CN102168129A - Method for detecting nasopharyngeal carcinoma radiosensitive genes and primers and fluorescence probes thereof - Google Patents
Method for detecting nasopharyngeal carcinoma radiosensitive genes and primers and fluorescence probes thereof Download PDFInfo
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Abstract
The present invention discloses a method for detecting nasopharyngeal carcinoma radiosensitive genes and primers and fluorescence probes thereof. The primers used for detecting nasopharyngeal carcinoma radiosensitive genes are composed f 18 DNA sequences ranging from SEQ ID:1 to SEQ ID:18 in a sequence table. The fluorescence probes are composed of 9 DNA sequences ranging from SEQ ID:19 to SEQ ID:27 in the sequence table, wherein the 5' end of the DNA sequences is labeled with report fluorescence groups with different luminescence colors, and the 3' end is labeled with nonluminous quenching groups. The method provides a new way for detecting nasopharyngeal carcinoma radiosensitivity and serves as a guide in curing nasopharyngeal carcinoma.
Description
Technical field
The invention belongs to biological technical field, obtain cDNA by reverse transcription (RT) mRNA sample earlier, utilize the real-time fluorescence quantitative PCR technology again, gene expression amount in the detected sample of accurate quantification.
Background technology
(its cause of disease relates to interaction and the environment and the dietary factor of Epstein-Barr virus, inherited genetic factors (genetic predisposition), multiple oncogene and cancer suppressor gene to nasopharyngeal carcinoma for Nasopharyngeal carcinoma, NPC) good sending out in southern china each province and south east asia.Along with the develop rapidly of molecular biology and technology thereof, people recognize that by the further investigation to oncogene and cancer suppressor gene canceration is a complex process that relates to multiple gene event in recent years.The research of nasopharyngeal carcinoma generation, development and biological behaviour thereof has also entered gene level, and is in the ascendant about the research of nasopharyngeal carcinoma oncogene and cancer suppressor gene.Nasopharyngeal carcinoma has a lot of differentiation types, is common in pharyngeal recess, is positioned at the back inner side-wall of pharyngeal opening of auditory tube medial angle.Between 40～60 years old, the male sex is more than the women mostly for the nasopharyngeal carcinoma age of onset.The most of regional nasopharyngeal carcinoma sickness rate in the world is lower, generally 1/10
5Below.The morbidity of nasopharyngeal carcinoma has tangible regional aggregation and ethnicity phenomenon, is more common in the yellow, and the patient mainly concentrates on Guangdong, Chaozhou-Shantou region and Foochow dialect crowd and these crowds migrate extra large epigenesist.China south (provinces such as Guangdong, Guangxi, Fujian and Hunan) and some countries of South East Asia are the highest areas of nasopharyngeal carcinoma sickness rate, the whole world, and particularly midwestern Zhaoqing, Guangdong, Foshan and area, Guangzhou are higher.The male sex NPC sickness rate of Zhaoqing four meetings is 25.12/10
5, the women is 12.11/10
5Live in the middle part, Guangdong Province and say that the local male sex's sickness rate of speaking in Guangdong is up to 30/10
5～50/10
5
Epidemiological survey points out that the cause of disease of nasopharyngeal carcinoma may be relevant with following factors: 1. ebv infection.2. environment and diet: environmental factors also is a kind of reason of bringing out nasopharyngeal carcinoma.In Guangdong, the rice and the low Qu Weigao of sending out of the microelement nickel content in the water of cancer district occurred frequently found in investigation.In nasopharyngeal carcinoma patient's hair, nickel content is also high.Animal experiment shows that nickel can promote nitrosamine to bring out nasopharyngeal carcinoma.The edible cured fish of report also being arranged and pickle food is southern china nasopharyngeal carcinoma high risk factor, and with age of food cured fish, the time limit of use, amount and cooking method are relevant.3. inherited genetic factors: nasopharyngeal cancer patient has race and family's clustering phenomena, as the southern china people offspring who lives in other countries still keeping very high nasopharyngeal carcinoma sickness rate, and this prompting nasopharyngeal carcinoma may be a heredopathia.
