAT TTG TTAT
5’-AAG GTA AAA TAT TTG TTAT
5’-AAG GTA AAA AAT TTG TTAT
5’-AAG GTA AAA GAT TTG TTAT
ADH2 (allele 1 and allele 2): 5 '-AAT CTG TCG CAC AGA TG
` 5’-AAG GTA AAA CAT TTG TTAT
5’-AAT CTG TCC CAC AGA TG
5’-AAT CTG TCT CAC AGA TG
ADH2 (allele 1 and allele 3): 5 '-GCA GTA TCT GTA CCG TC
5’-GCA GTA TCC GTA CCG TC
5’-GCA GTA TCA GTA CCG TC
5’-GCA GTA TCG GTA CCG TC
ALDH2:5 '-CAT ACA CTG AAG TGA AA
5’-CAT ACA CTA AAG TGA AA
5’-CAT ACA CTT AAG TGA AA
5’-CAT ACA CTC AAG TGA AA
2, a kind of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 1, it is characterised in that: described carrier is slide.
3, a kind of preparation method of alcohol metabolism relative enzyme gene polymorphic test chip, it is characterised in that: method and step are as follows:
1) processing of chip carrier: wave carrier piece chromic acid lotion soaked overnight, washing, in 24~26% ammonium hydroxide overnight, washing, slide immerses in 95% ethanol solution of aminopropyl trimethoxysilane, adjusts PH to 4~4.5, the cleaning of 95% EtOH Sonicate with glacial acetic acid, 150~160 DEG C dry 3~5 hours, and 9~11% glutaraldehydes are processed into aldehyde radical；
2) synthesis of primer and probe: a set of probe is devised for Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; 4 pairs of primers have been synthesized for the design of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2 gene; respectively there is a Cyp-3 signaling molecule label in each pair of primer; with 75~85 DEG C of concentrated ammonia liquor deprotections after synthesis; cutting 13~17 hours; OPC column purification, it is ultraviolet it is quantitative after be dried in vacuo concentration, -20~-80 DEG C save backup；
3) genetic chip preparation and post-processing: oligonucleotide probe is dissolved in 3 × SSC solution with 15 μM of concentration, and oligonucleotides is sprayed on the slide of aldehyde radical with PixySys5500 chip preparing instrument, and point spacing is 0.5mm.After point sample, left at room temperature over night immobilization.
4, the preparation method of a kind of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 3, it is characterized by: immobilised probe is oligonucleotide probe on described slide, its sequence is designed according to the gene expression characteristics of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2, for its length range in 15-20 base, Tm value is 40~43 DEG C.
5, the preparation method of a kind of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 3, it is characterised in that: described probe is:
ADH3:5 '-AAG GTA AAA CAT TTG TTAT
5’-AAG GTA AAA TAT TTG TTAT
5’-AAG GTA AAA AAT TTG TTAT
5’-AAG GTA AAA GAT TTG TTAT
ADH2 (allele 1 and allele 2): 5 '-AAT CTG TCG CAC AGA TG
5’-AAG GTA AAA CAT TTG TTAT
5’-AAT CTG TCC CAC AGA TG
5’-AAT CTG TCT CAC AGA TG
ADH2 (allele 1 and allele 3): 5 '-GCA GTA TCT GTA CCG TC
5’-GCA GTA TCC GTA CCG TC
5’-GCA GTA TCA GTA CCG TC
5’-GCA GTA TCG GTA CCG TC
ALDH2:5 '-CAT ACA CTG AAG TGA AA
5’-CAT ACA CTA AAG TGA AA
5’-CAT ACA CTT AAG TGA AA
5’-CAT ACA CTC AAG TGA AA
6, the preparation method of a kind of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 3, it is characterized by: described primer is that root one is designed according to the gene expression characteristics of Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2, primer sequence length is respectively used to the amplification of said gene between 20-24 base.
