CN104593490A - Application of USP8 gene detection object in preparation for ACTH-type pituitary adenoma molecular pathological diagnosis and typing products - Google Patents
Application of USP8 gene detection object in preparation for ACTH-type pituitary adenoma molecular pathological diagnosis and typing products Download PDFInfo
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- CN104593490A CN104593490A CN201410831608.9A CN201410831608A CN104593490A CN 104593490 A CN104593490 A CN 104593490A CN 201410831608 A CN201410831608 A CN 201410831608A CN 104593490 A CN104593490 A CN 104593490A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention discloses an application of a USP8 gene detection object in preparation for ACTH-type pituitary adenoma molecular pathological diagnosis and typing products. The invention provides a kit used for detecting the ACTH-type pituitary adenoma subtypes of a to-be-detected patient with the ACTH-type pituitary adenoma, wherein the kit comprises a comparison card for recording a detection standard and a sequencer. Experiments of the application disclosed by the invention prove that USP8 gene mutation is peculiar in the ACTH-type pituitary adenoma, and does not exist in other types of pituitary adenoma, so that the USP8 gene mutation can be used as a specific molecular pathological diagnosis marker for the ACTH-type pituitary adenoma. The ACTH-type pituitary adenoma can be further divided into two subtypes, namely a mutant type (with the USP8 gene mutation) and a wide type (without the USP8 gene mutation), so that an individualized clinical treatment strategy is adopted in time.
Description
Technical field
The present invention relates to biological technical field, particularly relate to the application of USP8 gene test thing in preparation ACTH type pituitary adenoma molecular diagnosis and somatotype product.
Background technology
Pituitary tumor (Pituitary adenomas) is the modal neuroendocrine of human body, is also one of modal cerebral tumor (sickness rate accounts for the 2nd).By the internal secretion characteristic that they are different, lactotropin type, tethelin type, thyroliberin type can be divided into, thyrotropic hormone type, without some hypotypes such as hormone-type and mixed type.Tumour by the various hormone of abnormal secretion, causes patient to occur the significant endocrine regulation symptom such as infertile, acromegaly, Cushing syndromes, hyperthyroidism.Pituitary tumor incidence of occult, poor growth, when patient occurs that clinical symptom is gone to medical, tumour often bulky, in invasive growth, the contiguous important anatomy structure of compressing, causes patient to occur the neurological deficit symptoms such as the decline of primary pituitary hypofunction, binocular vision and parmlysis of cranial nerve, have a strong impact on the quality of life of patient, even threat to life.
A large amount of ACTH secretes by the corticotroph in pituitary gland in thyroliberin (ACTH) type pituitary adenoma system, promote the hydrocortisone that acth secretion is a large amount of, finally cause endogenous blood hypercortisolism, then a series of metabolism disorder and pathological change is produced, spy is referred to as hypercortisolism (Cushing ' s diease), and quantity accounts for 85% of ACTH dependency hypercortisolism.Hypercortisolism is a kind of serious endocrinopathy, few spontaneous remission, if diagnosis and treatment not in time, case fatality rate is very high.This patient's main manifestations is central obesity, hypertension, cardiovascular disorder, metabolism disorder syndrome, osteoporosis etc.Although the Molecular pathogenesis of hypercortisolism is widely studied, it is still indefinite that ACTH produces superfluous pathogenic genetic mechanism.The clinical treatment of current pituitary tumor is based on excision, and medicine and stereotactic radiotherapy are auxiliary, but only has small number of patients to obtain healing by these methods! Most of through cure the disease example there is tumor recurrence at shorter remission after date.Therefore bite clinically and treat new methods for the treatment of.In addition, find that ACTH type pituitary adenoma can present distinct Radiologic imaging and clinical phenotypes: Partial tumors volume is small, and focus is the distribution of disperse shape, operation consent location is very difficult; And Partial tumors bulky, simultaneously to periphery invasive growth, extremely difficult acquisition tumor resection in operation.Infer that they may belong to the different hypotype of ACTH type pituitary tumor two kinds, different clinical management strategy should be taked to it.There is no specific molecular diagnostic markers thing at present and they can be carried out differential diagnosis.
Summary of the invention
An object of the present invention is to provide the test kit for detecting ACTH type pituitary adenoma patients tumors subtypes to be measured.