Because the region of anatomy of nasopharynx exists deeply, the anatomical structure complexity, pathology is mainly non-keratinization type undifferentiated carcinoma and is prone to cervical lymph node and distant metastasis, and in the patient of first visit, III, IV phase nasopharyngeal carcinoma account for about 70% of total case.A very long time all is the standard care means of nasopharyngeal carcinoma to radiotherapy in the past, but 5 years survival rates of radiotherapy alone nasopharyngeal carcinoma only about 50%, and 5 years survival rates of advanced NPC (III, IV) are lower, about 10%～40%, the major cause of treatment failure is local recurrence and distant metastasis.Therefore, study result of treatment how to improve the nasopharyngeal carcinoma patient and become very necessary and meaningful.Because of nasopharyngeal carcinoma is the tumour responsive relatively to chemotherapy, therefore seek the chemotherapeutics of effective better tolerance and the combination of chemicotherapy is a hot research in recent years.In addition,, found that some biological indicators can predict the prognosis and the curative effect of malignant tumour, played a significant role in risk assessment and when formulating therapeutic strategy along with development of molecular biology.
Studies show that ZNF187, CLSTN1, HBG2, PRO0149, NME6, SMARCC1, GL009, GL010, there were significant differences in the expression of these 9 genes of GL01 in nasopharyngeal carcinoma radiation-sensitive type patient and nasopharyngeal carcinoma radiation resistance type patient, can provide guidance for the treatment of nasopharyngeal carcinoma by detecting these 9 expression of gene situations.
Traditional gene diagnosis technology is mainly used the method for polymerase chain reaction (PCR) or RT-polymerase chain reaction (RT-PCR) testing gene that increases in a large number, then by ethidium bromide staining and the quantitative testing gene of agarose gel electrophoresis.Above method has high highly sensitive, but the characteristics of PCR-based reaction, the product amount of reaction end often can't correctly reflect the content that reacts starting stage DNA, therefore quantitatively accurate inadequately, the resolving power of its detection simultaneously is not high yet, if the difference of the amount of DNA is lower than five times, often can not detect.Continuous perfect real-time quantitative PCR technology has added fluorophor in the PCR reaction system in recent years, utilizes the accumulation of fluorescent signal to monitor the process of whole PCR, at last by Ct value and typical curve to the DNA accurate quantification in the sample.Because a product to the exponential phase reaction detects the signal amplification with fluorophor, it has overcome the quantitative out of true of conventional P CR, the shortcoming that resolving power is low.
Summary of the invention
The purpose of this invention is to provide primer at the qualitative and quantitative analysis of nasopharyngeal carcinoma radiosensitivity gene.
For solving the problems of the technologies described above, the present invention takes following technical scheme:
Be used to detect the primer of ZNF187 gene, right by the primer that SEQ ID NO:1 in the sequence table and SEQ ID NO:2 form.
Be used to detect the primer of CLSTN1 gene, right by the primer that SEQ ID NO:3 in the sequence table and SEQ ID NO:4 form.
Be used to detect the primer of HBG2 gene, right by the primer that SEQ ID NO:5 in the sequence table and SEQ ID NO:6 form.
Be used to detect the primer of PRO0149 gene, right by the primer that SEQ ID NO:7 in the sequence table and SEQ ID NO:8 form.
Be used to detect the primer of NME6 gene, right by the primer that SEQ ID NO:9 in the sequence table and SEQ ID NO:10 form.
Be used to detect the primer of SMARCC1 gene, right by the primer that SEQ ID NO:11 in the sequence table and SEQ ID NO:12 form.
Be used to detect the primer of GL009 gene, right by the primer that SEQ ID NO:13 in the sequence table and SEQ ID NO:14 form.
Be used to detect the primer of GL010 gene, right by the primer that SEQ ID NO:15 in the sequence table and SEQ ID NO:16 form.
Be used to detect the primer of GL011 gene, right by the primer that SEQ ID NO:17 in the sequence table and SEQ ID NO:18 form.
The present invention also provides the fluorescent probe that carries out detection by quantitative at nasopharyngeal carcinoma radiosensitivity gene.
For solving the problems of the technologies described above, the present invention takes following technical scheme:
Being used to detect the fluorescent probe of ZNF187 gene, is the dna sequence dna by SEQ ID NO:19 in the sequence table and SEQ IDNO:20.
Being used to detect the fluorescent probe of CLSTN1 gene, is the dna sequence dna by SEQ ID NO:21 in the sequence table and SEQ IDNO:22.
Being used to detect the fluorescent probe of HBG2 gene, is the dna sequence dna by SEQ ID NO:23 in the sequence table and SEQ ID NO:24.
Being used to detect the fluorescent probe of PRO0149 gene, is the dna sequence dna by SEQ ID NO:25 in the sequence table and SEQ IDNO:26.
Being used to detect the fluorescent probe of NME6 gene, is the dna sequence dna by SEQ ID NO:27 in the sequence table and SEQ ID NO:28.
Being used to detect the fluorescent probe of SMARCC1 gene, is the dna sequence dna by SEQ ID NO:29 in the sequence table and SEQ IDNO:30.
