Application of amz1 gene in preparation of diagnostic kit

A diagnostic kit and gene technology, applied in the field of biomedicine, can solve problems such as inconsistent analysis results

Inactive Publication Date: 2016-04-27
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, differences in different experimental platforms and samples lead to many inconsistencies in the analysis results

Method used

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  • Application of amz1 gene in preparation of diagnostic kit
  • Application of amz1 gene in preparation of diagnostic kit
  • Application of amz1 gene in preparation of diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening for genes associated with colon cancer

[0044] 1.1 NCBIGEO (GeneExpressionOmnibus) data retrieval

[0045] The GEO (Gene Expression Omnibus) database is developed and maintained by NCBI (National Center for Biotechnology Information). The GEO database is the largest database of gene expression data. Gene expression serial analysis) data, SARST (ribosomal sequence tag continuous analysis) data, MS (mass spectrometry) data, proteome data and next-generation high-throughput sequencing data (MPSS, massively parallel sequencing technology), etc.

[0046] a. Search keywords: ("colorectalcancer"[MeSHTerms]OR"colorectalcancer"

[0047] [AllFields]) AND "Homosapiens" [porgn]

[0048] b. Sample selection strategy in the study:

[0049] Limit the research type to "expression profiling by array" Data sets that meet the following criteria will be included in our research: ①The selected data sets must be genome-wide expression mRNA transcriptome data, and all ...

Embodiment 2

[0054] Embodiment 2QPCR verifies the relationship between candidate genes and colon cancer

[0055] Based on the results of meta-analysis of the colon cancer gene chip data, according to the size of the Pvalue, we selected the AMZ1 gene (Meta analysis showed that the expression of the AMZ1 gene was down-regulated in colon cancer tissues compared with normal tissues) for verification. Collect 50 colon cancer tissue samples and 50 normal tissue samples at the same time, and use fluorescent real-time quantitative PCR to carry out classical molecular biology experiment verification (qRT-PCR). The specific operation steps are as follows:

[0056] (1) RNA extraction

[0057] After collecting the samples, freeze them in liquid nitrogen. After taking them out, put the tissue into a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0058] ① Add Trizol and store at room temperature for 5 minutes;

[0059] ② Add 0.2ml of chloroform, vibrate the centrifuge tube vigo...

Embodiment 3

[0068] The preparation of embodiment 3 colon cancer diagnosis kits

[0069] According to the correlation between the AMZ1 gene and colon cancer, whether colon cancer occurs can be diagnosed by detecting the expression of the AMZ1 gene. Accordingly, the present invention provides a test kit for diagnosing colon cancer based on the detection of AMZ1 gene expression. The diagnostic kit The components are as follows: SYBRGreen polymerase chain reaction system; primer pair for amplifying AMZ1 gene and GAPDH gene. The forward primer sequence for amplifying AMZ1 gene is 5'-AAGAACAACAAGCCAGGGGACG-3', the reverse primer sequence is 5'-CTGGAAGGAACTTGCTGAAGGTGA-3'; the forward primer sequence for amplifying GAPDH is 5'-TTTAACTCTGGTAAAGTGGATAT-3', reverse The primer sequence was 5'-GGTGGAATCATATTGGAACA-3'. The SYBRGreen polymerase chain reaction system includes PCR buffer, dNTPs, and SYBRGreen fluorescent dye. The PCR buffer composition is: 25mMKCL, 2.5mMMgCL 2 , 200mM (NH 4 ) 2 SO ...

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Abstract

The invention discloses an application of an AMZ1 gene to preparation of a kit for diagnosing colorectal cancers (CRCs). Five CRC cases and expression chip data of genome mRNA (messenger ribonucleic acid) of normal people, which meet the requirements, are brought into the range of study through data retrieval by utilizing a GEO (gene expression omnibus) of the NCBI (National Center for Biotechnology Information). The AMZ1 gene is screened to be a gene with significant difference by carrying out meta analysis on the chip data. The expression of the AMZ1 gene, compared with normal tissues, in the CRC tissues is reduced, and the conclusion is verified by using a florescent real-time quantitative PCR (polymerase chain reaction) method. Accordingly, the invention also discloses the kit for diagnosing the CRCs and a method for diagnosing the CRCs by utilizing the kit. According to the invention, on the one hand, a new idea for studying the molecular mechanisms of the CRCs is provided, and on the other hand, the more sensitive diagnostic kit is provided for early diagnosis of the CRCs.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a colon cancer diagnostic kit and the application of AMZ1 gene in preparing the colon cancer diagnostic kit. Background technique [0002] The earliest and most widely used gene chip technology in transcriptome research is to extract RNA from the sample to be tested, and use fluorescently labeled nucleotides to reverse transcribe it into cDNA. The labeled nucleotide sequence can be compared with The probe hybridization on the specific site of the gene chip, and the cell gene expression information can be obtained by detecting the hybridization signal. Gene chip technology has become a very stable and credible experimental technology, and a large amount of transcriptome data published so far is mainly generated by gene chip technology. [0003] Colon cancer is the third most common malignant tumor in both men and women. According to the 2013 American Cancer Statistics Report, there were 1...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/158
Inventor 董伟伟屈雪玲王硕李小梅赵慧霞肖文华
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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