Test kit provided by the invention, comprises the comparison card and sequenator of recording examination criteria;
Described examination criteria is as follows:
If in ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then ACTH type pituitary adenoma patients tumour to be measured is or candidate is ACTH type pituitary adenoma saltant type; If in ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with described USP8 gene full wild-type sequence or corresponding part fragment sequence, then ACTH type pituitary adenoma patients tumour to be measured is or candidate is ACTH type pituitary adenoma wild-type;
Described USP8 gene wild-type sequence is sequence 1 in sequence table.
Mentioned reagent box also comprises the primer set for increase USP8 full length gene or its Partial Fragment;
Described primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Above-mentioned primer or primer pair or the wherein equal independent packaging of each bar primer, each primer pair is used alone.
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
Another object of the present invention is to provide a kind of for helping the primer set detecting ACTH type pituitary adenoma patients tumors subtypes to be measured.
Primer set provided by the invention, is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
Mentioned reagent box or the application of above-mentioned primer set in the product preparing detection or auxiliary detection ACTH type to be measured pituitary adenoma patients tumors subtypes are also the scope of protection of the invention.
Whether the 3rd object of the present invention is to provide for auxiliary detection pituitary adenoma patients tumour to be measured is the test kit of ACTH type.
Test kit provided by the invention, comprises the comparison card and sequenator of recording examination criteria;
Described examination criteria is as follows:
If in pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then pituitary adenoma patients tumor candidate to be measured is ACTH type pituitary adenoma; If in pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with described USP8 gene full wild-type sequence or corresponding part fragment sequence, then pituitary adenoma patients tumor candidate to be measured is non-ACTH type pituitary adenoma;
Described USP8 gene wild-type sequence is sequence 1 in sequence table.
Mentioned reagent box also comprises the primer set for increase USP8 full length gene or its Partial Fragment;
Described primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
The present invention's the 4th object is to provide the primer set whether a kind of auxiliary detection pituitary adenoma patients tumour to be measured is ACTH type.
Whether auxiliary detection provided by the invention pituitary adenoma patients tumour to be measured is the primer set of ACTH type, is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
Whether mentioned reagent box or above-mentioned primer set are application in the product of ACTH type in preparation auxiliary detection pituitary adenoma patients tumour to be measured is also the scope of protection of the invention.
Two kinds of hypotypes of ACTH pituitary adenoma: it is wild hypotype that diameter of tumor is more than or equal to 1cm, diameter of tumor is less than 1cm for sudden change hypotype, and sudden change hypotype is that USP8 gene wild-type is undergone mutation the gene obtained, 17 kinds as shown in table 3 of concrete mutational formats.
Experiment of the present invention proves, USP8 transgenation by ACTH type pituitary adenoma peculiar, have no this transgenation in the pituitary adenoma of other types.ACTH pituitary adenoma can be further divided into two kinds of hypotypes, i.e. saltant type (USP8 transgenation occurs) and wild-type (USP8 gene is without sudden change), and the former average-volume is less than normal, and invasion and attack degree is little; Rear group of average-volume is bigger than normal, and invasion and attack degree is large.Whether can suddenly change and judge the different subtype of ACTH pituitary adenoma by detecting USP8 gene, whether can be also ACTH hypotype by USP8 gene auxiliary judgment pituitary adenoma of whether suddenling change, thus take different clinical management strategy in time.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Whether embodiment 1, USP8 gene suddenly change the application in assistant identification ACTH type pituitary adenoma hypotype
Carry out full exon to the tumour of 12 routine ACTH type pituitary adenomas and self blood sample DNA to check order comparative analysis (sequenator is Illumina Hiseq 2500), USP8 transgenation is there is in result in 8 routine tumours, gross tumor volume is little, in 4 routine tumours, USP8 gene is wild-type, and gross tumor volume is large; Therefore ACTH pituitary adenoma is divided into two kinds of hypotypes: diameter is more than or equal to for wild hypotype ACTH pituitary adenoma hypotype, diameter is less than for sudden change hypotype ACTH pituitary adenoma hypotype.
