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Detection probes and detection liquid phase chip for BIM gene deletion mutation

A gene deletion and mutation detection technology, applied in the field of molecular biology, can solve the problems of inappropriate clinical diagnostic chip, high detection price, low degree of automation, etc., achieve accurate and reliable detection results, overcome low sensitivity, and improve sensitivity Effect

Active Publication Date: 2014-02-12
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the Illumina fiber optic bead chip technology is a high-throughput detection system with high sensitivity and accuracy, it has a low degree of automation and many manual operations, which is difficult to meet the needs of practical applications. The high-throughput Affymetrix SNP6.0 chip technology is relatively mature. However, this chip technology is not suitable for low-to-medium density clinical diagnostic chips, and it is difficult to expand the detection of SNPs or tagged SNPs related to many biological traits in the same reaction system.
In addition, the Affymetrix SNP6.0 chip is mainly strong on the expression spectrum chip, with many species, relatively weak on the SNP chip, and the detection price is expensive, which cannot meet the actual needs

Method used

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  • Detection probes and detection liquid phase chip for BIM gene deletion mutation
  • Detection probes and detection liquid phase chip for BIM gene deletion mutation
  • Detection probes and detection liquid phase chip for BIM gene deletion mutation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1BI

[0023] Embodiment 1 BIM gene deletion mutation detection liquid chip mainly includes:

[0024] 1. Detection probe

[0025] For the wild-type and mutant types of the BIM gene deletion mutation detected by the target of the present invention, design detection probes, as shown in the following table:

[0026] Table 1 Detection probe sequences for BIM gene deletion mutations

[0027]

[0028]

[0029] The four selected microspheres were purchased from Luminex, USA, and the detection probes were coated on the microspheres (one probe corresponds to one microsphere). A 20-50 T spacer sequence is connected between the detection probe and the microsphere, that is, a 20-50 T spacer sequence is added before each detection sequence. The detection probe sequence is provided by Shanghai Sangon Bioengineering Co., Ltd. Technical Services Ltd Synthesis. The synthesized detection probe was treated with sterilized ddH 2 O was made into a stock solution of 100nmol / ml.

[0030] The spa...

Embodiment 3 Embodiment 1

[0077] BIM gene mutation detection detection probe in embodiment 3 embodiment 1

[0078] 1. Design of liquid phase chip preparation

[0079] 2 wild-type detection probes (SEQ ID NO.1 and SEQ ID NO.2) and mutant detection probes (SEQ ID NO.3 and SEQ ID NO. .4) to form different detection systems to compare different combinations of wild-type probes and mutant probes to obtain consistent detection results. The specific probe combinations are shown in Table 4. The detection probes are coated with microspheres (detection There is a 30T spacer sequence between the probe and the microsphere), the amplification primer (the 5' end of the biotin-labeled reverse primer (SEQ ID NO.6 and SEQ ID NO.7)), the detection method, etc. as implemented Described in Example 1 and Example 2.

[0080] Table 4 Hybridization probe sequence

[0081]

[0082] 2. Sample testing

[0083] Using the liquid phase chip prepared by the above design, samples 21-40 were detected according to the detection ...

Embodiment 4BI

[0091] The selection of embodiment 4BIM gene mutation detection detection probe

[0092] 1. Design of liquid-phase chip preparation (selection of wild-type and mutant-specific primer sequences)

[0093] Using the forward or reverse complementary sequence of the target sequence where the BIM gene deletion mutation is located as a template, design detection probes for wild type and mutant type respectively, including 2 preferred detection probes and 4 preparations in Example 1 of the present invention. The selected detection probes are shown in Table 7. Detection probe-coated microspheres (with a 30T spacer sequence between the detection probe and the microspheres), amplification primers (biotin-labeled reverse primers (SEQ ID NO.6 and SEQ ID NO.7) 5 'end), detection methods, etc. are as described in Example 1 and Example 2.

[0094] Table 7 Hybridization probe sequence

[0095]

[0096]

[0097] 2. Sample testing

[0098] Using the liquid phase chip prepared by the ab...

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Abstract

The present invention discloses a detection liquid phase chip and detection probes for BIM gene deletion mutation. The detection liquid phase chip comprises: BIM gene deletion mutation wild-type targeted detection probe coated microspheres and BIM gene deletion mutation mutant-type targeted detection probe coated microspheres, wherein the wild-type detection probe is SEQ ID NO.1 or SEQ ID NO.2, and the mutant-type detection probe is SEQ ID NO.3 or SEQ ID NO.4; and biotin labeled amplification primers. According to the present invention, the prepared BIM gene mutation detection liquid phase chip has a good signal-to-noise ratio, no cross-reaction basically exist among the designed detection probes, the amplification primers and the amplification product, and parallel detection on the wild-type sequence and the mutant-type sequence in the same reaction system can be achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a BIM gene deletion mutation detection probe and a liquid phase chip. Background technique [0002] The BIM gene exists on human chromosome 2 and is a pro-apoptotic member of the B-cell leukemia / lymphoma 2 (BCL2 gene) protein family. The BIM gene expresses a protein containing the BH3 (BCL2-homology domain3) domain, which can inhibit tumor cell growth by combining with some growth-promoting proteins of the Bcl2 family Bcl2-l1, BCL-XL, BCL2A1, MCL1, or promote apoptosis The combination of protein BAX and BAK1 promotes tumor cell apoptosis. The BH3 domain is the key functional domain of the BIM gene, and studies have found that this functional domain is closely related to the function of TKIs. For example, the normal expression of BIM gene is required in non-small cell lung cancer (NSCLC) treated with TKIs drug gefitinib, and BH3-like drugs ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6876C12Q2600/106C12Q2600/156
Inventor 许嘉森刘志明
Owner SUREXAM BIO TECH
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