Ovarian cancer susceptibility-related molecular marker as well as detection primer and kit

A technology of molecular markers and susceptibility, which is applied in the fields of medicine and molecular biology, can solve problems such as the correlation of calcium-sensing receptors, and achieve the effect of convenient and specific amplification, easy amplification conditions, and strong specificity

Active Publication Date: 2015-10-07
SHANDONG UNIV QILU HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no report on the correlation betwe

Method used

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  • Ovarian cancer susceptibility-related molecular marker as well as detection primer and kit
  • Ovarian cancer susceptibility-related molecular marker as well as detection primer and kit
  • Ovarian cancer susceptibility-related molecular marker as well as detection primer and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Detection of CASR Gene Mutation and Association Analysis with Ovarian Cancer

[0027] 1.1 Research object

[0028] A total of 290 patients with ovarian cancer were selected and diagnosed and treated in Qilu Hospital of Shandong University from September 2008 to October 2014. The clinical data were obtained from medical records. 312 age-matched control patients were healthy women who underwent physical examination in the Health Examination Center of Qilu Hospital. Most of the subjects were Han residents in Shandong Province. Take 1.5ml of peripheral blood from the test personnel and store it in a freezer at -80°C. All subjects obtained consent in accordance with the requirements of the Ethics Committee of Qilu Hospital of Shandong University.

[0029] 1.2 DNA extraction

[0030] The peripheral blood DNA of the subjects was extracted by the conventional phenol-chloroform method, as follows:

[0031] (1) Take 400ul of blood and add it to a 1.5ml centrifuge t...

Embodiment 2

[0055] Ovarian Cancer Susceptibility Detection Kit

[0056] Since the mutation 719A>G in SEQ ID NO: 1 is highly correlated with the publication of ovarian cancer, CASR gene-specific primers can be designed based on this mutation and then expanded detection using the patient's DNA as a template.

[0057] Prepare a test kit (100 person-times), which contains the substances shown in Table 3:

[0058] Table 3 Composition of the kit

[0059]

[0060] 2ml of peripheral blood was drawn from the subjects, and DNA was extracted by conventional methods. Ovarian cancer susceptibility detection kit was used for PCR reaction, the reaction products were sequenced, and the sequencing results were checked and analyzed using Meglign 7.0 and Chromas 2.33 software. The susceptibility of subjects with rs17251221 (719A>G) in the test results was lower than that of normal people.

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Abstract

The invention discloses an ovarian cancer susceptibility-related molecular marker as well as a detection primer and a kit. The nucleotide sequence of the ovarian cancer susceptibility-related molecular marker is shown by SEQ ID NO:1; and compared with a normal CASR gene, the 719th site has single nucleotide polymorphism: A>G. The primer is shown by SEQ ID NO:2 and SEQ ID NO:3 in the sequence table; and the primer can be used for specifically amplifying the CASR gene, and the length of the amplified fragment is 200-2,700bp. The invention also provides a method for detecting in vitro whether a sample has the single nucleotide polymorphism of the CASR gene. The primer and kit for detecting the ovarian cancer susceptibility-related molecular marker have the advantages of simple, accurate and fast detection and high specificity and are of great importance in effective early diagnosis, early treatment and individual prevention on the high-risk population or susceptible individuals and ovarian cancer patients.

Description

technical field [0001] The present invention relates to a molecular marker related to ovarian cancer susceptibility, and primers and kits for detecting molecular markers related to ovarian cancer susceptibility, in particular to single nucleotide polymorphism (single nucleotide polymorphism, SNP) and its correlation with ovarian cancer, as well as primers and kits for detecting these SNPs, belong to the fields of medicine and molecular biology. Background technique [0002] Ovarian cancer is the most lethal female reproductive system tumor. In 2014, there were 21,980 new cases and 14,270 deaths in the United States. Due to insidious onset and inconspicuous symptoms, most patients are diagnosed at an advanced stage. The 5-year survival rate of patients with advanced ovarian cancer is only about 30-45%, and the prognosis is poor. However, the 5-year survival rate of patients with early ovarian cancer is as high as 90%. It reminds us that the key to improving the prognosis and...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/156
Inventor 孔北华闫实苑存忠杨其峰
Owner SHANDONG UNIV QILU HOSPITAL
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