CN106834458A - The method of the homogeneous exons mutation index of detection EGFR allele 19 - Google Patents
The method of the homogeneous exons mutation index of detection EGFR allele 19 Download PDFInfo
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Abstract
The present invention relates to the method for the homogeneous exons mutation index of detection EGFR allele 19, particular agent is expanded, then amplified production row FAM and R110 fluorescence polarization value are detected, according to the height change of FAM and R110 fluorescence polarization values and the ratio interpretation exons mutation index of sample EGFR allele 19 to be measured.The present invention creatively proposes the exons mutation index of EGFR allele 19 and the method detected to the exons mutation index of EGFR allele 19, the exons mutation index of EGFR allele 19 is, more sensitive special molecular marker related to drug responsiveness higher degree compared with the Varia nt in exon type absolute quantity of EGFR allele 19, and the exons mutation index of detection EGFR allele 19 is compared with only detecting variation type and the more scientific accurate, criterion and quantity of absolute quantity, the detection for more having predictive value.
Description
Technical field
The present invention relates to the exons mutation index detection technique field of EGFR allele 19, and in particular to a kind of homogeneous inspection
The method for surveying the exons mutation index of EGFR allele 19.
Background technology
Allelic variation is the change for controlling certain character gene in cell in pair of homologous chromosome double-stranded DNA same position
It is different.In genes of individuals group, allelic variation and wild type exist simultaneously, mutually restriction balance, joint effect gene expression and
Function, influences biological respinse, determines that cell lapses to property isophenous；There is heterogeneous and dosage effect in individual internal allelic variation
Should, Different Individual is different from the relative populations level of wild type in different phase allelic variation, and make a variation relative populations level
Change directly affects gene function and expression.The Varia nt in exon of EGFR allele 19 exists simultaneously with wild type, interacts,
Together decide on cell EGFR targeted drugs reactivity phenotype.The extron E746-A750's of Different Individual EGFR allele 19 lacks
Mistake mutation content difference is larger, and the ratio of the Varia nt in exon type of EGFR allele in genome 19 and wild type is phase by we
Quantity levels are named as with the exons mutation index of EGFR allele 19, it is individual compared with the change of the extron of EGFR allele 19
Special-shaped other and absolute quantity is related to drug responsiveness higher degree, more sensitive special, the more scientific accurate, molecule of criterion and quantity
Mark.Therefore the extron wild type of EGFR allele 19 and saltant type relative amount in individuality：The extron of EGFR allele 19
Mutation index is more fully EGFR targeting medicine drug efficacy prediction indexs.But currently available technology or only detection EGFR allele 19
Exons mutation only detects its absolute magnitude, and does not detect the extron wild type of EGFR allele 19 and contain with the relative of saltant type
Amount ratio, because detection object is unilateral, testing result is by factor shadows such as materials genetic mutation heterogeneity, method of drawing material, quantity positions
Ring, application value is low；Or detection method is cumbersome poorly efficient, not standardized.
The content of the invention
The exons mutation index of EGFR allele 19 is homogeneously detected it is an object of the invention to provide a kind of fluorescence polarization
Method, is the extron wild type of EGFR allele 19 and saltant type content for the exons mutation index of EGFR allele 19
The detection of ratio, overcomes the unilateral of prior art presence, high cost, complex operation step, the problem of easily pollution.
The technical solution adopted in the present invention is：
The method of the homogeneous exons mutation index of detection EGFR allele 19, it is characterised in that：
Agents useful for same includes：
MgCl2, 10 × PCR reaction buffers, the extron DNA primer of dNTPs, EGFR allele 19, EGFR equipotential bases
Because of 19 extron E746-A750, the FAM label probes of the 15bp deletion mutations DNA of 2236 to 2250 bit bases, EGFR equipotential bases
Because of 19 extron E746-A750, the R110 label probes of the 15bp wild types DNA of 2236 to 2250 bit bases, template：It is to be detected
Sample DNA, negative control standard items are outer aobvious pGEM-T-easy empty carriers, positive control standard items i.e. EGFR allele 19
The mixture of sub- wild type reference standards and 19 exons mutation type reference standards, concentration is 1000 copies/ml；
Reagent is expanded respectively, the detection of FAM and R110 fluorescence polarization values is then carried out respectively to amplified production, sun
Property reference standards FAM and R110 fluorescence polarization value is respectively greater than negative control standard items FAM and R110 fluorescence polarization value 30mp
Below it is high with the difference of positive and negative reference standards according to detected sample FAM and R110 fluorescence polarization value then to expand successfully
Low change and the exons mutation index of EGFR allele 19 of ratio interpretation detected sample.