By 12 routine samples after false positive is got rid of in Sanger checking, in other 108 routine ACTH type pituitary adenomas, carry out USP8 gene type detect, the detection method of totally 120 routine samples is specific as follows:
First be designed for its primer pair of amplification according to USP8 gene (nucleotides sequence is classified as sequence 1 in sequence table), totally 3 pairs of primer pairs, often pair of primer pair all can increase USP8 Gene Partial fragment, and the concrete sequence of primer pair is as follows:
Primer pair 1:
1Primer Forward 5 '-ATTCACCCACCAACACTGTTCATA-3 ' (sequence 2)
1Primer Reverse 5 '-TTGTTTTCCCGATTAACTGTTGGA-3 ' (sequence 3)
Primer pair 2:
2Primer Forward 5 '-TCCATCTAAGTTTCTTGACCCAA-3 ' (sequence 4)
2Primer Reverse 5 '-TCCAACTCCCTGACACTAACA-3 ' (sequence 5)
Primer pair 3:
3Primer Forward 5 '-ATGTACCCACCGGAAATGGC-3 ' (sequence 6)
3Primer Reverse 5 '-CCTCAAAGGGAGATTCCGGG-3 ' (sequence 7)
With the genomic dna of above-mentioned 120 routine tumour patient tumours for template, carry out pcr amplification with above-mentioned primer pair 1, primer pair 2 and primer pair 3 respectively, obtain pcr amplification product.
PCR amplification system is as shown in table 1:
Table 1 is amplification system
| Component | Final concentration | Volume (μ L) |
| ddWater | 6.4 | |
| 2X Taq PCR Master Mix | 1X | 10 |
| Primer F | 400nM | 0.8 |
| Primer R | 400nM | 0.8 |
| DNA | 1ng/μL | 2 |
| Total | 20 |
Amplification program is as shown in table 2 below:
Table 2 is reaction conditions
The PCR primer of all primers obtained is checked order respectively, found that (the wild-type USP8 gene nucleotide series do not suddenlyd change is sequence 1 in sequence table) somatic variation and the ACTH type pituitary adenoma of ubiquitin-specific protease USP8 gene are closely related, concrete outcome is as follows:
In 120 routine tumours altogether, find that there is 75 routine tumours and there is USP8 transgenation (62.5%), have 17 kinds of mutation types, as shown in table 3: wherein 26 routine patients are shown in table 3 No. 1 mutational formats (wherein 1 routine patient is shown in table 3 No. 9 mutational formats simultaneously), 17 routine patients are shown in table 3 No. 2 mutational formats (wherein 1 routine patient is shown in table 3 No. 14 mutational formats simultaneously), 17 routine patients are shown in table 3 No. 3 mutational formats, 3 routine patients are shown in table 3 No. 4 mutational formats, 3 routine patients are shown in table 3 No. 5 mutational formats (wherein 1 routine patient is shown in table 3 No. 13 mutational formats simultaneously), 2 routine patients are shown in table 3 No. 6 mutational formats (wherein 1 routine patient is shown in table 3 No. 7 mutational formats simultaneously), 2 routine patients are shown in table 3 No. 7 mutational formats (wherein 1 routine patient is shown in table 3 No. 6 mutational formats simultaneously), 1 routine patient is shown in table 3 No. 8 mutational formats, 1 routine patient is shown in table 3 No. 10 mutational formats (wherein 1 routine patient is shown in table 3 No. 17 mutational formats simultaneously), 1 routine patient is shown in table 3 No. 11 mutational formats, 1 routine patient is shown in table 3 No. 12 mutational formats, 1 routine patient is shown in table 3 No. 15 mutational formats, 1 routine patient is shown in table 3 No. 16 mutational formats, in 75 routine patients, tumor average diameter is 0.84cm, and average-volume is 1.21cm
3, the ratio wherein in invasive growth tumour is 12.70%.
Altogether in 120 routine tumours, other 45 routine ACTH type pituitary adenoma patients USP8 genes are wild-type (nucleotides sequence is classified as sequence 1 in sequence table), and the mean diameter of this group tumour is 2.13cm, and average-volume is 6.6cm
3, the ratio wherein in invasive growth tumour is 41.3%.
Table 3 is USP8 gene 17 kind mutation type
Gene in above-mentioned table 3 is wild-type USP8 gene, and the sequence of its correspondence is sequence 1.
