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In-situ hybridization testing kit of ALDH1 gene and testing method and application thereof

A detection kit and in situ hybridization technology, applied in the field of kits, can solve the problem that ALDH1 gene detection technology has not been reported, and achieve the effects of convenient operation, strong specificity and high sensitivity

Inactive Publication Date: 2011-03-30
SUZHOU FUYING GENE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report about the in situ hybridization detection kit and detection technology of ALDH1 gene

Method used

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  • In-situ hybridization testing kit of ALDH1 gene and testing method and application thereof
  • In-situ hybridization testing kit of ALDH1 gene and testing method and application thereof
  • In-situ hybridization testing kit of ALDH1 gene and testing method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] An in situ hybridization detection kit for ALDH1 gene, comprising a hybridization probe, a marker, and a potentiator, wherein the sequence of the hybridization probe is shown in SEQ ID NO.1. Hybridization probes were labeled with digoxigenin. The composition of other liquids and specimens in the kit is as follows:

[0045] Digestive solution 100μl / tube 1 tube / box Colorless transparent liquid

[0046] Protective solution 100μl / tube 1 tube / box Colorless transparent liquid

[0047] Pre-hybridization solution 1300μl / tube 2 tubes / box Colorless transparent liquid

[0048] Sense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0049] Antisense hybridization solution 10μl / tube 1 tube / box Colorless transparent liquid

[0050] Blocking solution 1000μl / tube 1 tube / box Colorless transparent liquid

[0051] Alkaline phosphatase antibody 1μl / tube 1 tube / box Colorless transparent liquid

[0052] Chromogen A 175μl / tube 1 tube / box Yellow liquid

[0053] ...

Embodiment 2

[0095] A kind of ALDH1 gene in situ hybridization detection method and its kit application

[0096] 1. Specimen processing

[0097] 1. Use a 10ml centrifuge tube to fill 4.5ml of lymphocyte separation medium, then slowly add 3ml of anticoagulated blood into the centrifuge tube containing lymphocyte separation medium (blood: lymphocyte separation medium = 1:1.5), and centrifuge at 2000r / min 10min;

[0098] 2. Take the white blood cells in the middle layer into another centrifuge tube, then add about twice the amount of 1× buffer I to this tube, mix well, and centrifuge at 1500g / min for 10min;

[0099] 3. Discard the supernatant. Add about twice the 1× buffer I to the precipitate, mix well, and centrifuge at 1500g / min for 10min;

[0100] 4. Discard the supernatant, and absorb the excess liquid from the mouth of the test tube with paper towels. Then the precipitate was made into a suspension, dropped on a glass slide, and allowed to dry naturally. (Hospitals with conditions c...

Embodiment 3

[0134] Parallel experiments between the detection of breast cancer disease with the ALDH1 gene kit and the detection of breast cancer disease with the ERK gene kit.

[0135] In order to scientifically evaluate the specificity, sensitivity and accuracy of the above genes in breast cancer. We use the method of parallel experiment, detect the mRNA of above-mentioned gene at the same time, detection technology adopts nucleic acid in situ hybridization technology, use the peripheral blood of the same breast cancer patient, detect the mRNA of ALDH1 gene and ERK gene at the same time (carry out nucleic acid in situ hybridization, Immunohistochemical staining, microscopic counting, result reporting, etc. all used the same methods, steps and reagents as the in situ hybridization technique in Example 1 and Example 2). It was found that the expression level of ALDH1 gene in patients with breast cancer was higher than that of ERK gene in patients with the same disease. The results showed...

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Abstract

The invention relates to an in-situ hybridization testing kit of an aldehyde dehydrogenase (ALDH1) gene. The kit comprises a hybridization probe and a marker, wherein the sequence of the hybridization probe is shown as SEQ ID No. 1. The invention also provides an in-situ hybridization testing method of the ALDH1gene. In addition, the invention also provides application of the kit to preparation of medicaments for testing breast cancer or prognostic diseases of breast cancer. The kit provided by the invention has the characteristics of high flexibility and specificity. The testing method of the invention is convenient and easy to operate and can be generally used and popularized in district-level or city-level hospitals.

Description

【Technical field】 [0001] The invention relates to a kit, in particular to an in situ hybridization detection kit of ALDH1 gene and its detection method and application. 【Background technique】 [0002] According to the information provided by authoritative organizations at home and abroad, there are 1.6 million new cancer cases in my country every year, nearly 1.6 million deaths, and 6 million patients. Globally, there are 8 million new cancer patients every year, nearly 8 million deaths, and about 84 million patients. , by 2020 the number of people will double, which is a set of terrible figures. Breast cancer is a malignant tumor that seriously endangers women's health. In recent years, the incidence rate has steadily increased and tends to be younger. The incidence rate ranks third among female tumors. According to worldwide statistics, more than 1.2 million people around the world suffer from breast cancer every year, and 200,000 people die from breast cancer every year. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 张云福裘建英
Owner SUZHOU FUYING GENE TECH
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