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Quality control sample for detecting breast cancer and preparation method of quality control sample

A quality control product, breast cancer technology, applied in the fields of clinical laboratory, pathology and biology, can solve the problems of inability to specifically reflect HER2 false positives, limited sources of quality control products, poor consistency, etc. Simple operation, repeatable and consistent results

Active Publication Date: 2015-04-29
BEIJING HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditionally, quality control products tend to use tissue samples from breast cancer patients as detection quality control products, but the source of such quality control products is limited, the consistency is poor, and there are great limitations in the application process
In addition, the patient-derived quality control can only be randomly determined to be HER2 positive or negative, and cannot specifically reflect the expression level of the HER2 gene and the false positives caused by polysomy

Method used

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  • Quality control sample for detecting breast cancer and preparation method of quality control sample
  • Quality control sample for detecting breast cancer and preparation method of quality control sample
  • Quality control sample for detecting breast cancer and preparation method of quality control sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: PCR method to verify the expression of HER2 gene in various tumor cell lines

[0047] The breast cancer cell line was cultured to the logarithmic growth phase according to the method in step 1 of Example 2, and the following experiments were carried out using the kit QIAampDNA Mini Kit (Qiajie Enterprise Management Co., Ltd., China).

[0048] 1. DNA extraction:

[0049] ①The three breast cancer cell lines cultured to the logarithmic phase were absorbed into 1×10 6 cells into a centrifuge tube.

[0050] ② Turn on the metal bath at 56°C; shake and mix AL (lysate) before use, and heat in a 56°C water bath if there is precipitation.

[0051] ③Treat the three types of cells obtained above with 200ul PBS, and mix well by pipetting.

[0052] ④Add 20μl QIAGEN Proteinase K to it; suck out the above 220ul sample into a 1.5ml EP tube, add 200ul Buffer AL (lysate), and shake with a vortex shaker for 15 seconds to confirm the formation of a homogeneous state.

[0053]...

Embodiment 2

[0078] Example 2: Preparation of Quality Control Products for FISH Detection of Breast Cancer Cell Line HER2 Gene Amplification

[0079] 1. Culture breast cancer cell lines:

[0080] The MCF-7 cell line was cultured with DMEM medium containing 10% fetal bovine serum, the medium contained 10% fetal bovine serum, 100IU / ml penicillin and 100IU / ml streptomycin; the MDA-MB-453 cell line was cultured with 20% Fetal bovine serum cultured in DMEM medium containing 20% ​​fetal bovine serum, 100IU / ml penicillin and 100IU / ml streptomycin; SKBR-3 cell line cultured with 20% fetal bovine serum McCoy's 5a medium Contains 20% fetal bovine serum, 100IU / ml penicillin and 100IU / ml streptomycin. Cultured in a constant temperature cell incubator at 37°C containing 5% CO2, digested with 0.25% trypsin for cell passage, and expanded the three cell lines to 10 7 -10 8 order of magnitude and in the logarithmic growth phase.

[0081] 2. Preparation of multisomatic cell lines:

[0082] The MCF-7 ce...

Embodiment 3

[0091] Embodiment 3: Observation of the cell distribution of the quality control substance by HE staining

[0092] 1. Method:

[0093] 1. Sample treatment: preheat the quality control substance for 10 minutes before staining, dewax in xylene for 5 minutes; replace with fresh xylene, and dewax for 5 minutes; absolute ethanol for 5 minutes; 90% ethanol for 2 minutes; 70% ethanol for 2 minutes; ethanol for 2 minutes; distilled water for 2 minutes.

[0094] 2. Staining: staining with hematoxylin staining solution for 5-10 minutes; differentiation in HCL aqueous solution for 5 seconds; soaking in tap water for about 5 minutes; washing again with distilled water (several seconds); 95% ethanol for 5 seconds. ; Staining with eosin staining solution for 30 seconds to 2 minutes.

[0095] 3. Dehydration and transparent sealing: dehydration with 95% ethanol for 2 minutes; replace with fresh 95% ethanol for another 2 minutes; xylene for 5 minutes; replace with fresh xylene for another 5 ...

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Abstract

The invention discloses a quality control sample for detecting breast cancer and a preparation method of the quality control sample, particularly relates to a quality control material for HER2 gene amplification FISH detection and a preparation method of the quality control material, and belongs to the technical fields of clinical laboratory medicine, pathology and biotechnology. The quality control sample for detecting the breast cancer is a paraffin-embedded specimen slice prepared by a cultivated breast cancer cell line. The quality control sample has the advantages that the quality control sample prepared by the breast cancer cell line cultured in vitro can be used as substitute goods of the quality control sample for fabricating real tissue samples, and is high in sample capacity and high in repeatability; and detection of different HER2 gene levels can be finished by selecting difference cell lines. The quality control sample is simple and convenient in preparation method, and easy to store, can be put into volume production, and can be applied to internal quality control and external quality assessment.

Description

technical field [0001] The invention relates to a breast cancer detection quality control substance and a preparation method thereof, in particular to a quality control substance for HER2 gene amplification FISH detection and a preparation method thereof, belonging to the fields of clinical laboratory science, pathology and biotechnology. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, and its incidence is increasing year by year. Proto-oncogene HER2 (human epidermal growth factor receptor 2 gene) is one of the hotspots of current research. According to data, about 20-30% of invasive ductal carcinomas of breast cancer have overexpression of HER2 gene. A large number of studies have confirmed that HER2 gene amplification is not only closely related to the occurrence, development and prognosis of breast cancer, but also crucial to the selection of drug treatment options. [0003] HER2-positive breast cancer has poor response t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57415
Inventor 李金明李禹龙张瑞韩彦熙王露楠张括孙宇林贵高谢洁红
Owner BEIJING HOSPITAL
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