Being used to detect the fluorescent probe of GL009 gene, is the dna sequence dna by SEQ ID NO:31 in the sequence table and SEQ ID NO:32.
Being used to detect the fluorescent probe of GL010 gene, is the dna sequence dna by SEQ ID NO:33 in the sequence table and SEQ ID NO:34.
Being used to detect the fluorescent probe of GL011 gene, is the dna sequence dna by SEQ ID NO:35 in the sequence table and SEQ ID NO:36.
Described probe is through fluorescent mark, and 5 ' end is marked with the different report fluorophor of glow color, and 3 ' end is marked with non-luminous cancellation group.The report fluorophor is FAM or VIC, and non-luminous quenching group is NFQ.
The present invention also provides a kind of test kit that detects nasopharyngeal carcinoma radiosensitivity gene, can comprise above-mentioned primer and fluorescent probe.
Embodiment
The detection by quantitative of ZNF187 genetic expression
1. the extracting of the total RNA of sample and quality inspection
Collect case sample 20 examples, CT examination result during according to pharynx nasalis irradiation 60Gy, tumor regression＜40% is resisted for radiation,＞80% be the evaluation index of radiation-sensitive as radiosensitivity, be divided into the radiation-sensitive group and radiate the opposing group.20 is routine except that an example classification is not clear, responsive 8 examples of organizing, and 11 examples are organized in opposing.
The case sample is at the nasopharyngeal fiberscope direct vision biopsy, and biopsy is transferred in the stone roller alms bowl with the liquid nitrogen precooling, with the pestle tissue of milling, constantly adds liquid nitrogen therebetween, and is Powdered until being milled into.
To be milled into pulverous sample, be transferred in the homogenate pipe that adds an amount of TRIzol reagent in advance, the homogenate pipe will be placed ice bath, on the tissue homogenate pulverizer, carry out homogenate.Homogenate is not glued and is not had particle to homogenate and gets final product.Annotate: put into homogenate Guan Qianyong scales/electronic balance weighing and record,, be generally less than tissue adding 5 or the 10ml lysate of 1g according to the ratio adding TRIzol reagent of weight in 1: 2.
Homogenate is transferred in the 15ml centrifuge tube, in-4 ℃, 12000rpm, centrifugal 10min.
The careful supernatant liquor of drawing moves into new 15ml centrifuge tube, places 5min at 15～30 ℃.
In homogenate, add chloroform, cover tight centrifuge tube lid, use the forced oscillation centrifuge tube, place 3min at 15～30 ℃.
Annotate: the ratio of chloroform and homogenate is 1: 5.In-4 ℃, 12000rpm, centrifugal 15min.
The careful centrifuge tube that takes out is drawn supernatant to another 15ml centrifuge tube from whizzer.Annotate: must be careful when drawing supernatant, it is light that action is wanted, and can not make that the intermediary protein ingredient is floating to come up, otherwise directly have influence on the quality of RNA.
Add Virahol, put upside down centrifuge tube gently, place 10min at 15～30 ℃ with abundant mixing liquid.Annotate: the amount that adds Virahol is isopyknic supernatant liquor.In-4 ℃, 12000rpm, centrifugal 10min.
Supernatant discarded adds 75% ethanol 1ml along tube wall slowly, breaks repeatedly with the rifle head and beats, and the material of the attached wall of white is all melted, and move in the 1.5ml centrifuge tube.Of short duration vortex on vortice, in-4 ℃, 8000rpm, centrifugal 10min.
Carefully abandon supernatant, of short duration centrifugal, liquid-transfering gun is drawn all supernatants (i.e. 75% ethanol), drying precipitated 5min in super quiet worktable.
The complete RNA of Milli-Q water that adds Rnase-free melts post precipitation ,-80 ℃ of preservations.The Milli-Q water that adds 40-50U.
Ultraviolet spectrophotometer is measured the purity of total RNA: get 1 μ L solution and be dissolved among the 99 μ L Tris, and detect OD260 with ultraviolet spectrophotometer, OD280, OD320, the absorption value of OD230.Annotate: the Ratio value is that OD260/OD280 〉=1.800 are qualified for quality inspection.
Total thermally-stabilised experiment of RNA: get in the 0.5ml Eppendorf pipe and add RNA 400ng respectively, get two pipes and handled 1 hour for 70 ℃, another pipe-20 ℃ was handled 1 hour, carried out the agarose electrophoresis of 1.5%Rnase-free simultaneously, observed the quality of RNA.Annotate: show that the RNA band is clear in 70 ℃ and-20 ℃ of two experiments, the border is fuzzy, and prompting RNA does not degrade, and quality inspection is qualified.
