CN109680068A - A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker - Google Patents
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker Download PDFInfo
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- CN109680068A CN109680068A CN201910157198.7A CN201910157198A CN109680068A CN 109680068 A CN109680068 A CN 109680068A CN 201910157198 A CN201910157198 A CN 201910157198A CN 109680068 A CN109680068 A CN 109680068A
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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
- 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
- C12Q2600/158—Expression markers
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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
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Abstract
The invention discloses a kind of methods using blood plasma miRNA marker diagnosis parathyroid carcinoma, are to solve the problem of parathyroid carcinoma clinically lacks effective preoperative diagnosis method.Specific step is as follows: step 1 by the present invention, extracts the blood of patient and is placed in anticoagulant tube, joins outside addition standard into anticoagulant tube, and centrifugation is to obtain blood plasma；Step 2 extracts the miRNA to dissociate in blood plasma；Step 3 is measured the content of the miRNA to dissociate in blood plasma, is standardized using outer parameter evidence and internal reference data, obtains testing result；Step 4 establishes parathyroid carcinoma predictor formula；Step 5 will test result and substitute into formula.The level that the present invention passes through one group of miRNA in detection blood plasma, parathyroid carcinoma predictor formula is established, a possibility that patient suffers from malignant tumour is calculated, to take corresponding remedy measures, it is horizontal that parathyroid carcinoma preoperative diagnosis can be significantly improved, provide objective basis to reasonably select modus operandi.
Description
Technical field
It is specifically a kind of to be diagnosed by first shape using blood plasma miRNA marker the present invention relates to parathyroid carcinoma diagnostic field
The method of gland cancer.
Background technique
Parathyroid carcinoma is a kind of poor pernicious endocrine tumors of prognosis, and major clinical feature is a large amount of secretion first shapes
Other glandular hormone causes patient uncontrollable hypercalcinemia occur, finally results in patient and cardiorespiratory failure occurs and lethal.First
The optimal treatment means of gland cancer are to take radical surgery, complete resection parathyroid gland lesion, week when performing the operation first time by shape
The thyroid gland enclosed and other soft tissues avoid tumour rupture residual or send out.But clinically lack effectively preoperative examine at present
Disconnected method, relies primarily on the clinical manifestation of pre-operative patients, is speculated according to the Tumors display in operation.As patient in the preoperative
Clinical symptoms weight, and serum PTH and blood calcium obviously increase, then suspect parathyroid carcinoma.And perform the operation in detect discovery tumour with
Surrounding tissue adhesion is serious and the performance of parathyroid carcinoma.But these show and do not have specificity, by some benign first shapes
Gland adenoma, especially gross tumor volume be larger or the longer case of the course of disease, is likely to occur above-mentioned performance.Frozen section pathology in art
Checking has good antidiastole effect for most of tumour, but lacks objective finger for the diagnosis of parathyroid carcinoma
Mark, accuracy is poor, and repeatability is poor, often occurs failing to pinpoint a disease in diagnosis or mistaken diagnosis.Even postoperative pathological diagnoses, due in institutional framework
Lack clarification of objective, most of patient, which still will appear, to fail to pinpoint a disease in diagnosis.For the parathyroid carcinoma patient failed to pinpoint a disease in diagnosis, after tumor recurrence
On the one hand row second operation again dramatically increases the risk of tumour metastasis and recurrence, still further aspect obviously increases operating difficulty, brings
The operation related complication such as recurrent nerve injury.It is such as parathyroid carcinoma by parathyroid gland benign tumour mistaken diagnosis, then causes to perform the operation
Excision extension is excessive, there are problems that over-treatment, brings thyroid function is low the risks such as to subtract to patient, while bringing to patient
Huge mental burden.
If can be diagnosed before surgery, the tumor operation of radical-ability is taken in time, then may improve the long-term of patient
Life span avoids unnecessary postoperative complication, while saving medical resource.
Summary of the invention
The purpose of the present invention is to provide a kind of methods using blood plasma miRNA marker diagnosis parathyroid carcinoma, with solution
Certainly the problems mentioned above in the background art.
