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A kit for detecting cis-trans mutations, a sample processing method and a determination method

A kit and cis-trans technology, applied in the field of detection materials, can solve the problems of high cost, time-consuming and labor-consuming, cumbersome operation, etc., and achieve the effect of convenient operation, high cost performance and less manual operation

Active Publication Date: 2021-09-03
SHENZHEN YOU SHENGKANG BIOSCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Sanger sequencing technology has high detection specificity and throughput, but the method itself is not sensitive and cannot detect large fragment deletion rearrangements, and pre-PCR and electrophoresis are required for sequencing, which is time-consuming and labor-intensive, and is not suitable for large-scale application
The next-generation sequencing technology has greatly improved the sequencing depth, so the sensitivity has been improved compared with the first-generation sequencing, but the detection cost, sensitivity and convenience of operation are not as good as ARMS-PCR technology
The main disadvantages are high cost, cumbersome operation, and low throughput, and only one mutation site can be detected in one reaction

Method used

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  • A kit for detecting cis-trans mutations, a sample processing method and a determination method
  • A kit for detecting cis-trans mutations, a sample processing method and a determination method
  • A kit for detecting cis-trans mutations, a sample processing method and a determination method

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Experimental program
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Effect test

Embodiment 1

[0041] 1. Primer sequence design, see Table 1 for details:

[0042]

[0043]

[0044] 2. Sample preparation

[0045] Use the cfDNAkit to extract the plasma DNA of the patients with the kit.

[0046] 3. Instruments

[0047] ABI7500, high-speed centrifuge, low-speed centrifuge, constant temperature dry bath, vortex oscillator, refrigerator.

[0048] 4. The composition of the kit

[0049] The kit consists of EGFR 4-way PCR reaction strips and EGFR positive control.

[0050] The basic composition of each well of the reaction strip is: 1×PCR Buffer, 2~8pmol dNTPs, 1.5~4.0mMMgCl 2 ,, 0.5~2U HS Taq, 0.1~1U UNGase. According to Table 1), each primer probe was sequentially added to the 4-tube tube, and the final concentration was 100-400 nM.

[0051] 5. Testing process

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Abstract

The invention provides a kit for detecting cis-trans mutations, a sample processing method and a determination method, wherein the kit includes internal control primers and mutation detection primers. The sample processing method includes four steps of obtaining the sample to be tested, sample preparation, detection, and obtaining the processed sample. Judgment methods, including positive result judgment, T790M‑C797S cis-trans judgment and quality control. Beneficial effects: 1. High sensitivity; 2. Wide range of applicable samples; 3. Low cost; 4. Convenient operation.

Description

technical field [0001] The invention relates to a detection material, in particular to a kit for detecting cis-trans mutations, a sample processing method and a determination method. The invention can be used for detecting cis-trans mutations of lung cancer EGFR gene T790M, C797S and T790M-C797S. The invention is only used for auxiliary detection of drug guidance for lung cancer patients, not a disease diagnosis reagent. Background technique [0002] Studies have shown that cancer is the result of the interaction between environmental factors and cell genetic material, the result of multi-factor, multi-stage and multi-gene effects, and the result of the accumulation of gene mutations, so cancer is a genetic disease. Among the many human genes, proto-oncogenes and tumor suppressor genes are closely related to the occurrence and development of cancer. Mutations of proto-oncogenes and or tumor suppressor genes can cause cell canceration. Before cancer forms a solid mass, the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/156C12Q2600/166
Inventor 喻德华罗玄翟建新李鑫剩
Owner SHENZHEN YOU SHENGKANG BIOSCI CO LTD