2. reverse transcription PCR
(1) in 500 μ l Eppendorf tubes, add total RNA 1-5 μ g, replenish an amount of DEPC H2O and make cumulative volume reach 11 μ l.In pipe, add 10 μ M Oligo (dT) 12-18,1 μ l, gently mixing, centrifugal.
(2) 70 ℃ of heating 10min insert Eppendorf tube in the ice bath 1min at least immediately.
The mixture that adds following reagent then:
10×PCR?buffer 2μl
25mM?MgCl2 2μl
10mM?dNTPmix 1μl
0.1M?DTT 2μl
Mixing is centrifugal gently.Hatch 2-5min for 42 ℃.
(3) add Superscript II 1 μ l, in 42 ℃ of water-baths, hatch 50min.
(4) heat 15min with termination reaction in 70 ℃.
(5) will manage in the insertion ice, add RNase H 1 μ l, hatch 20min for 37 ℃, the RNA of degrade residual.-20 ℃ of preservations are standby.
3. augmentation detection: carry out on 7900 quantitative real time PCR Instruments, total reaction volume is 25 μ l, 12.5 μ l PCR reaction mixtures wherein, and 2.5 μ l detect with primer, fluorescent probe, 5 μ l testing sample cDNA.Reaction conditions: 95 ℃ of sex change in 10 minutes, carry out subsequently 95 ℃ 15 seconds, 60 ℃ of 45 circulations in a minute place 4 ℃ at last.According to typical curve, calculate the copy number of ZNF187 gene in each testing sample.
4. result
Sequence table
<160>64
<210>1
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>1
agtgtgaagt?tacaaagcct?ga
<210>2
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>2
ttatctatgt?atccattgtg?ga
<210>3
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>3
agccacatag?ggaaaggctg?cg
<210>4
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>4
gcccttctct?gtgactgagg?at
<210>5
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>5
atttcacaga?ggaggacaag?gc
<210>6
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>6
atctattct?gctaagagat?ca
<210>7
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>7
cccttggagc?ggccacgcct?ca
<210>8
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>8
gaggccgcgg?gaggccgcgg?ag
<210>9
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>9
aagcaacaag?ttcctgattg?ta
<210>10
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>10
ggcctagctc?accacagcac?at
<210>11
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>11
tcttggggct?gcttacaagt?at
<210>12
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>12
agcgcagaag?tcagaaagag?ga
<210>13
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>13
acccgtattt?gctgagcatt?tt
<210>14
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>14
ttacccctta?gccattgaga?ct
<210>15
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>15
tccctaatcc?tctgcaattc?ct
<210>16
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>16
gaagtacttt?tgaagactcc?ca
<210>17
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>17
tgtcatggag?atgtttgtat?ac
<210>18
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>18
tttcccactt?tttcttgaaa?ga
<210>19
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>19
tgggccattc?ggcaggaaga?agga
<210>20
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>20
tgggccattc?ggcaggaaga?agga
<210>21
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>21
acttccttgg?gagatgccat?aaag
<210>22
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>22
gcggcggcgg?cggcagtggg?gagg
<210>23
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>23
cccgcaacac?cacccatggt?tcgg
<210>24
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>24
catattcttc?ctcacaagac?gatg
<210>25
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>25
cataaatgaa?gcctctttgg?ggtg
<210>26
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>26
agaaatgtac?aagctgtatc?ctaa
<210>27
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>27
ggagctttcc?ctgtctgggt?ttag
Sequence table
<160>64
<210>1
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>1
agtgtgaagt?tacaaagcct?ga
<210>2
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>2
ttatctatgt?atccattgtg?ga
<210>3
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>3
agccacatag?ggaaaggctg?cg
<210>4
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>4
gcccttctct?gtgactgagg?at
<210>5
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>5
atttcacaga?ggaggacaag?gc
<210>6
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>6
atctattct?gctaagagat?ca
<210>7
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>7
cccttggagc?ggccacgcct ca
<210>8
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>8
gaggccgcgg?gaggccgcgg?ag
<210>9
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>9
aagcaacaag?ttcctgattg?ta
<210>10
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>10
ggcctagctc?accacagcac?at
<210>11
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>11
tcttggggct?gcttacaagt?at
<210>12
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>12
agcgcagaag?tcagaaagag?ga
<210>13
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>13
acccgtattt?gctgagcatt?tt
<210>14
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>14
ttacccctta?gccattgaga?ct
<210>15
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>15
tccctaatcc?tctgcaattc?ct
<210>16
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>16
gaagtacttt?tgaagactcc?ca
<210>17
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>17
tgtcatggag?atgtttgtat?ac
<210>18
<211>22
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>18
tttcccactt?tttcttgaaa?ga
<210>19
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>19
tgggccattc?ggcaggaaga?agga
<210>20
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>20
tgggccattc?ggcaggaaga?agga
<210>21
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>21
acttccttgg?gagatgccat?aaag
<210>22
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>22
gcggcggcgg?cggcagtggg?gagg
<210>23
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>23
cccgcaacac?cacccatggt?tcgg
<210>24
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>24
catattcttc?ctcacaagac?gatg
<210>25
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>25
cataaatgaa?gcctctttgg?ggtg
<210>26
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>26
agaaatgtac?aagctgtatc?ctaa
<210>27
<211>24
<212>DNA
＜213〉artificial sequence
<220>
<230>
<400>27
ggagctttcc?ctgtctgggt?ttag
Claims (21)
1. be used to detect the primer of ZNF187 gene, right by the primer that SEQ ID NO:1 in the sequence table and SEQ ID NO:2 form.