To achieve the above object, the invention provides the following technical scheme:
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1, extract be determined as primary hyperparathyroidism patient blood and by the blood be placed in containing
In the anticoagulant tube of EDTA solution, then join outside addition standard into anticoagulant tube, be then centrifuged in centrifuge, obtains blood plasma；
Step 2, the miRNA to dissociate in blood plasma is extracted using miRNA extracts kit, and miRNA refers to tiny RNA；
Step 3 measures the content of the miRNA to dissociate in blood plasma, using outer parameter evidence and internal reference data to plasma results into
Row standardization, obtains testing result；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
As a further solution of the present invention: containing using the miRNA to dissociate in round pcr measurement blood plasma in step 3
Amount.
As a further solution of the present invention: the miRNA to dissociate in blood plasma includes hsa-miR-6870-3p, hsa-miR-
762、hsa-miR-4516、hsa-miR-3960、hsa-miR-6090、hsa-miR-6869-5p、hsa-miR-6088、hsa-
miR-638、hsa-miR-8069、hsa-miR-5703、hsa-miR-6089、hsa-miR-4281、hsa-miR-1225-5p、
hsa-miR-7107-5p、hsa-miR-7847-3p、hsa-miR-6510-5p、hsa-miR-7110-5p、hsa-miR-21-
5p, hsa-miR-1304-3p, hsa-miR-6889-3p, hsa-miR-6795-3p and hsa-miR-3162-3p.
As a further solution of the present invention: round pcr includes any one in qPCR technology and dPCR technology, qPCR
Technology is real time RT round pcr, and dPCR technology is digital pcr technology.
Compared with prior art, the beneficial effects of the present invention are:
The present invention establishes parathyroid carcinoma predictor formula, utilizes formula meter by the level of one group of miRNA in detection blood plasma
A possibility that patient suffers from malignant tumour is calculated, to take corresponding remedy measures；
It is horizontal that the present invention can significantly improve parathyroid carcinoma preoperative diagnosis, for reasonably select modus operandi provide it is objective according to
According to；
The present invention can be combined with molecular bar code technology and kit be made, and develop product, benefit many patients, have product
The social benefit of pole.
Specific embodiment
The technical solution of the patent is explained in further detail With reference to embodiment.
Embodiment 1
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1 extracts the blood 6mL for being determined as primary hyperparathyroidism patient and is placed in the blood
In anticoagulant tube containing EDTA solution, then join outside addition standard into anticoagulant tube, be then centrifuged in centrifuge, obtains blood plasma
3mL；
Step 2, the miRNA to dissociate in blood plasma is extracted using miRNA extracts kit, and the miRNA to dissociate in blood plasma includes
hsa-miR-6870-3p、hsa-miR-762、hsa-miR-4516、hsa-miR-3960、hsa-miR-6090、hsa-miR-
6869-5p、hsa-miR-6088、hsa-miR-638、hsa-miR-8069、hsa-miR-5703、hsa-miR-6089、hsa-
miR-4281、hsa-miR-1225-5p、hsa-miR-7107-5p、hsa-miR-7847-3p、hsa-miR-6510-5p、hsa-
MiR-7110-5p, hsa-miR-21-5p, hsa-miR-1304-3p, hsa-miR-6889-3p, hsa-miR-6795-3p and
hsa-miR-3162-3p；
Step 3 utilizes outer parameter evidence and internal reference data using the content of the miRNA to dissociate in round pcr measurement blood plasma
Plasma results are standardized, testing result is obtained；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
Embodiment 2
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1 extracts the blood 4mL for being determined as primary hyperparathyroidism patient and is placed in the blood
In anticoagulant tube containing EDTA solution, then join outside addition standard into anticoagulant tube, be centrifuged in centrifuge in 30 minutes, obtains blood
Starch 2mL；
Step 2 extracts the miRNA to dissociate in blood plasma using miRNA extracts kit；
Step 3 utilizes outer parameter evidence and internal reference data using the content of the miRNA to dissociate in qPCR technology measurement blood plasma