Amplified reaction is carried out in 25uL systems, adds the μ L of 10 × PCR reaction buffers 2.5, and concentration is 2.0mmol/L
DGTP, dCTP, dATP and dTTP, archaeal dna polymerase 1-2 units, concentration be 1.5-3.0mM MgCL2.Concentration is 0.5uM's
The extron sense primer of EGFR gene 19 and 3 ' 2 kind probes of the end with fluorescence labeling, the EGFR equipotentials of concentration 0.1uM-0.05uM
The extron anti-sense primer of gene 19；5 μ L template DNAs；
PCR buffer solutions are 1.0U Taq polymerase.
According to EGFR allele GenBank sequences, design synthesis covers the extron E746-A750's of allele containing EGFR 19
Primer and 19 extron wild types, saltant type probe：
EGFR19 forward primers 5'-gtgcatcgctggtaacatcc-3'；
EGFR19 reverse primers 5'-gaagcaacatctccgaaaagc-3'；
19 extron wild-type probes：5’-gagggaattaagagaagctc–R110 3’；
19 exons mutation type probes：5’-gccaaaacatctccggc-FAM 3’.
Amplification reaction condition is：After 94 DEG C of denaturation 5min, 95 DEG C are incubated 1 second, and 60 DEG C are incubated 1 second, and 72 DEG C are incubated 10 seconds, 45
Individual circulation, last circulation is without 72 DEG C of incubations.
The extron negative control standard items template of amplification EGFR allele 19 is pGEM-T-easy empty carriers, sun respectively
Property reference standards template be the extron wild type control standard items 1000 of EGFR allele 19 copy/ml+19 extrons dash forward
Modification reference standards 1000 copy/ml and DNA sample to be measured, the extron genes of interest of EGFR allele 19 is obtained respectively
Amplified production.
Methods described detects negative control standard items, positive control standard items and testing sample EGFR allele 19 respectively
FAM the and R110 fluorescence polarization values of the final amplified production of extron genes of interest；
It is inclined that positive control standard items R110, FAM fluorescence polarization value is respectively greater than negative control standard items R110, FAM fluorescence
More than the value 30mp that shakes is then to expand successfully；
The exons mutation index of EGFR allele 19 of testing sample={ (the FAM fluorescence polarization values of testing sample-(sun
Property reference standards FAM fluorescence polarization values-negative control standard items FAM fluorescence polarization values))/(FAM is glimmering for positive control standard items
Light polarization value-negative control standard items FAM fluorescence polarization values) } ÷ { (the R110 fluorescence polarization values-(positive controls of testing sample
Standard items R110 fluorescence polarization values-negative control standard items R110 fluorescence polarization values))/(positive control standard items R110 fluorescence is inclined
Shake value-negative control standard items R110 fluorescence polarization values) }.
The present invention has advantages below：
The present invention creatively proposes the concept of the exons mutation index of EGFR allele 19 and to EGFR equipotential bases
Because 19 exons mutation indexes carry out fluorescence polarization homogeneously detection method, that is, to EGFR allele 19 in genome outside
Aobvious subbase is the method that relative populations level is detected because of the ratio of anomaly and wild type.Detection EGFR allele 19
Exons mutation index method is anti-with EGFR targeting medicine medicines compared with the detection of the Varia nt in exon absolute quantity of EGFR allele 19
Answering property higher degree is related, more sensitive special, more scientific accurate, criterion and quantity, the molecular marker detection method that more has predictive value.
