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Peripheral blood TCR marker of melanoma as well as detection kit and application of peripheral blood TCR marker

A technology of melanoma and markers, applied in the field of peripheral blood TCR markers and its detection kits, can solve the problems of difficult interpretation of test results, achieve low labor costs, simple sampling, and reduce testing costs

Pending Publication Date: 2020-09-22
CHENGDU EXAB BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pathological diagnosis must be combined with clinical evidence, and a comprehensive understanding of the patient's medical history and imaging examination information is required. For cases that are difficult to diagnose, more expert consultations should be invited, and the comprehensive evaluation of various indicators has caused serious problems for the interpretation of the final test results. difficulty

Method used

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  • Peripheral blood TCR marker of melanoma as well as detection kit and application of peripheral blood TCR marker
  • Peripheral blood TCR marker of melanoma as well as detection kit and application of peripheral blood TCR marker
  • Peripheral blood TCR marker of melanoma as well as detection kit and application of peripheral blood TCR marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1 Obtaining a melanoma TCR marker CDR3 sequence set through immunoassay analysis

[0053] 1. Sampling and immune map analysis (method reference patent ZL201910300069.9)

[0054] Collect 1301 control groups (including healthy people and patients with non-tumor diseases, 1300 people for model building, 1 healthy person for validation), 21 melanoma patients (20 people for model building, 1 person for validation) and the peripheral blood of a subject with unknown health status (10mL per person), and the amino acid sequence of the TCR epitope 3 (CDR3) of the subject and the control group was obtained by high-throughput sequencing to ensure the functionality of each sample The total number of CDR3 sequences of TCR should not be less than 30,000;

[0055] 2. The CDR3 sequence of the TCR of each sample is randomly sampled without replacement, so that the total number of CDR3 sequences of each sample is 30,000. For any specific CDR3 sequence X, the number of repeated o...

Embodiment 2

[0061] Example 2 Verification of the specificity of the melanoma TCR marker CDR3 sequence set

[0062] 1. Sampling and immune map analysis (method reference patent ZL201910300069.9)

[0063] The peripheral blood (10 mL per person) of 325 non-melanoma tumor patients and 5 subjects with unknown health status was collected, and the epitope 3 (CDR3) amino acid of the TCR of the subjects and the control group was obtained by high-throughput sequencing Sequences, to ensure that the total number of CDR3 sequences of functional TCRs in each sample is not less than 30,000; random non-replacement sampling is performed on the CDR3 sequences of TCRs in each sample, so that the total number of CDR3 sequences in each sample is 30,000.

[0064] 2. Randomly select 100 healthy people and 45 non-tumor disease patients from the control group in Example 1.

[0065] 3. According to the results of 100 healthy people, 45 non-tumor disease patients, and 20 melanoma patients from Example 1, and 325 n...

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Abstract

The invention discloses a peripheral blood TCR marker of melanoma as well as a detection kit and application of the peripheral blood TCR marker. The marker comprises at least one of proteins of whichthe sequences are shown as SEQ ID NO. 1-100. The sequence is based on a high-throughput sequencing method; only a small amount of peripheral blood needs to be collected; RNA (Ribonucleic Acid) is extracted, a sample is treated to establish an immune map library, high-throughput sequencing and TCR data analysis are carried out, a characteristic TCR sequence in peripheral blood of melanoma is determined, and then a test result of a sample to be tested is compared with the characteristic TCR sequence so as to determine whether the melanoma exists or not. According to the kit, a huge number of melanoma specific TCR sequences can be compared at the same time, and compared with single detection of one or more markers, the kit has higher specificity and accuracy, and the diagnosis efficiency is improved.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a peripheral blood TCR marker of melanoma, a detection kit and application thereof. Background technique [0002] Melanoma, also known as malignant melanoma, is a skin tumor related to orthopedics. Benign melanoma, also known as pigmented nevus, usually goes unnoticed, but melanoma can be malignant at the beginning, usually from junctional nevus. Although melanoma is a rare malignant tumor in my country, its mortality rate is high and its incidence is increasing year by year. Melanoma in my country is quite different from that of Caucasians in Europe and the United States in terms of pathogenesis, biological behavior, histological morphology, treatment methods, and prognosis. Among Asians and other people of color, about 50% of melanomas originate in the extremities, and the common primary sites are mostly found in extremities such as the soles, toes, fi...

Claims

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Application Information

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IPC IPC(8): G01N33/574A61K39/00A61P35/00
CPCG01N33/57407G01N33/57488A61K39/001111A61P35/00G01N2333/7051
Inventor 张志新杨鑫卓越文学平何彭铭
Owner CHENGDU EXAB BIOTECH CO LTD
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