In the motif district that said gene sudden change is all gathered in 14-3-3 albumen and near, the interaction between USP8 and 14-3-3 albumen can be destroyed, remove ubiquitination pathway by what strengthen, prevent EGFR through lysosomal degradation.Along with the rising of EGFR content in cell, indirectly can promote that the generation of ACTH increases.
Therefore, the tumors subtypes of whether can be undergone mutation by USP8 gene detection or auxiliary detection ACTH type to be measured pituitary adenoma patients, concrete grammar is as follows:
Detect USP8 gene or its Partial Fragment sequence in ACTH type pituitary adenoma patients tumour to be measured,
If in described ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then ACTH type pituitary adenoma patients tumour to be measured is or candidate is ACTH type pituitary adenoma saltant type; If in described ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with described USP8 gene full wild-type sequence or corresponding part fragment sequence, then ACTH type pituitary adenoma patients tumour to be measured is or candidate is ACTH type pituitary adenoma wild-type;
USP8 gene wild-type sequence is sequence 1 in sequence table.
In above-mentioned detection ACTH to be measured type pituitary adenoma patients tumour USP8 gene or its Partial Fragment sequence method as follows:
1) USP8 gene or its Partial Fragment sequence in direct Sequencing ACTH type to be measured pituitary adenoma patients tumour;
2) to increase respectively USP8 gene different piece fragment in ACTH type pituitary adenoma patients tumour to be measured with each primer pair in primer set;
Primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
The diameter of tumor of ACTH type pituitary adenoma mutant subtypes is more than or equal to 1cm, and sudden change hypotype ACTH pituitary adenoma hypotype diameter of tumor is less than 1cm.
Whether whether embodiment 2, USP8 gene suddenly change at auxiliary detection pituitary adenoma patients to be measured is the application in ACTH type pituitary adenoma patients
Carry out USP8 detection in Gene Mutation through 120 kinds of ACTH type pituitary adenoma patients tumours of the non-ACTH type pituitary adenoma patients of pathology detection (comprising without hormone-type, lactotropin type, each 50 examples of tethelin type) tumour and embodiment 1 to 150 examples, concrete grammar is as follows:
Extract the genomic dna of each pituitary adenoma patients tumour, carry out pcr amplification respectively with the primer pair 1-3 of embodiment 1, order-checking pcr amplification product.
Result is as follows:
The nucleotide sequence of the pcr amplification product of 150 routine non-ACTH type pituitary adenoma patients tumours is consistent with the USP8 full length gene shown in sequence 1 or its Partial Fragment sequence, and the USP8 gene of 150 routine non-ACTH type pituitary adenoma patients tumours is wild-type;
In 120 kinds of ACTH type pituitary adenoma patients of embodiment 1,75 examples are ACTH type pituitary adenoma sudden change subgroups, account for 62.5% of sum.
Therefore, whether can be ACTH type by USP8 gene auxiliary detection pituitary adenoma patients tumour to be measured of whether undergoing mutation, concrete grammar be as follows:
To detect in pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then pituitary adenoma patients tumor candidate to be measured is ACTH type; If in described pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with USP8 gene full wild-type sequence or corresponding part fragment sequence, then pituitary adenoma patients tumor candidate to be measured is non-ACTH type; USP8 gene wild-type sequence is sequence 1 in sequence table.
In aforesaid method, in auxiliary detection pituitary adenoma patients to be measured tumour USP8 gene or its Partial Fragment sequence method as follows:
1) USP8 gene or its Partial Fragment sequence in direct Sequencing pituitary adenoma patients tumour to be measured;
2) to increase respectively USP8 gene different piece fragment in pituitary adenoma patients tumour to be measured with each primer pair in primer set;
Primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
Claims (8)
1., for detecting the test kit of ACTH type pituitary adenoma patients tumors subtypes to be measured, comprise the comparison card and sequenator of recording examination criteria;
Described examination criteria is as follows:
If in ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then ACTH type pituitary adenoma patients tumour to be measured is ACTH type pituitary adenoma saltant type or candidate is ACTH type pituitary adenoma saltant type; If in ACTH type pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with described USP8 gene full wild-type sequence or corresponding part fragment sequence, then ACTH type pituitary adenoma patients tumour to be measured is ACTH type pituitary adenoma wild-type or candidate is ACTH type pituitary adenoma wild-type;
Described USP8 gene wild-type sequence is sequence 1 in sequence table.
