A method and kit for detecting mlh1 gene mutation

A kit and gene technology, applied in the fields of molecular biology and medicine, to achieve the effects of clear results, simple and easy detection methods, and reduced burdens

Inactive Publication Date: 2011-12-28
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows individuals who have been identified through their own DNA sequence to make informed decisions about how they want or behave towards developing treatments that target specific genes associated with certain types of tumors such as colonic carcinoma (CRC). By comparing these techniques against patient data obtained during routine medical examinings, this new technique helps clinicians decide which therapies will work best for each individual's needs better than current methods based only on symptoms alone.

Problems solved by technology

The technical problem addressed in this patents relates to identifying specific types or combinations of chromogens associated with certain diseases like necrosis due to radiation exposure, specifically chloroprene poisonings, and gastrointestinal tract nerve injuries. This requires accurate testing and identification of these variants through molecular analysis techniques called sequencing.

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  • A method and kit for detecting mlh1 gene mutation
  • A method and kit for detecting mlh1 gene mutation
  • A method and kit for detecting mlh1 gene mutation

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1 MLH1 gene mutation detection

[0032] 1. Experimental materials:

[0033] (1).Main reagents

[0034] PCR-related reagents: TaKaRa Bao Bioengineering (Dalian) Co., Ltd. high efficiency, including Taq DNA Polymerase for PCR amplification, TaKaRa Code: DRR001A, TaKaRa Ex Taq (5U / μl), 10×Ex Taq Buffer (Mg 2+ Plus), dNTP Mixture (2.5 mM each). Primers: Synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. Marker: Tiangen Biochemical Technology Co., Ltd. (Marker I). Purification kit: Beijing Broadtech Biogene Technology Co., Ltd., B-type small DNA fragment rapid gel recovery kit (100 times).

[0035] (2). Important instruments:

[0036] 1. PCR instrument: Eppendorf AG 22331Hamburg

[0037] eppendorf Mastercycler epgradient S

[0038] 220-240V 50-60Hz 800W

[0039] 2. Centrifuge: Eppendorf AG 22331Hamburg

[0040] eppendorf Centrifuge 5415D

[0041] 230V 0.8A 50-60Hz 180W

[0042] Max speed: 13200min -1 ...

Embodiment 2

[0078] Example 2 Mutation type and pathogenicity analysis results

[0079] The samples were obtained from the Colorectal Cancer Clinic of the Cancer Hospital Affiliated to Fudan University. Among the 10 patients with colorectal cancer who are willing to undergo MLH1 gene testing, the following patients have MLH1 gene mutations, and the specific test results are as follows. All three patients were patients with hereditary nonpolyposis colorectal cancer.

[0080] Table 2

[0081] patient number

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Abstract

The invention belongs to the fields of molecular biology and medicine. The invention provides a method for detecting MLH1 gene mutation: firstly, obtain the DNA sequence or amino acid sequence of the promoter region or coding region of the MLH1 gene in the sample, and then compare it with the corresponding sequence of the normal MLH1 gene to determine whether the MLH1 gene exists in the sample Mutation status; the coding region is the No. 1, No. 2 or No. 19 exon of the MLH1 gene. The invention also provides a corresponding kit for detecting MLH1 gene mutation and the application of the method in determining patients with hereditary nonpolyposis colorectal cancer. The detection method of the invention is simple and easy to implement, low in cost, efficient and fast, and the result is clear. It can greatly reduce the burden on patients, exclude non-HNPCC patients as soon as possible, and provide a reference for the drug regimen of colorectal cancer patients.

Description

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Claims

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Application Information

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Owner FUDAN UNIV
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