Compared with prior art, it is an advantage of the invention that：
1st, the concept of the exons mutation index of EGFR allele 19 is more scientific accurate.
2nd, this method step is simple, simple to operate：The extron wild type of EGFR allele 19 and saltant type phase in sample
A step in content detection stopped pipe is carried out, pollution is not susceptible to, as a result accurately.
3rd, interpretation of result is simple：The exons mutation index of EGFR allele 19 is only needed outside EGFR allele 19 when judging
Show sub- wild type to compare with saltant type fluorescence polarization detection numerical calculation, easily standardization, automation；
4th, low cost：Fluorescence polarization is homogeneously detected.Any special agent and fluorescent quenching or very low power bonding agent are not required to, no
Need real-time detection.
5th, have a wide range of application：Can be used to detect cell, blood or tissue specimen.
Specific embodiment
With reference to specific embodiment, the present invention will be described in detail.
The present invention relates to the method for the homogeneous exons mutation index of detection EGFR allele 19, agents useful for same includes：
MgCl2, 10 × PCR reaction buffers, the extron DNA primer of dNTPs, EGFR allele 19, EGFR equipotential bases
Because of 19 extron E746-A750, the FAM label probes of the 15bp deletion mutations DNA of 2236 to 2250 bit bases, EGFR equipotential bases
Because of 19 extron E746-A750, the R110 label probes of the 15bp wild types DNA of 2236 to 2250 bit bases, template：It is to be detected
Sample DNA, negative control standard items (pGEM-T-easy empty carriers), the positive control standard items (extron of EGFR allele 19
The mixture of wild type control standard items and 19 exons mutation type reference standards, concentration is 1000 copies/ml)；
Reagent is expanded respectively, the detection of FAM and R110 fluorescence polarization values is then carried out respectively to amplified production, sun
Property reference standards FAM and R110 fluorescence polarization value is respectively greater than negative control standard items FAM and R110 fluorescence polarization value 30mp
Below it is high with the difference of positive and negative reference standards according to detected sample FAM and R110 fluorescence polarization value then to expand successfully
Low change and the exons mutation index of EGFR allele 19 of ratio interpretation detected sample.
Amplified reaction is carried out in 25uL systems, adds the μ L of 10 × PCR reaction buffers 2.5, and concentration is 2.0mmol/L
DGTP, dCTP, dATP and dTTP, archaeal dna polymerase 1-2 units, concentration be 1.5-3.0mM MgCL2.Concentration is 0.5uM's
The extron sense primer of EGFR gene 19 and 3 ' 2 kind probes of the end with fluorescence labeling, the EGFR equipotentials of concentration 0.1uM-0.05uM
The extron anti-sense primer of gene 19；5 μ L template DNAs.
According to EGFR allele GenBank sequences (AY588246), the extron of allele containing EGFR 19 is covered in design synthesis
The primer of E746-A750 (2235-2249, GGAATTAAGAGAAGC) and 19 extron wild types, saltant type probe：EGFR19
Forward primer 5'-gtgcatcgctggtaacatcc-3'(No.1), EGFR19 reverse primers 5'-
Gaagcaacatctccgaaaagc-3'(No.2), 19 extron wild-type probe：5’-gagggaattaagagaagctc–
R110 3 ' (No.3), 19 exons mutation type probes：5’-gccaaaacatctccggc-FAM 3’(No.4).
Amplification reaction condition is：After 94 DEG C of denaturation 5min, 95 DEG C are incubated 1-20 seconds, and 55-60 DEG C is incubated 1-20 seconds, and 72 DEG C incubate
Educate 10-40 seconds, 45 circulations, last circulation is without 72 DEG C of incubations.
In method, the extron negative control standard items template of EGFR allele 19 is expanded respectively, and (pGEM-T-easy is unloaded
Body), positive control standard items template (the extron wild type control standard items 1000 of EGFR allele 19 copy/ml+19 outside show
Sub- saltant comparison standard items 1000 copy/ml) and DNA sample to be measured, the extron purpose of EGFR allele 19 is obtained respectively
Gene amplification product.