2. test kit according to claim 1, is characterized in that: described test kit also comprises the primer set for increase USP8 full length gene or its Partial Fragment;
Described primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
3., for a primer set for auxiliary detection ACTH type to be measured pituitary adenoma patients tumors subtypes, be made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
4. test kit described in claim 1 or 2 or the application of primer set according to claim 3 in the product preparing detection or auxiliary detection ACTH type to be measured pituitary adenoma patients tumors subtypes.
5. be whether the test kit of ACTH type for auxiliary detection pituitary adenoma patients tumour to be measured, comprise the comparison card and sequenator of recording examination criteria;
Described examination criteria is as follows:
If in pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence and USP8 gene full wild-type sequence or corresponding part fragment sequence inconsistent, then pituitary adenoma patients tumor candidate to be measured is ACTH type pituitary adenoma; If in pituitary adenoma patients tumour to be measured USP8 full length gene or its Partial Fragment sequence consistent with described USP8 gene full wild-type sequence or corresponding part fragment sequence, then pituitary adenoma patients tumor candidate to be measured is non-ACTH type pituitary adenoma;
Described USP8 gene wild-type sequence is sequence 1 in sequence table.
6. test kit according to claim 5, is characterized in that: described test kit also comprises the primer set for increase USP8 full length gene or its Partial Fragment;
Described primer set is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
7. whether auxiliary detection pituitary adenoma patients tumour to be measured is a primer set for ACTH type, is made up of primer pair 1, primer pair 2 and primer pair 3;
Described primer pair 1 is made up of the single strand dna shown in sequence 3 in the single strand dna shown in sequence in sequence table 2 and sequence table;
Described primer pair 2 is made up of the single strand dna shown in sequence 5 in the single strand dna shown in sequence in sequence table 4 and sequence table;
Described primer pair 3 is made up of the single strand dna shown in sequence 7 in the single strand dna shown in sequence in sequence table 6 and sequence table.
8. whether test kit described in claim 4 or 5 or primer set according to claim 7 are the application in the product of ACTH type in preparation auxiliary detection pituitary adenoma patients tumour to be measured.
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| CN105506116B (en) * | 2016-01-05 | 2019-01-08 | 上海交通大学 | BRAF gene detectable substance is preparing the application in ACTH type pituitary adenoma molecular diagnosis and parting product |
| CN106434918A (en) * | 2016-09-26 | 2017-02-22 | 复旦大学附属华山医院 | Application of specific gene mutation in molecular diagnosis and targeted therapy of pituitary adenoma detection |
| CN106381337A (en) * | 2016-10-27 | 2017-02-08 | 中国林业科学研究院林业新技术研究所 | Method for analyzing otis tarda food source plant through feces DNA (Deoxyribonucleic Acid) |
| CN106381337B (en) * | 2016-10-27 | 2019-11-19 | 中国林业科学研究院林业新技术研究所 | A method of great bustard food source plant is analyzed by faeces DNA |
| CN108624683A (en) * | 2017-03-24 | 2018-10-09 | 上海交通大学 | Application of the USP48 gene mutations in the diagnosis of ACTH type pituitary adenoma molecules |
| CN108624683B (en) * | 2017-03-24 | 2023-01-03 | 上海交通大学 | Application of USP48 gene mutation in ACTH-type pituitary adenoma molecular diagnosis |
| CN107937547A (en) * | 2018-01-04 | 2018-04-20 | 大连医科大学附属第二医院 | Circular rna DGKB is applied and expressed in pituitary adenoma biomarker |
| CN108220440A (en) * | 2018-01-04 | 2018-06-29 | 大连医科大学附属第二医院 | Application and expression of the NFKBIZ genes in pituitary adenoma biomarker |
| CN113621701A (en) * | 2020-05-06 | 2021-11-09 | 北京市神经外科研究所 | Application of estrogen receptor-associated receptor gamma (ESRRG) variant transcript in auxiliary diagnosis of pituitary adenoma |
| CN113005147A (en) * | 2021-03-11 | 2021-06-22 | 中山大学附属第一医院 | Construction method and application of mouse animal model with USP8 mutation |
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