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Lymphocyte gene rearrangement clonality assay kit and assay method thereof

A technology of gene rearrangement and lymphocytes, which is applied in the field of biological reagents, can solve the problems of high detection system cost, high false negative and false positive rates, and high cost of kits, so as to facilitate popularization and application, improve sensitivity and specificity, The effect of improving sensitivity and specificity

Inactive Publication Date: 2012-07-25
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The present inventor aims at the incomplete primer coverage, high false negative and false positive rates, high cost of existing kits and high cost of the detection system in the current domestic detection technology for gene rearrangement of lymphocytes. General hospital popularization and other issues, after extensive and in-depth research, and through repeated experiments to design a combined primer system, quickly and simply detect the clonality of lymphocyte gene rearrangement through ordinary PCR technology, and determine the proliferation of lymphoid tissue Benign and malignant lesions

Method used

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  • Lymphocyte gene rearrangement clonality assay kit and assay method thereof
  • Lymphocyte gene rearrangement clonality assay kit and assay method thereof
  • Lymphocyte gene rearrangement clonality assay kit and assay method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1: A detection kit for gene rearrangement clonality of lymphoid group cells

[0097] A practical and convenient detection kit for detecting gene rearrangement clonality of lymphoid group cells, said kit contains a plurality of independent containers or wells separated from each other, and each container or well is equipped with the following components respectively: Primer combination consisting of two primers:

[0098] Combination 1: forward primer indicated by SQE ID NO.1 + reverse primer indicated by SQE ID NO.2.

[0099] Combination 2: forward primer indicated by SQE ID NO.3 + reverse primer indicated by SQE ID NO.4.

[0100] Combination 3: forward primer indicated by SQE ID NO.5 + reverse primer indicated by SQE ID NO.6.

[0101] Combination 4: the forward primer shown in SQE ID NO.7 + the reverse primer shown in SQE ID NO.8.

[0102] Combination 5: forward primer indicated by SQE ID NO.9+reverse primer indicated by SQE ID NO.10.

[0103] Combination 6:...

Embodiment 2

[0114] Example 2: Determination of the clonality of lymphoid tissue gene rearrangement in the tissue to be tested

[0115] 1. Design of specific primer synthesis and detection method (such as Figure 1-9 shown)

[0116] According to the selected 14 gene rearrangements, combined with references, 14 sets of primers were designed using Oligo 6.2 primer design software to form 14 combinations, and the primer system was commissioned to a professional company (Shanghai Sunny Biotechnology Co., Ltd.) to synthesize.

[0117] In the primer system, the ratio of the two primers is 1:3-3:1, and the concentration is 10 pmol / ul.

[0118] (1) Combination 1:

[0119] SEQ ID NO: 1 5'-GGCCTCAGTGAAGGTCTCCTGCAAG-3'

[0120] SEQ ID NO: 2 3'-CCAGTGGCAGAGGAGTCCATTC-5'

[0121] Product size range 310-360bp

[0122] (2) Combination 2:

[0123]SEQ ID NO: 3 5'-CTGGGTGCGACAGGCCCCTGGACAA-3'

[0124] SEQ ID NO: 4 3'-CCAGTGGCAGAGGAGTCCATTC-5'

[0125] Product size range 250-290bp

[0126] (3) Combi...

Embodiment 3

[0188] Embodiment 3: Detection example one of the kit of the present invention

[0189] When B-cell lymphoma is clinically suspected: ① First, select combination 1+combination 2+combination 3+combination 6+combination 7 for PCR reaction, when a single PCR reaction product corresponding to any one of the selected combinations appears If there is no clonal band, it is diagnosed as B lymphocyte-derived lymphoma; ②When no monoclonal band appears, select combination 4+combination 5+combination 8 for PCR reaction. When monoclonal bands appear in the combined PCR reaction product, it is diagnosed as B-lymphocyte-derived lymphoma; ③ when no monoclonal bands appear, the possibility of B-cell lymphoma can be ruled out.

[0190] The specific cases are as follows:

[0191] ①Medical history: 53 / F, found a mass in the right cheek area for half a year, gradually enlarged

[0192] ②Pathological examination: a piece of parotid gland tissue on the right side, with a size of 4.5×3×2cm, and an ...

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Abstract

The invention belongs to the technical field of biological reagents, and relates to the assay of gene rearrangement clonality related to lymphoid neoplasm diagnosis. The development of a full-primer coverage, low-cost and easy-to-operate lymphoid tissue gene rearrangement assay kit capable of being popularized in ordinary hospitals is urgently needed clinically. The invention discloses a lymphocyte gene rearrangement clonality assay kit, which is all-liquid reagent based on combined primers, and the invention also provides a related assay method and a hierarchical optimization scheme. When the kit is used for assaying the clonality of lymphocyte gene rearrangement, the assay accuracy can reach more than 99 percent, moreover, the steps of the assay method are simple, the total cost is low, and the invention is highly worth popularizing clinically.

Description

technical field [0001] The invention belongs to the technical field of biological reagents, and in particular relates to a detection kit for gene rearrangement clonality of lymphocytes and a detection method thereof. Background technique [0002] Lymphoma is a common malignant tumor caused by the proliferation of lymphoid tissue. In recent years, with the popularization of industrialization and the aggravation of environmental pollution, especially the aggravation of indoor and outdoor decoration pollution, its incidence has increased year by year, seriously threatening human health. and quality of life. Although some progress has been made in the treatment of lymphoma in recent years, due to the difficulty in early diagnosis and the complexity of lymphoma types, the survival rate has not been fundamentally improved. Many patients are already in the critical stage of the disease when they are diagnosed In the late stage, the best chance of treatment is lost. [0003] Lymph...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 何妙侠倪灿荣郑建明赵静曹欢欢王建军白辰光马大烈
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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