7, a kind of preparation method of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 3, it is characterised in that: described primer are as follows:
Gene C yp-3 fluorescent dye primer primer
ADH3 5’-CTT TAA GAG TAA AGA ATC TGT CC-3’ 5’-ACC TCT TTC CAG
AGC GAA GCAG-3’
ADH2 5’-GAA GGG GGG TCA CCA GGT TGC-3’ 5’-AAT CTT TTC TGA
ATC TGA ACAG-3’
(allele 1 and allele 2)
ADH2 5’-TTG ATA ACA TCT CTG AAG AGC TGA-3’ 5’-TGG ACT CTC ACA
ACA AGC ATGT-3’
(allele 1 and allele 3)
ALDH2 5’-CAG GTC CCA CAC TCA CAG TTT-3’ 5’-CAC CCT TTG GTG
GCT ACA AG-3’
8, the preparation method of a kind of alcohol metabolism relative enzyme gene polymorphic test chip according to claim 3, it is characterised in that: described signaling molecule label is fluorescent molecule.
9, a kind of purposes of alcohol metabolism relative enzyme gene polymorphic test chip, it is characterised in that polymorphic detection and parting for patients with alcoholic liver disease Alcohol-metabolizing enzymes related gene ADH2, ADH3 and ALDH2.
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| CN1268769C CN1268769C (en) | 2006-08-09 |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007039550A3 (en) * | 2005-09-29 | 2007-08-09 | Proyecto Biomedicina Cima Sl | Molecular markers of hepatocellular carcinoma and their applications |
| CN102146438A (en) * | 2010-12-22 | 2011-08-10 | 协和干细胞基因工程有限公司 | Kit for detecting alcoholic liver disease susceptibility |
| CN102549171A (en) * | 2009-10-07 | 2012-07-04 | 学校法人武库川学院 | Genotype determination method |
| CN103361432A (en) * | 2013-07-26 | 2013-10-23 | 天津市秀鹏生物技术开发有限公司 | Primer group and kit for detecting genetic typing of human aldehyde dehydrogenase 2 (ALDH2) |
| CN103540675A (en) * | 2013-10-31 | 2014-01-29 | 上海百傲科技股份有限公司 | Specific primer pair and probe for detecting ALDH2 (alcohol dehydrogenase 2) gene chip |
| CN103834733A (en) * | 2014-02-27 | 2014-06-04 | 厦门大学附属中山医院 | Kit and method for determining mutation sites of genes of acetaldehyde dehydrogenase 2 and methylene tetrahydrofolic acid reductase by virtue of single tube at the same time |
| CN104156632A (en) * | 2014-07-21 | 2014-11-19 | 金华市中心医院 | Method for intelligently designing drug metabolic enzyme detecting gene chip probes |
| CN113774115A (en) * | 2021-08-04 | 2021-12-10 | 上海百傲科技股份有限公司 | Method, kit, primer pair, probe, gene chip and application for detecting ALDH2 gene |
-
2004
- 2004-04-21 CN CN 200410017946 patent/CN1268769C/en not_active Expired - Lifetime
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007039550A3 (en) * | 2005-09-29 | 2007-08-09 | Proyecto Biomedicina Cima Sl | Molecular markers of hepatocellular carcinoma and their applications |
| CN102549171A (en) * | 2009-10-07 | 2012-07-04 | 学校法人武库川学院 | Genotype determination method |
| CN102146438A (en) * | 2010-12-22 | 2011-08-10 | 协和干细胞基因工程有限公司 | Kit for detecting alcoholic liver disease susceptibility |
| CN103361432A (en) * | 2013-07-26 | 2013-10-23 | 天津市秀鹏生物技术开发有限公司 | Primer group and kit for detecting genetic typing of human aldehyde dehydrogenase 2 (ALDH2) |
| CN103540675A (en) * | 2013-10-31 | 2014-01-29 | 上海百傲科技股份有限公司 | Specific primer pair and probe for detecting ALDH2 (alcohol dehydrogenase 2) gene chip |
| CN103834733A (en) * | 2014-02-27 | 2014-06-04 | 厦门大学附属中山医院 | Kit and method for determining mutation sites of genes of acetaldehyde dehydrogenase 2 and methylene tetrahydrofolic acid reductase by virtue of single tube at the same time |
| CN104156632A (en) * | 2014-07-21 | 2014-11-19 | 金华市中心医院 | Method for intelligently designing drug metabolic enzyme detecting gene chip probes |
| CN113774115A (en) * | 2021-08-04 | 2021-12-10 | 上海百傲科技股份有限公司 | Method, kit, primer pair, probe, gene chip and application for detecting ALDH2 gene |
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| Publication number | Publication date |
|---|---|
| CN1268769C (en) | 2006-08-09 |
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