Peripheral blood TCR marker of acute B lymphocytic leukemia as well as detection kit and application of peripheral blood TCR marker

A B lymphocyte and marker technology, applied in the field of genetic engineering, can solve the problems of false negative results, insufficient sensitivity, and easy results, and achieve low labor cost, high specificity and accuracy, and simple sampling. Effect

Pending Publication Date: 2021-06-25
CHENGDU EXAB BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of detection has a fast measurement speed and can measure multiple parameters, but its sensitivity is not enough, and antigenic drift may occur in patients with minimal residual disease in ALL patients, resulting in false negative results of flow cytometry
[0007] PCR includes real-time fluorescent quantitative RCR (qPCR) and reverse transcription PCR (RT-PCR), both of which have high sensitivity for detecting minimal residual disease, but the results are prone to false negatives

Method used

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  • Peripheral blood TCR marker of acute B lymphocytic leukemia as well as detection kit and application of peripheral blood TCR marker
  • Peripheral blood TCR marker of acute B lymphocytic leukemia as well as detection kit and application of peripheral blood TCR marker
  • Peripheral blood TCR marker of acute B lymphocytic leukemia as well as detection kit and application of peripheral blood TCR marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Obtaining Acute B Lymphocytic Leukemia TCR Marker CDR3 Sequence Set by Immunogram Analysis

[0036] 1. Sampling and immune map analysis

[0037] Collect 1301 control groups (including healthy people and patients with non-tumor diseases, 1300 people for model building, 1 healthy person for verification), 56 patients with acute B-lymphoblastic leukemia (55 people for model building, 1 person for for verification) and the peripheral blood of a subject with unknown health status (10mL per person), the amino acid sequence of the TCR epitope 3 (CDR3) of the subject and the control group was obtained by high-throughput sequencing, ensuring that each sample The total number of CDR3 sequences of the functional TCR is not less than 25,000;

[0038]2. Perform random non-replacement sampling on the sequences of samples whose total number of CDR3 sequences of each functional TCR is higher than 30,000, so that the total number of CDR3 sequences of this sample is 30,000. S...

Embodiment 2

[0044] Example 2 Verification of the specificity of the TCR marker CDR3 sequence set in acute B-lymphoblastic leukemia

[0045] 1. Sampling and immune map analysis

[0046] Peripheral blood (10 mL per person) was collected from 626 tumor patients with non-acute B-lymphoblastic leukemia and 7 subjects with unknown health status, and the epitope 3 ( CDR3) amino acid sequence, to ensure that the total number of CDR3 sequences of each functional TCR in each sample is not less than 25,000; random non-replacement sampling is performed on the sequences of samples whose total number of CDR3 sequences in each functional TCR is higher than 30,000. The total number of CDR3 sequences of the samples is 30000. So far, the total number of functional TCR CDR3 sequences contained in all samples is 25,000-30,000.

[0047] 2. Randomly select 97 healthy people and 41 non-tumor disease patients from the control group in Example 1.

[0048] 3. According to the 97 healthy people, 41 non-neoplasti...

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Abstract

The invention discloses a peripheral blood TCR marker of acute B lymphocytic leukemia as well as a detection kit and application of the peripheral blood TCR marker. The peripheral blood TCR marker comprises at least one of proteins of which the sequences are as shown in SEQ ID NO.1-100. On the basis of a high-throughput sequencing method, only a small amount of peripheral blood needs to be adopted, RNA is extracted, an immune map library is established by treating a sample, then through high-throughput sequencing and TCR data analysis, firstly, a characteristic TCR sequence in the peripheral blood of the acute B lymphocytic leukemia is determined, and then a test result of a to-be-tested sample is compared with the characteristic TCR sequence, therefore, whether a patient suffers from acute B lymphocytic leukemia is determined. According to the peripheral blood TCR marker, a huge number of acute B lymphocytic leukemia specific TCR sequences can be compared at the same time, and the peripheral blood TCR marker has higher specificity, higher accuracy and improved diagnosis efficiency compared with single detection of one or more markers.

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 acute B lymphocytic leukemia, a detection kit and application thereof. Background technique [0002] Acute Lymphoblastic Leukemia (ALL) is a malignant tumor disease originating from the abnormal proliferation of B-lineage or T-lineage lymphoid progenitor cells in the bone marrow. Abnormally proliferating blast cells can gather in the bone marrow and inhibit normal hematopoietic function, and can also invade tissues outside the bone marrow, such as meninges, lymph nodes, gonads, and liver. my country has conducted a survey on the incidence of leukemia, and the incidence of ALL is about 0.67 / 100,000. The incidence rate in oil fields and polluted areas is significantly higher than the national incidence rate. ALL childhood (0 to 9 years old) is the peak incidence, which can account for more than 70% of childhood leukemia. A...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/68A61K38/17A61P35/02C07K14/725
CPCG01N33/57426G01N33/57488G01N33/6893A61K38/1774A61P35/02C07K14/7051G01N2333/7051
Inventor 张志新卓越杨鑫
Owner CHENGDU EXAB BIOTECH CO LTD
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