Plasma results are standardized, testing result is obtained；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
Embodiment 3
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1 extracts the blood 5mL for being determined as primary hyperparathyroidism patient and is placed in the blood
In anticoagulant tube containing EDTA solution, then join outside addition standard into anticoagulant tube, in the centrifugation that revolving speed is 2250rpm in 2 hours
It is centrifuged in machine, obtains blood plasma 2.5mL；
Step 2, the miRNA to dissociate in blood plasma is extracted using miRNA extracts kit, and the miRNA to dissociate in blood plasma includes
hsa-miR-6870-3p、hsa-miR-762、hsa-miR-4516、hsa-miR-3960、hsa-miR-6090、hsa-miR-
6869-5p、hsa-miR-6088、hsa-miR-638、hsa-miR-8069、hsa-miR-5703、hsa-miR-6089、hsa-
miR-4281、hsa-miR-1225-5p、hsa-miR-7107-5p、hsa-miR-7847-3p、hsa-miR-6510-5p、hsa-
MiR-7110-5p, hsa-miR-21-5p, hsa-miR-1304-3p, hsa-miR-6889-3p, hsa-miR-6795-3p and
hsa-miR-3162-3p；
Step 3 utilizes outer parameter evidence and internal reference data using the content of the miRNA to dissociate in dPCR technology measurement blood plasma
Plasma results are standardized, testing result is obtained；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
Embodiment 4
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1, extract be determined as primary hyperparathyroidism patient blood and by the blood be placed in containing
In the anticoagulant tube of EDTA solution, then join outside addition standard into anticoagulant tube, be then centrifuged in centrifuge, obtains blood plasma, -80 DEG C
Cryo-conservation 7 days；
Step 2, the miRNA to dissociate in blood plasma is extracted using miRNA extracts kit, and the miRNA to dissociate in blood plasma includes
hsa-miR-6870-3p、hsa-miR-762、hsa-miR-4516、hsa-miR-3960、hsa-miR-6090、hsa-miR-
6869-5p、hsa-miR-6088、hsa-miR-638、hsa-miR-8069、hsa-miR-5703、hsa-miR-6089、hsa-
miR-4281、hsa-miR-1225-5p、hsa-miR-7107-5p、hsa-miR-7847-3p、hsa-miR-6510-5p、hsa-
MiR-7110-5p, hsa-miR-21-5p, hsa-miR-1304-3p, hsa-miR-6889-3p, hsa-miR-6795-3p and
hsa-miR-3162-3p；
Step 3 utilizes outer parameter evidence and internal reference data using the content of the miRNA to dissociate in round pcr measurement blood plasma
Plasma results are standardized, testing result is obtained；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
Embodiment 5
A method of parathyroid carcinoma is diagnosed using blood plasma miRNA marker, the specific steps are as follows:
Step 1 extracts the blood 5mL for being determined as primary hyperparathyroidism patient and is placed in the blood
In anticoagulant tube containing EDTA solution, then join outside addition standard into anticoagulant tube, be then centrifuged in centrifuge, obtains blood plasma
2.5mL；
Step 2, the miRNA to dissociate in blood plasma is extracted using miRNA extracts kit, and the miRNA to dissociate in blood plasma includes
hsa-miR-6870-3p、hsa-miR-762、hsa-miR-4516、hsa-miR-3960、hsa-miR-6090、hsa-miR-
6869-5p、hsa-miR-6088、hsa-miR-638、hsa-miR-8069、hsa-miR-5703、hsa-miR-6089、hsa-
miR-4281、hsa-miR-1225-5p、hsa-miR-7107-5p、hsa-miR-7847-3p、hsa-miR-6510-5p、hsa-
MiR-7110-5p, hsa-miR-21-5p, hsa-miR-1304-3p, hsa-miR-6889-3p, hsa-miR-6795-3p and
hsa-miR-3162-3p；
Step 3 measures the content of the miRNA to dissociate in blood plasma, using outer parameter evidence and internal reference data to plasma results into
Row standardization, obtains testing result；
Step 4 establishes parathyroid carcinoma predictor formula, can be returned with logistic and the foundation of the methods of neural Meta algorithm
Formula.
Step 5 will test result and substitute into formula, can assess probability, postoperative recurrence that the patient suffers from parathyroid carcinoma and turn
The risk of shifting and the selection for assisting modus operandi.
Step 6 gradually accumulates data in use, is constantly opposed prediction with pathological diagnosis result and Follow-up Data
Formula is modified.
