RT-PCR primers for detecting transcriptional level of GRP78 mRNA in serum as well as kit and method for evaluating concurrent intestinal cancer susceptibility of hyperglycemia population

A RT-PCR, transcription-level technology, applied in DNA/RNA fragments, biochemical equipment and methods, recombinant DNA technology, etc., can solve the problems of application limitations, inability to accurately quantify mRNA, easy contamination and false positive rate, etc.

Inactive Publication Date: 2015-05-20
HUZHOU NO 1 PEOPLES HOSPITAL
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Problems solved by technology

However, in the application of traditional PCR technology, there are shortcomings such as inability to accurately quantify specific mRNA, and easy contamination during operation, resulting in a high false positive rate, so it cannot meet the needs of actual work and its application is limited.

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  • RT-PCR primers for detecting transcriptional level of GRP78 mRNA in serum as well as kit and method for evaluating concurrent intestinal cancer susceptibility of hyperglycemia population
  • RT-PCR primers for detecting transcriptional level of GRP78 mRNA in serum as well as kit and method for evaluating concurrent intestinal cancer susceptibility of hyperglycemia population
  • RT-PCR primers for detecting transcriptional level of GRP78 mRNA in serum as well as kit and method for evaluating concurrent intestinal cancer susceptibility of hyperglycemia population

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Embodiment Construction

[0075] Below in conjunction with embodiment the present invention is further described. However, it should be clear that the above description and the following examples herein are only for illustration, and do not constitute a limitation to the scope of the present invention.

[0076] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0077] The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

[0078] 1. Primer Design and Synthesis

[0079] 1.1 Design and synthesis of GRP78 gene primers

[0080] The standard sequence of the GRP78 gene to be amplified is 1282-1456 bases in the exon 6 region of the GRP78 gene, and primers are designed using the full-length mRNA sequence of GRP78 as a template. The F sequence of the upstream primer of the standard PCR is: 5'- CCCGTCCAGAAAGTGTTG -3'; the R sequence of the downstream primer is: 5'- CAGCACCATACGCT...

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Abstract

The invention relates to an RT-PCR kit and a method for evaluating the concurrent intestinal cancer susceptibility of a hyperglycemia population by using the kit, belonging to the technical field of real-time fluorescence quantitative reverse transcription polymerase chain reaction kit development and human pathology combination. RT-PCR primers for detecting the transcriptional level of GRP78 mRNA in serum include a primer F: 5'-CCCGTCCAGAAAGTGTTG-3' and a primer R: 5'-CAGCACCATACGCTACAG-3'. The invention establishes a method for detecting GRP78 mRNA in peripheral vein serum by using the specific primers; and a quantitative PCR amplification technology is adopted in the method, so that the sensitivity for detecting GRP78 mRNA is greatly improved, and enough information can be obtained in extremely few specimens.

Description

technical field [0001] The present invention relates to a primer, a kit and a method for evaluating the susceptibility of hyperglycemia crowd complicated with colorectal cancer; in particular, it relates to a RT-PCR primer for detecting GRP78 mRNA transcription level in serum, which is used for assessing hyperglycemia crowd complicated with colorectal cancer RT-PCR kit for susceptibility and method for assessing susceptibility to colorectal cancer in hyperglycemia population. It belongs to the technical field of the combination of real-time fluorescent quantitative reverse transcription polymerase chain reaction kit development and human pathology. Background technique [0002] According to the statistics of the World Health Organization (WHO), more than 220 million people in the world suffer from diabetes, and 3.4 million people died of severe complications caused by high blood sugar in 2004. By 2030, the number of deaths due to diabetes will double again . Among the Chin...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/118C12Q2600/158
Inventor 冯文明崔戈鲍鹰郭慧慧薛涛王翔
Owner HUZHOU NO 1 PEOPLES HOSPITAL
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