2. be used to detect the primer of CLSTN1 gene, right by the primer that SEQ ID NO:3 in the sequence table and SEQ ID NO:4 form.
3. be used to detect the primer of HBG2 gene, right by the primer that SEQ ID NO:5 in the sequence table and SEQ ID NO:6 form.
4. be used to detect the primer of PRO0149 gene, right by the primer that SEQ ID NO:7 in the sequence table and SEQ ID NO:8 form.
5. be used to detect the primer of NME6 gene, right by the primer that SEQ ID NO:9 in the sequence table and SEQ ID NO:10 form.
6. be used to detect the primer of SMARCC1 gene, right by the primer that SEQ ID NO:11 in the sequence table and SEQ ID NO:12 form.
7. be used to detect the primer of GL009 gene, right by the primer that SEQ ID NO:13 in the sequence table and SEQ ID NO:14 form.
8. be used to detect the primer of GL010 gene, right by the primer that SEQ ID NO:15 in the sequence table and SEQ ID NO:16 form.
9. be used to detect the primer of GL011 gene, right by the primer that SEQ ID NO:17 in the sequence table and SEQ ID NO:18 form.
10. being used to detect the fluorescent probe of ZNF187 gene, is the dna sequence dna of SEQ ID NO:19 in the sequence table.
11. be used to detect the fluorescent probe of CLSTN1 gene, be the dna sequence dna of SEQ ID NO:20 in the sequence table.
12. be used to detect the fluorescent probe of HBG2 gene, be the dna sequence dna of SEQ ID NO:21 in the sequence table.
13. be used to detect the fluorescent probe of PRO0149 gene, be the dna sequence dna of SEQ ID NO:22 in the sequence table.
14. be used to detect the fluorescent probe of NME6 gene, be the dna sequence dna of SEQ ID NO:23 in the sequence table.
15. be used to detect the fluorescent probe of SMARCC1 gene, be the dna sequence dna of SEQ ID NO:24 in the sequence table.
16. be used to detect the fluorescent probe of GL009 gene, be the dna sequence dna of SEQ ID NO:25 in the sequence table.
17. be used to detect the fluorescent probe of GL010 gene, be the dna sequence dna of SEQ ID NO:26 in the sequence table.
18. be used to detect the fluorescent probe of GL011 gene, be the dna sequence dna of SEQ ID NO:27 in the sequence table.
19. according to the described dna sequence dna of claim 10-18, it is characterized in that: 5 ' end is marked with the different report fluorophor of glow color, and 3 ' end is marked with non-luminous cancellation group.
20. according to the described fluorescent probe of claim 10-18, it is characterized in that: described report fluorophor is FAM or VIC, and non-luminous quenching group is NFQ.
21. a test kit that detects nasopharyngeal carcinoma radiosensitivity gene comprises described primer of claim 1-9 and the described fluorescent probe of claim 10-18.
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| CN2010101145368A CN102168129A (en) | 2010-02-26 | 2010-02-26 | Method for detecting nasopharyngeal carcinoma radiosensitive genes and primers and fluorescence probes thereof |
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|---|---|---|---|
| CN2010101145368A CN102168129A (en) | 2010-02-26 | 2010-02-26 | Method for detecting nasopharyngeal carcinoma radiosensitive genes and primers and fluorescence probes thereof |
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| CN111304319B (en) * | 2020-04-10 | 2023-07-25 | 北京瑞康我科技有限公司 | Diagnostic marker for diseases and application thereof |
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