Kit for detecting expression level of 17 beta-hydroxysteroid dehydrogenase 7 in human prostate cancer

A technology for hydroxysteroids and prostate cancer, applied in the biological field, can solve problems such as low resolution and inaccurate quantification, achieve high sensitivity, reduce false positive rate, and improve detection sensitivity

Inactive Publication Date: 2010-06-30
FUDAN UNIV
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  • Description
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Problems solved by technology

The above methods have extremely high sensitivity, but based on the characteristics of the PCR reaction, the product amount at the end of the reaction often cannot correctly reflect the DNA c

Method used

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  • Kit for detecting expression level of 17 beta-hydroxysteroid dehydrogenase 7 in human prostate cancer
  • Kit for detecting expression level of 17 beta-hydroxysteroid dehydrogenase 7 in human prostate cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Fluorescent quantitative RT-PCR detection of 17β-HSD7 expression in human prostate cancer samples

[0036] 1. Experimental materials

[0037] Trizol reagent was purchased from Invitrogen Company of the United States; MMLV reverse transcriptase, Taq DNA polymerase, and OligodT were purchased from Promega Company of the United States; 7900HT fluorescent quantitative PCR instrument was purchased from ABI Company of the United States.

[0038] 2. Primer and probe design and synthesis:

[0039] 17β-HSD7, GAPDH full-length cDNA sequence as a template, using Primer Express TM 2.0 software analyzes TaqMan primers and probe sites, and selects the best combination.

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Abstract

The invention belongs to the field of biotechnology and relates to a real-time fluorescent quantitative PCR kit for detecting the mRNA expression level of 17 beta-hydroxysteroid dehydrogenase 7 (17 beta-HSD7) in human prostate cancer samples. The kit contains 2 * reverse transcription reaction solution, 20 * reverse transcriptase, water treated by diethyl pyrocarbonate, 2 * polymerase chain reaction solution, 20 * primers and fluorescent probes for detection and a standard product. The invention establishes a method for utilizing the Taqman technology for detecting the expression of the 17 beta-HSD7, and the detection practices prove that the kit and the detection method are feasible. As the method adopts the PCR amplification technology, the method can greatly improve the detection sensitivity of the 17 beta-HSD7 and obtain enough information in a very small number of samples; due to the application of the fluorescent probes, the specificity is also greatly improved and the false positive rate of the conventional PCR amplification is reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a real-time fluorescent quantitative PCR kit for detecting the expression of 17β-hydroxysteroid dehydrogenase 7 (17β-HSD7) mRNA in human prostate cancer samples. Background technique [0002] Prostate cancer is one of the most common malignant tumors in men. According to the data, there are obvious regional differences in the incidence and mortality of prostate cancer. The United States, Northern Europe, Western Europe and Australia are high-incidence areas of prostate cancer, while Asia and North Africa are relatively low-incidence areas. However, the incidence of prostate cancer in areas with low incidence of prostate cancer has also increased significantly since the 1970s. In the past, the incidence of prostate cancer in my country was relatively low, but with the prolongation of life expectancy, the change of dietary structure and the advancement of diagnostic technology, its in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
Inventor 姜梅李瑶谢毅毛裕民
Owner FUDAN UNIV
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