The working principle of the invention is: the present invention collects the plasma specimen of patient in the preoperative, passes through one group in detection blood plasma
The level of miRNA establishes parathyroid carcinoma predictor formula, a possibility that patient suffers from malignant tumour is calculated using formula, to adopt
Take corresponding remedy measures, the present invention establishes objectively repeatable molecular diagnosis method, provides by a kind of preoperative diagnosis first shape
The Diagnostic Strategy of gland cancer, to significantly improve the diagnostic accuracy of preoperative parathyroid carcinoma, to select optimal hand for patient
Art mode improves the quality of life and life span of patient.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
In addition, it should be understood that although this specification is described in terms of embodiments, but not each embodiment is only wrapped
Containing an independent technical solution, this description of the specification is merely for the sake of clarity, and those skilled in the art should
It considers the specification as a whole, the technical solutions in the various embodiments may also be suitably combined, forms those skilled in the art
The other embodiments being understood that.
Claims (5)
1. a kind of method using blood plasma miRNA marker diagnosis parathyroid carcinoma, which is characterized in that specific step is as follows:
Step 1 extracts the blood for being determined as primary hyperparathyroidism patient and the blood is placed in containing EDTA
In the anticoagulant tube of solution, then join outside addition standard into anticoagulant tube, be then centrifuged in centrifuge, obtains blood plasma；
Step 2 extracts the miRNA to dissociate in blood plasma using miRNA extracts kit；
Step 3 is measured the content of the miRNA to dissociate in blood plasma, is marked using outer parameter evidence and internal reference data to plasma results
Standardization obtains testing result；
Step 4 establishes parathyroid carcinoma predictor formula；
Step 5 will test result and substitute into parathyroid carcinoma predictor formula.
2. the method according to claim 1 using blood plasma miRNA marker diagnosis parathyroid carcinoma, which is characterized in that
Using the content of the miRNA to dissociate in round pcr measurement blood plasma in the step 3.
3. the method according to claim 1 using blood plasma miRNA marker diagnosis parathyroid carcinoma, which is characterized in that
The miRNA to dissociate in the blood plasma includes hsa-miR-6870-3p, hsa-miR-762, hsa-miR-4516, hsa-miR-
3960、hsa-miR-6090、hsa-miR-6869-5p、hsa-miR-6088、hsa-miR-638、hsa-miR-8069、hsa-
miR-5703、hsa-miR-6089、hsa-miR-4281、hsa-miR-1225-5p、hsa-miR-7107-5p、hsa-miR-
7847-3p、hsa-miR-6510-5p、hsa-miR-7110-5p、hsa-miR-21-5p、hsa-miR-1304-3p、hsa-
MiR-6889-3p, hsa-miR-6795-3p and hsa-miR-3162-3p.
4. the method according to claim 2 using blood plasma miRNA marker diagnosis parathyroid carcinoma, which is characterized in that
The round pcr includes any one in real time RT round pcr and digital round pcr.
5. the method according to claim 1 using blood plasma miRNA marker diagnosis parathyroid carcinoma, which is characterized in that
The parathyroid carcinoma predictor formula uses the logistic Return Law or neural Meta algorithm.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103237901A (en) * | 2010-03-01 | 2013-08-07 | 卡里斯生命科学卢森堡控股有限责任公司 | Biomarkers for theranostics |
| US20130252832A1 (en) * | 2012-03-22 | 2013-09-26 | Yale University | KRAS Variant and Tumor Biology |
-
2019
- 2019-03-01 CN CN201910157198.7A patent/CN109680068A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103237901A (en) * | 2010-03-01 | 2013-08-07 | 卡里斯生命科学卢森堡控股有限责任公司 | Biomarkers for theranostics |
| US20130252832A1 (en) * | 2012-03-22 | 2013-09-26 | Yale University | KRAS Variant and Tumor Biology |
Non-Patent Citations (4)
| Title |
|---|
| YA HU等: "Verification of candidate microRNA markers for parathyroid carcinoma", 《ENDOCRINE GENETICS/EPIGENETICS》 * |
| 孔晶: "1、原发性甲状旁腺功能亢进症的外周血白细胞microRNA表达谱2、多发性内分泌腺瘤病1型相关的甲状旁腺肿瘤p27~（Kip1）及β-连环蛋白的表达", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 张向阳: "《医学分子生物学》", 28 February 2018, 江苏凤凰科学技术出版社 * |
| 谢宗燕等: "microRNA在甲状旁腺癌中的研究现状及其发展前景", 《中华内分泌代谢杂志》 * |
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Application publication date: 20190426 |