The method of the homogeneous exons mutation index of detection EGFR allele 19 of the present invention, it is adaptable to fluorescence polarization
Detection technique.Negative control standard items, positive control standard items and the extron of testing sample EGFR allele 19 are detected respectively
FAM the and R110 fluorescence polarization values (mp) of the final amplified production of genes of interest.Positive control standard items R110, FAM fluorescence is inclined
Value of shaking is respectively greater than more than negative control standard items R110, FAM fluorescence polarization value 30mp then to expand successfully.
The exons mutation index of EGFR allele 19 of testing sample={ (the FAM fluorescence polarization values of testing sample-(sun
Property reference standards FAM fluorescence polarization values-negative control standard items FAM fluorescence polarization values))/(FAM is glimmering for positive control standard items
Light polarization value-negative control standard items FAM fluorescence polarization values) } ÷ { (the R110 fluorescence polarization values-(positive controls of testing sample
Standard items R110 fluorescence polarization values-negative control standard items R110 fluorescence polarization values))/(positive control standard items R110 fluorescence is inclined
Shake value-negative control standard items R110 fluorescence polarization values) }
Embodiment：
Detect following each exons mutation index of cell sample EGFR allele 19 to be measured and to EGFR targeted inhibition agent
Sensitiveness.
The present embodiment agents useful for same includes：Amplified reaction is carried out in 25uL systems, adds 10 × PCR reaction buffers
2.5 μ L, concentration is dGTP, dCTP, dATP and dTTP of 2.0mmol/L, archaeal dna polymerase 1-2 units, 2.5mM MgCL2。
The forward and reverse primer of EGFR19 extrons of 0.5uM, the EGFR19 extron reverse primers of 0.07uM show outside the EGFR19 of 0.5uM
Sub- R110 marks wild-type probe, EGFR19 extrons FAM mark saltant type probes, template 5uL.
Add negative control standard items 5uL (pGEM-T-easy carriers 10000 copy/ml) in negative control reaction, it is positive right
According in reaction plus positive control standard items 5uL (the extron wild gene product pGEM-T-wild-19 of allele containing EGFR 19
1000 copy/ml of the clone and Varia nt in exon gene outcome pGEM-T-delmutation-19 of allele containing EGFR 19 clones
1000 copies/ml).1/1 cell mixing sample to be measured：By human lung carcinoma cell H292 (the extron wild type of EGFR allele 19,
It is insensitive to EGFR targeted inhibition agent) and HCC827 (the extron E746-A750del of EGFR allele 19 mutation, to EGFR targets
It is sensitive to inhibitor) mix in 1/1 ratio, DNA is extracted, 5uL adds amplified reaction.2/1 cell mixing sample to be measured：By people's lung
Cancer cell H292 (the extron wild type of EGFR allele 19, insensitive to EGFR targeted inhibition agent) and HCC827 (EGFR etc.
The position extron E746-A750del mutation of gene 19, sensitive to the agent of EGFR targeted inhibitions) mix in 2/1 ratio, extract DNA, 5uL
Add amplified reaction.
Reagent is expanded, amplification reaction condition is：After 94 DEG C of denaturation 5min, 95 DEG C are incubated 10 seconds, and 55 DEG C are incubated 10
Second, 72 DEG C are incubated 20 seconds, 45 circulations, and last circulation is without 72 DEG C of incubations.Detection amplified reaction dead end product FAM, R110
Fluorescence polarization value FP.
