Detection method for thyroid cancer related gene fusion mutation and kit
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A detection kit and gene fusion technology, applied in the field of genetic engineering, can solve the problems of long detection cycle, high cost, and low detection resolution
Pending Publication Date: 2020-06-30
深圳市新合生物医疗科技有限公司
View PDF3 Cites 3 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
[0007] Traditional detection methods for gene rearrangement include chromosome banding, fluorescence in situ hybridization and other techniques, but these techniques are time-consuming and laborious, and the detection reso
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more
Image
Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Embodiment 1
[0046] Embodiment 1 Detection kit for thyroid cancer-related gene fusion mutation based on the detection method for thyroid cancer-related gene fusion mutation provided by the present invention:
[0047] The detection kit includes the reagents required for RT-PCR detection, thyroid cancer-related fusion mutation gene specific primers, internal reference gene primers, negative RNA controls, and positive RNA controls.
[0048] The thyroid cancer-related gene fusion mutations include RET / PTC1 fusion, RET / PTC3 fusion and PAX8-PPARG rearrangement.
[0049] The reagents required for the above RT-PCR detection include SuperScript TM IV reverse transcriptase and 2×PlatinumSuperFi II PCR reaction mixture (product of Invitrogen company).
[0050] Thyroid cancer-related fusion mutant gene-specific primers and internal reference gene primers are the primers shown in SEQ ID NO. 1-12.
[0051] The above-mentioned negative RNA control substance is human total RNA that does not carry the a...
Embodiment 2
[0053] Embodiment 2 A detection method for thyroid cancer-related gene fusion mutation (using the detection kit described in Embodiment 1):
[0054] (1) Extract total RNA from the test sample;
[0055] The test sample was a paraffin-embedded sample (product number: 0710-0496) containing the three gene fusion mutations purchased from SeraCare Company in the United States. Using Quick-DNA / RNA from Zymo Research, USA TM FFPE Kits (Product No.: R1009) were used to extract total RNA from paraffin samples. The extraction method was carried out according to the instructions of the kit.
[0056] The extracted RNA was FFPE sample RNA containing RET / PTC1, RET / PTC3, PAX8 / PPARg.
[0057] At the same time, a negative control group was set up, and a negative control RNA sample was taken, which was purchased from Thermo Fisher (product number 361787), which did not contain any of RET / PTC1, RET / PTC3, and PAX8 / PPARg.
[0058] (2) The above-mentioned FFPE samples and negative control RNA s...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
PUM
Login to view more
Abstract
The invention provides a detection method for thyroid cancer related gene fusion mutation and a kit and relates to the technical field of gene engineering. The detection method comprises the followingsteps: performing detection, namely extracting total RNA (ribonucleic acid) or miRNA (micro ribonucleic acid) from a detection sample; performing reverse transcription on the total RNA or mRNA of thesample so as to obtain cDNA (complementary deoxyribonucleic acid), and performing fusion gene target specific PCR (polymerase chain reaction) amplification by using fusion gene target specific PCR amplification primers; performing electrophoresis detection on specific PCR amplification products so as to obtain types and sizes of fragments of the amplification products; and according to electrophoresis results, judging whether sample genes have thyroid cancer related gene fusion mutation or not. By adopting the method provided by the invention, three types of common gene fusion in thyroid cancer can be simply, conveniently and rapidly detected at an RNA level, and the method is high in sensitivity and applicable to clinical use.
Description
technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a detection method and kit for thyroid cancer-related gene fusion mutation. Background technique [0002] Thyroid cancer is the most common endocrine malignancy, and its incidence has continued and sharply increased in recent years. Timely and accurate identification of thyroid cancer is necessary. Clinical data statistics show that the detection rate of thyroid nodules by ultrasonography in the general population is about 35%. Ultimately, 80-95% of these nodules are identified as benign nodules, and only 5% to 15% of nodules. For malignant nodules, namely thyroid cancer. Therefore, it is necessary to further distinguish benign and malignant thyroid nodules. Ultrasound-guided fine-needle aspiration biopsy can ensure preoperative diagnosis of thyroid lesions. However, in 20–30% of nodule aspirates, cytology results are "inconclusive," and the risk of malignancy ran...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more
Application Information
Patent Timeline
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more