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HER-2 gene and / or TOP2A gene detection probe and preparation method and kit

A HER-2 and gene detection technology, which is applied in the direction of DNA preparation, biochemical equipment and methods, and microbial determination/inspection, can solve the problems of lack of specificity and high detection kits, achieve high sensitivity and reduce hybridization background , good effect of specificity

Inactive Publication Date: 2016-04-27
GUANGZHOU LBP MEDICINE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there is still a lack of specific detection kits for FISH detection of HER-2 gene and / or TOP2A gene

Method used

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  • HER-2 gene and / or TOP2A gene detection probe and preparation method and kit
  • HER-2 gene and / or TOP2A gene detection probe and preparation method and kit
  • HER-2 gene and / or TOP2A gene detection probe and preparation method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation of embodiment 1HER-2 gene and TOP2A gene detection probe

[0041] A preparation method for HER-2 detection probe (GSPHER-2), comprising the following steps:

[0042] Clone screening: select clones containing the target gene HER-2 and both ends of the sequence, such as Figure 1A shown.

[0043] GSPHER-2 includes two groups, and the following two groups of detection probes are prepared respectively.

[0044] HER-2chr17:37,856,254-37,884,915,28,662bp

[0045]

[0046] The preparation method of described TOP2A detection probe (GSPTOP2A), comprises the following steps:

[0047] Clone screening: select clones containing the target gene TOP2A and both ends of the sequence, such as Figure 1B shown.

[0048] GSPTOP2A includes two groups, as shown in the following table, which were purchased from Invitrogen RP11BAC and CTDBAC clone libraries. The following two groups of detection probes were prepared respectively.

[0049] TOP2Achr17:38,544,773-38,574,2...

Embodiment 2

[0063] Example 2: Preparation method of HER-2 gene and TOP2A gene detection kit

[0064] HER-2 gene and TOP2A gene detection kit include two components of HER-2 / TOP2A hybridization fluid and DAPI counterstaining agent, wherein HER-2 and TOP2A hybridization fluid comprise a set of GSPTOP2A gene probes described in Example 1 and a set of GSPHER-2 gene probes. HER-2 and TOP2A gene have two groups of detection probes respectively, two groups of two kinds of genes can be combined at will, in the present embodiment, described HER-2 gene and TOP2A gene detection kit are four kinds, are respectively: HER-2 gene and TOP2A gene detection kit 2 (group 1) + TOP2A (group 1) combination, HER-2 (group 2) + TOP2A (group 2) combination, HER-2 (group 1) + TOP2A (group 2) combination, HER-2 (group 2) Group 2) + TOP2A (group 1) combination, CSP17 probe (chromosome 17 identification probe, purchased from D17Z1 SpectrumAquaProbe, Cat. No.: 06J38-017), buffer components for hybridization environmen...

Embodiment 3

[0073] Example 3: Detection method of HER-2 gene and TOP2A gene detection kit

[0074] 1. Slide pretreatment

[0075] 1.1 Place the slides in a 65±5°C incubator and bake overnight;

[0076] 1.2 Take out the slide and put it in xylene to dewax at room temperature for 15 minutes;

[0077] 1.3 Take out the slide, and put it into another vat of xylene to continue dewaxing at room temperature for 15 minutes;

[0078] 1.4 Take out the slide, and put it in absolute ethanol at room temperature for 10 minutes to remove residual xylene;

[0079] 1.5 Take out the slide, and put it into 100%, 90%, 70% graded ethanol for rehydration at room temperature for 3 minutes each;

[0080] 1.6 Take out the slide, put it in purified water and wash it at room temperature for 3 minutes, and absorb excess water with a lint-free paper towel;

[0081] 1.7 Take out the slides, put them into purified water and boil them at 100±5°C for 25 minutes (place the slides horizontally in the container with the ...

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Abstract

The present invention relates to a HER-2 gene and / or TOP2A gene detection probe and a preparation method thereof, and the method comprises the following steps: selecting BAC aiming at HER-2 gene to clone into RP11-1021P11, RP11-62N23 and RP11-1044P23, or selecting BAC to clone into RP11-98J2, RP11-1065L22 and CTD-2327I15; and / or selecting BAC aiming at TOP2A gene to clone into RP11-1152A10 and CTD-3217C5, or selecting BAC to clone into RP11-737D6, CTD-3087O22, RP11-48O10 and CTD-2134K5; obtaining plasmid DNA; and marking. The present invention also discloses a kit which includes the HER-2 gene and / or TOP2A gene detection probe. The best HER-2 gene and TOP2A gene detection probe is screened, so that a signal counting line is accurate and rapid, and results are well reproducible.

Description

technical field [0001] The invention belongs to biotechnology, and in particular relates to a HER-2 (ERBB2) gene and / or TOP2A gene detection probe, a preparation method and a kit. Background technique [0002] HER-2 (ERBB2) gene has gene amplification and protein overexpression in 20%-30% of breast cancer patients. HER-2 is currently recognized as an important prognostic / predictive factor for breast cancer. In multivariate analysis results, HER-2 is an independent prognostic factor for tumor recurrence and overall survival. HER-2 positive breast cancer has strong invasiveness, short disease-free survival and poor prognosis. Trastuzumab (Herceptin) is suitable for breast cancer with HER-2 overexpression and gene amplification. TOP2A gene regulates the supercoiled state of DNA, directly participates in cellular processes, and is the target enzyme of anthracycline antibiotics. Patients with TOP2A gene abnormalities have a poor prognosis and shorter recurrence-free survival, e...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12N15/10C12N15/11C12Q1/68C12Q1/6886C12Q2600/106C12Q2600/166
Inventor 陈绍宇何瑰席影
Owner GUANGZHOU LBP MEDICINE SCI & TECH
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