Testing result：
| Template/fluorescence polarization value (mp) | FAM(mp) | R110(mp) |
| Negative control | 41 | 38 |
| Positive control | 82 | 76 |
| 1/1 cell mixing sample to be measured | 126 | 115 |
| 2/1 cell mixing sample to be measured | 125 | 191 |
Height result of variations according to above FAM and R110 fluorescence polarization value and than being worth to cell mixing sample to be measured
The exons mutation index of EGFR allele 19：
The exons mutation indexes of 1/1 cell mixing sample EGFR 19 to be measured={ (126- (82-41))/(82-41) } ÷
{〔115-(76-38)〕/(76-38)}≈1
The exons mutation indexes of 2/1 cell mixing sample EGFR 19 to be measured={ (125- (82-41))/(82-41) } ÷
{〔191-(78-38)〕/(78-38)}≈0.5
I.e.：The exons mutation index of EGFR allele 19 of 1/1 cell mixing sample to be measured is about 1,2/1 mixing to be measured
The exons mutation index of cell sample EGFR allele 19 is about 0.5；Mutation index person high is quicker to EGFR targeted inhibition agent
Sense, i.e. 1/1 cell mixing sample is more sensitive to EGFR targeted inhibition agent compared with 2/1 cell mixing sample.
Present disclosure is not limited to cited by embodiment, and those of ordinary skill in the art are by reading description of the invention
And any equivalent conversion taken technical solution of the present invention, it is claim of the invention and is covered.
SEQUENCE LISTING
<110>The Fourth Military Medical University of P.L.A
<120>The method of the homogeneous exons mutation index of detection EGFR allele 19
<130> 201701
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213>Artificial sequence
<400> 1
gtgcatcgct ggtaacatcc 20
<210> 2
<211> 21
<212> DNA
<213>Artificial sequence
<400> 2
gaagcaacat ctccgaaaag c 21
<210> 3
<211> 20
<212> DNA
<213>Artificial sequence
<400> 3
gagggaatta agagaagctc 20
<210> 4
<211> 17
<212> DNA
<213>Artificial sequence
<400> 4
gccaaaacat ctccggc 17
Claims (6)
1. the method for homogeneously detecting the exons mutation index of EGFR allele 19, it is characterised in that：
Agents useful for same includes：
MgCl2, 10 × PCR reaction buffers, the extron DNA primer of dNTPs, EGFR allele 19, outside EGFR allele 19
Show sub- E746-A750, outside the FAM label probes of the 15bp deletion mutations DNA of 2236 to 2250 bit bases, EGFR allele 19
Show sub- E746-A750, the R110 label probes of the 15bp wild types DNA of 2236 to 2250 bit bases, template：Detected sample
DNA, negative control standard items are wild pGEM-T-easy empty carriers, positive control the standard items i.e. extron of EGFR allele 19
The mixture of raw type reference standards and 19 exons mutation type reference standards, concentration is 1000 copies/ml；
Reagent is expanded respectively, the detection of FAM and R110 fluorescence polarization values is then carried out respectively to amplified production, it is positive right
More than negative control standard items FAM and R110 fluorescence polarization value 30mp is respectively greater than according to standard items FAM and R110 fluorescence polarization value
Then to expand successfully, the difference height according to detected sample FAM and R110 fluorescence polarization value and positive and negative reference standards becomes
Change the exons mutation index of EGFR allele 19 with ratio interpretation detected sample.
2. the method for the homogeneous exons mutation index of detection EGFR allele 19 according to claim 1, its feature exists
In：
Amplified reaction is carried out in 25uL systems, adds the μ L of 10 × PCR reaction buffers 2.5, and concentration is 2.0mmol/L's
DGTP, dCTP, dATP and dTTP, archaeal dna polymerase 1-2 units, concentration is 1.5-3.0mM MgCL2.Concentration is 0.5uM's
The extron sense primer of EGFR gene 19 and 3 ' 2 kind probes of the end with fluorescence labeling, the EGFR equipotentials of concentration 0.1uM-0.05uM
The extron anti-sense primer of gene 19；5 μ L template DNAs；
PCR buffer solutions are 1.0U Taq polymerase.
