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Sarcoma fusion gene detection kit and system

A detection kit and gene fusion technology, applied in the field of medical detection, can solve the problems of inconsistency in diagnosis, difficulty in diagnosis, affecting treatment plan formulation and prognosis judgment, etc.

Inactive Publication Date: 2019-05-28
GUANGZHOU KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The gold standard for the diagnosis of soft tissue sarcomas is histopathological diagnosis, however, similar morphology exists between a variety of benign and malignant tumors and between different sarcoma subtypes, which makes diagnosis difficult
It has been reported that nearly 25% of the cases were referred to the sarcoma diagnosis and treatment center with obvious diagnostic inconsistencies, and more than half of these cases affected the formulation of treatment plans and prognosis

Method used

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  • Sarcoma fusion gene detection kit and system
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  • Sarcoma fusion gene detection kit and system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] A sarcoma fusion gene detection kit, comprising the following primer sets:

[0056] Table 3. Fusion gene pairs and primer sets

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064] The above primer pair is one equivalent primer pair corresponding to each fusion site, such as COL1A1-PDGFB.C15P2.COSF1293 and COL1A1-PDGFB.C16P2.COSF595 and COL1A1-PDGFB.C16P2.COSF596 use the same primer pair, you need to follow SEQ 3 times of the primer pair of ID No.1-2 was added to the primer pair of SEQID No.9-10.

[0065] The kit of the present embodiment also includes the following internal reference gene primer sets:

[0066] Table 4. Internal reference gene primer set

[0067]

Embodiment 2

[0069] A sarcoma fusion gene detection system, comprising the following modules:

[0070] The detection module adopts the kit of Example 1 for high-throughput sequencing;

[0071] The analysis module obtains the above sequencing results, and obtains the fusion gene detection results through bioinformatics analysis;

[0072] The quality control module is verified by the predetermined quality control program. If the verification is passed, the test result will be output. If the verification fails, the sample will be invalid.

[0073] The workflow of the above detection system is as follows:

[0074] 1. Detection.

[0075] 1. RNA extraction

[0076] RNA was extracted from FFPE (formalin-fixed, paraffin-embedded) tissues.

[0077] 1.1 Preparation before experiment

[0078] a. Unless otherwise specified, all centrifugation is carried out at room temperature, and the centrifugation speed is 13200rpm;

[0079] b. FFPE tissue dosage: 4 pieces of 10μm thickness, surface area 2 (1...

Embodiment 3

[0201] Methodological validation of fusion gene detection in sarcoma.

[0202] 1. Sample source.

[0203] 1. 18 samples from the CAP Sarcoma Translocation PT project, for the official (CAP) confirmation of the status of the fusion gene of the sample.

[0204] 2. Clinical collection of Sarcoma FFPE samples: a total of 21 cases, 1 to 3 slides, all samples have been tested by FISH, and the test results are shown as follows:

[0205] Table 10. Information about 21 clinical SARC FFPE samples

[0206]

[0207]

[0208] The criteria for judging the above FISH test results are as follows:

[0209] 1) Analyze 200 cells, if the number of cells with positive signal is greater than 25%, it is judged to be positive for rearrangement;

[0210] 2) If the number of cells with positive signal is less than 10%, it is judged as rearrangement negative;

[0211] 3) Samples with a positive signal between 10% and 25% are re-tested; samples with a positive signal greater than 15% in the sec...

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Abstract

The invention relates to a sarcoma fusion gene detection kit and a system, and belongs to the technical field of medical detection. The kit carries out detection on 22 pairs of fusion genes (137 fusion transcripts) related to sarcoma, can be used for assisting in pathological diagnosis of soft tissue sarcoma and further can be used for carrying out prompting on targeted drug application.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a sarcoma fusion gene detection kit and system. Background technique [0002] Soft tissue sarcoma is a rare malignant tumor derived from stromal cells (mesemchymal cells), which may occur in any part of the body and has a wide age distribution. Soft tissue sarcoma accounts for about 1% of malignant tumors in adults, but accounts for 15% of malignant tumors in children. It is one of the most common solid tumors in children. [0003] The gold standard for the diagnosis of soft tissue sarcomas is histopathological diagnosis; however, similar morphology exists between various benign and malignant tumors and between different sarcoma subtypes, which makes diagnosis difficult. Studies have reported that nearly 25% of the cases were referred to the sarcoma diagnosis and treatment center with obvious diagnostic inconsistencies, and more than half of these cases affected the fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
Inventor 黄晓强刘菲菲欧小华胡昌明徐艳艳赵薇薇
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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