ERG gene detection probe, preparation method thereof and reagent kit

A gene detection and kit technology, applied in the field of ERG gene detection probes and its preparation, can solve the problems of meeting the domestic market demand and promoting clinical application, and achieve the goals of adjuvant treatment plan selection, good repeatability, and increased signal brightness Effect

Inactive Publication Date: 2016-03-23
GUANGZHOU LBP MEDICINE SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These problems are extremely unfavorable for meeting domestic market demand and promoting clinical application

Method used

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  • ERG gene detection probe, preparation method thereof and reagent kit
  • ERG gene detection probe, preparation method thereof and reagent kit
  • ERG gene detection probe, preparation method thereof and reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] The preparation of embodiment 1 ERG gene detection probe

[0033] The preparation method of the described ERG detection probe of this implementation comprises the following steps:

[0034] Select the clones containing the target gene ERG and both ends of the sequence, such as figure 1 shown. GSPERG includes two sets of probes:

[0035] The first group of probes includes the first probe and the second probe, as shown in the following table, which were purchased from Invitrogen RP11BAC and CTDBAC clone libraries.

[0036] The second group of probes includes the third probe and the fourth probe, as shown in the table below, which were purchased from Invitrogen RP11BAC and CTDBAC clone libraries.

[0037] The following two groups of detection probes were prepared respectively.

[0038] first set of probes

[0039]

[0040] Second set of probes

[0041]

BAC

Insert segment start and end position

third probe

RP11-360N24

chr21:4001391...

Embodiment 2

[0052] Embodiment 2: ERG gene detection kit preparation method

[0053] The ERG gene rearrangement detection kit includes two components of a hybridization solution and a DAPI counterstain, wherein the hybridization solution contains two groups of GSPERG gene probes (two-color) described in Example 1, and is used for the hybridization environment (to promote hybridization). Buffer components, COTHumanDNA with blocked repeats, etc. DAPI counterstaining agent is mainly used for cell counterstaining after hybridization, in which DAPI will bind to DNA, making the nucleus show blue fluorescence, and the counterstaining agent containing p-phenylenediamine can keep the fluorescence stable.

[0054] The specific formula is as follows:

[0055] (1) Preparation of hybridization solution

[0056]

[0057]

[0058] (2) DAPI counterstain preparation

[0059] Dissolve 10 mg of p-phenylenediamine in 1 ml of PBS, adjust the pH to 9.0, add 9 ml of glycerin, shake and mix repeatedly, a...

Embodiment 3

[0062] Embodiment 3: the detection method of ERG gene detection kit

[0063] 1. Slide pretreatment

[0064] 1.1 Place the slides in a 65±5°C incubator and bake overnight;

[0065] 1.2 Take out the slide and place it in xylene (or environmental dewaxing agent) at room temperature for 10 minutes;

[0066] 1.3 Take out the slide, and put it in another cylinder of room temperature xylene (or environmental dewaxing agent) for 10 minutes;

[0067] 1.4 Take out the slide, and put it in absolute ethanol at room temperature for 10 minutes to remove residual xylene (or environmental dewaxing agent);

[0068] 1.5 Take out the slide, and put it into 100%, 90%, 70% graded ethanol for rehydration at room temperature for 3 minutes each;

[0069] 1.6 Take out the slide, put it in sterilized purified water and wash it at room temperature for 3 minutes, and absorb excess water with a lint-free paper towel;

[0070] 1.7 Take out the slide, put the slide into 1×EDTA repair solution, and repai...

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Abstract

The invention relates to an ERG gene detection probe and a preparation method thereof. The method comprises the following steps that two sets of BAC clones are selected, wherein one set is at least one of RP11-720N21 and CTD-2556C4, and the other set is at least one of RP11-360N24 and RP11-110N12; plasmids are extracted from the clones to obtain plasmid DNA, and quantitation is performed; fluorescein is used for labeling. The invention further discloses a reagent kit containing the ERG gene detection probe. The optimal ERG detection probe is obtained through screening, the signal counting row is accurate is rapid, and the result repeatability is good; the defect of clinical ERG mutation detection is overcome, more patients benefiting from targeted drugs are screened out, the patient survival rate and overall survival are improved.

Description

technical field [0001] The invention belongs to biotechnology, and in particular relates to an ERG gene detection probe, a preparation method and a kit thereof. Background technique [0002] Prostate cancer is a common malignant tumor disease in elderly men. In European and American countries, the death rate of prostate cancer ranks second among all kinds of malignant tumor diseases. The incidence of prostate cancer in my country is much lower than that in western countries; however, due to changes in diet, environment, and living habits, the incidence has increased significantly in recent years. The staging characteristics and survival rate of prostate cancer in China show that most of the patients who see a doctor are advanced prostate cancer patients, and the treatment effect is not good. Therefore, early detection of prostate cancer has become an important topic. The fusion of TMPRSS2 gene and ETS generally exists only in prostate cancer, and does not appear in other ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6841C12Q1/6886C12Q2600/106C12Q2600/156C12Q2563/107C12Q2563/173C12Q2537/163C12N15/11C12Q1/68
Inventor 何瑰陈绍宇张会清
Owner GUANGZHOU LBP MEDICINE SCI & TECH
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