3. the method for the homogeneous exons mutation index of detection EGFR allele 19 according to claim 1, its feature exists
In：
According to EGFR allele GenBank sequences, the primer of the extron E746-A750 of allele containing EGFR 19 is covered in design synthesis
And 19 extron wild types, saltant type probe：
EGFR19 forward primers 5'-gtgcatcgctggtaacatcc-3'；
EGFR19 reverse primers 5'-gaagcaacatctccgaaaagc-3'；
19 extron wild-type probes：5’-gagggaattaagagaagctc–R110 3’；
19 exons mutation type probes：5’-gccaaaacatctccggc-FAM 3’.
4. the method for the homogeneous exons mutation index of detection EGFR allele 19 according to claim 1, its feature exists
In：
Amplification reaction condition is：After 94 DEG C of denaturation 5min, 95 DEG C are incubated 1 second, and 60 DEG C are incubated 1 second, and 72 DEG C are incubated 10 seconds, and 45 are followed
Ring, last circulation is without 72 DEG C of incubations.
5. the method for the homogeneous exons mutation index of detection EGFR allele 19 according to claim 1, its feature exists
In：
The extron negative control standard items template of amplification EGFR allele 19 is pGEM-T-easy empty carriers, positive right respectively
It is that the extron wild type control standard items 1000 of EGFR allele 19 copy/ml+19 exons mutation types according to standard items template
Reference standards 1000 copy/ml and DNA sample to be measured, the extron genes of interest amplification of EGFR allele 19 is obtained respectively
Product.
6. the method for the homogeneous exons mutation index of detection EGFR allele 19 according to claim 1, its feature exists
In：
Methods described detects that negative control standard items, positive control standard items and testing sample EGFR allele 19 are outer aobvious respectively
FAM the and R110 fluorescence polarization values of the final amplified production of sub- genes of interest；
Positive control standard items R110, FAM fluorescence polarization value is respectively greater than negative control standard items R110, FAM fluorescence polarization value
More than 30mp is then to expand successfully；
The exons mutation index of EGFR allele 19 of testing sample={ (the FAM fluorescence polarization values of testing sample-(positive right
According to standard items FAM fluorescence polarization values-negative control standard items FAM fluorescence polarization values))/(positive control standard items FAM fluorescence is inclined
Shake value-negative control standard items FAM fluorescence polarization values) } ÷ { (the R110 fluorescence polarization values of testing sample-(positive control standard
Product R110 fluorescence polarization values-negative control standard items R110 fluorescence polarization values))/(positive control standard items R110 fluorescence polarizations
Value-negative control standard items R110 fluorescence polarization values) }.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050130197A1 (en) * | 2003-09-19 | 2005-06-16 | Do Ernest U. | Homogeneous fluorescence polarization assay for high throughput screening |
| CN102676661B (en) * | 2012-04-27 | 2014-12-31 | 中国人民解放军第四军医大学 | Fluorescent polarization based homogeneous phase detection method of single nucleotide polymorphism of codon118 of ERCC1 (excision repair cross-complementing 1) gene |
-
2017
- 2017-01-19 CN CN201710044212.3A patent/CN106834458A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050130197A1 (en) * | 2003-09-19 | 2005-06-16 | Do Ernest U. | Homogeneous fluorescence polarization assay for high throughput screening |
| CN102676661B (en) * | 2012-04-27 | 2014-12-31 | 中国人民解放军第四军医大学 | Fluorescent polarization based homogeneous phase detection method of single nucleotide polymorphism of codon118 of ERCC1 (excision repair cross-complementing 1) gene |
Non-Patent Citations (4)
| Title |
|---|
| WENCHAO LIU等: "Detection of EGFR Mutation in Tissue Samples of Non-small-cell Lung Cancer by a Fluorescence Polarization Assay", 《DIAGN MOL PATHOL》 * |
| ZHANG WEI等: "EGFR Promoter Methylation, EGFR Mutation, and HPV Infection in Chinese Cervical Squamous Cell Carcinoma", 《APPLIED IMMUNOHISTOCHEMISTRY & MOLECULAR MORPHOLOGY》 * |
| 李伟等: "《分子诊断学》", 30 September 2015 * |
| 杨阳等: "肺癌中EGFR外显子突变丰度的荧光偏振检测", 《现代肿瘤医学》 * |
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