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Kit for evaluating effect of 131I on treating differentiated thyroid cancer

A thyroid cancer and kit technology, which is applied to the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of decreased iodine absorption capacity, no clear determination, and poor radiotherapy, etc. Achieve the effect of less non-specific amplification, overcoming mutual interference, and high uniformity

Inactive Publication Date: 2021-04-13
润安医学科技(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the "Guidelines for the Diagnosis and Treatment of Persistent / Recurrent and Metastatic Thyroid Cancer" (Chinese Society of Clinical Oncology (CSCO), 2018. V1) pointed out that the BRAF gene V600E mutation and TERT gene promoter mutation in the primary tumor of papillary thyroid carcinoma (PTC) It is related to the decline of iodine uptake ability of distant metastases, resulting in a decline of iodine absorption ability, and the possibility of benefit from iodine treatment is small, but so far these expert consensuses or diagnosis and treatment guidelines have not clarified which promoter regions are on the TERT gene Affects iodine uptake capacity
Through a large number of clinical studies, we found that patients with papillary thyroid carcinoma carrying TERT gene promoter C228T and C250T site mutations have reduced iodine absorption ability, and postoperative 131I radiation therapy is not good
[0006]In addition, although some studies have found that in follicular carcinoma of the thyroid with papillary nucleus features, patients with NRAS gene mutations received 131 The benefit of sup>I treatment is more likely, but whether other subtypes of RAS gene are related to the therapeutic effect of 131I is not yet clear.
After research, we found that the mutations in the G12 / G13 / Q61 region of the HRAS gene are closely related to the therapeutic effect of 131I, but the mutations in the KRAS gene are closely related to the therapeutic effect of 131I effect irrelevant

Method used

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  • Kit for evaluating effect of 131I on treating differentiated thyroid cancer
  • Kit for evaluating effect of 131I on treating differentiated thyroid cancer
  • Kit for evaluating effect of 131I on treating differentiated thyroid cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: A method for evaluating 131 The combination of primers for the effect of I in treating differentiated thyroid cancer

[0045] This embodiment involves with 131 The gene mutation sites related to the effect of I treatment of differentiated thyroid cancer were selected from the COSMIC (Catalogue of Somatic Mutations in Cancer) database, and primers were designed according to the relevant gene sequences. A total of 16 pairs of specific primers were designed, as shown in Table 2. Each pair of specific primers amplifies the target region with a size of 180-200bp, and the size of the amplified product is 220-280bp. It has the advantages of wide coverage, many detection sites, balanced GC content, stable product structure, and less dimer structure. It exhibits good specificity, stability and uniformity in multiple PCR amplification, and can ensure the amplification efficiency while ensuring the specificity of PCR amplification.

[0046] Primers designed in this ex...

Embodiment 2

[0048] Example 2: A method for evaluating 131 Kit for the effect of I treatment of differentiated thyroid cancer

[0049] The one described in this example is used to evaluate 131 I The kit for treating differentiated thyroid cancer effect, its design principle is as follows figure 1 As shown, it mainly includes:

[0050] (1) PCR-specific primers: used to amplify multiple target regions on the target gene of the sample to be tested. The amplification range covers at least the hotspot mutation region of the target gene. The sequences are as shown in Table 2. SEQ ID NO.1 to SEQ ID NO .12. Preferably, multiple amplification primer pairs are mixed together to form a primer pool;

[0051] (2) Universal primers: used to re-amplify the amplification products of the target region amplified by specific primers during the library construction process, to mark the sequencing libraries of different samples to be tested, and then to distinguish different samples. The sequence is shown...

Embodiment 3

[0055] Example 3: A method for evaluating 131 Detection process of the kit for the effect of I treatment of differentiated thyroid cancer

[0056] The detection process of this embodiment is as follows figure 2 shown, including the following steps:

[0057] 1) Sample DNA extraction

[0058] Use the QIAamp DNA Mini Kit (51304) kit to extract fresh tissue DNA. The specific steps are operated according to the kit instructions. After the DNA is extracted, use a spectrophotometer for quality control analysis. The value of A260 / A280 is between 1.9-2.1, using Qubit Concentration determination was performed with a fluorescence quantitator; the extracted DNA was diluted to 10-50 ng / μL using nuclease-free water as a template.

[0059] 2) The first round of ligation PCR

[0060] components volume High Fidelity PCR Enzyme Mix 12.5μL DNA-specific primer mix pool 4μL DNA template 2μL nuclease free water Make up to 25 μL

[0061] DNA multiple...

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Abstract

The invention discloses a primer and kit for evaluating the effect of 131I on treating differentiated thyroid cancer. The kit can be used for simultaneously detecting the variation of nine gene loci of a BRAF gene, a TERT gene, an HRAS gene and an NRAS gene, and the variation of the gene loci is closely related to the treatment effect of 131I on differentiated thyroid cancer. Therefore, when the kit is used for detecting a sample of a patient, a doctor can be assisted in scientifically evaluating the clinical effect of treating a differentiated thyroid cancer patient by using 131I, a basis is provided for making a precise treatment scheme of the patient, and the kit undoubtedly has very important clinical use value.

Description

technical field [0001] The invention relates to the field of molecular biology technology, in particular to a method for evaluating 131 I kit for treating differentiated thyroid cancer efficacy. Background technique [0002] Papillary thyroid carcinoma (PTC) and follicular thyroid carcinoma (FTC) account for more than 90% of thyroid cancers, because they retain the function of thyroid follicular epithelial cells to a certain extent, such as sodium The expression of iodide transporter (sodium iodide symporter, NIS), the ability of iodine uptake, the ability of secreting thyroglobulin (thyroglobulin, Tg), the growth mode dependent on thyroid-stimulating hormone (thyroid-stimulating hormone, TSH), etc. It is called differentiated thyroid cancer (differentiated thyroid cancer, DTC). The increased incidence of DTC, especially PTC, is the main factor leading to the increase in the incidence of thyroid cancer in recent years. 131 I treatment of differentiated thyroid cancer (DTC)...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6869C12N15/11
CPCC12Q1/6886C12Q1/6869C12Q2600/156C12Q2600/106C12Q2535/122
Inventor 宣涛张宇清裴婷婷
Owner 润安医学科技(苏州)有限公司
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