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Kit used for detecting nasopharyngeal carcinoma and application thereof

A kit and technology for nasopharyngeal carcinoma, applied in the field of kits for detection of methylation in the promoter region of FMR1 gene by fluorescence quantitative method, can solve the problems of Epstein-Barr virus sensitivity, low specificity and limited effect

Active Publication Date: 2019-04-12
敬善生物科技江苏有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epstein-Barr virus infection is a high-risk factor for nasopharyngeal carcinoma, but the sensitivity and specificity of Epstein-Barr virus-related examinations in clinical practice are low, and their role in the early diagnosis of nasopharyngeal carcinoma is limited. Therefore, there is an urgent need for nasopharyngeal carcinoma in clinical practice. A New Approach to Early Diagnosis

Method used

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  • Kit used for detecting nasopharyngeal carcinoma and application thereof
  • Kit used for detecting nasopharyngeal carcinoma and application thereof
  • Kit used for detecting nasopharyngeal carcinoma and application thereof

Examples

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Embodiment 1

[0018] Embodiment one invention detects the kit of nasopharyngeal carcinoma

[0019] The kit for detecting nasopharyngeal carcinoma of the present invention includes reagents for detecting methylation of MSF1 gene and RASSF1A gene based on fluorescent quantitative PCR method and reagents for detecting internal control reference gene ACTB. The reagents for detecting the methylation of MSF1 gene and RASSF1A gene each include a pair of methyl 4-site amplification primers and a probe for detecting methylation sites; the detection of ACTB gene includes a pair of non-methylation site amplification primers , a probe for detecting unmethylated sites. Various sequences are shown in Table 1 and Table 2.

[0020] 1. PCR primer design

[0021] Both MSF1 and RASSF1A gene methylation amplification primer sequences contain CpG sequence (CG sequence in the sequence), and try to make the CpG sequence be located at the 5' end of the primer, so as to ensure that only the methylation amplificat...

Embodiment 2

[0049] Embodiment two. the application of kit of the present invention

[0050] Using the MSF1 gene and RASSF1A gene methylation detection kit and method described in the present invention, based on the ABI7500Fast Dx fluorescence quantitative instrument of ThermoFisher Company, the nasopharyngeal swab samples that have been confirmed as nasopharyngeal carcinoma patients clinically and pathologically, and non- The methylation values ​​of the nasopharyngeal swab samples of the nasopharyngeal carcinoma control population were detected, and the results are shown in Table 3 and Table 4.

[0051] Table 3. Ct values ​​of MSF1 and RASSF1 gene methylation detected in nasopharyngeal swab samples of the nasopharyngeal carcinoma group, and Ct values ​​of the internal reference gene ACTB. When the Ct value > 40, it was considered negative.

[0052]

[0053] Table 4. The methylation of MSF1 and RASSF1 genes detected in nasopharyngeal swab samples of the non-nasopharyngeal carcinoma gro...

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Abstract

The invention relates to a kit used for detecting nasopharyngeal carcinoma and an application thereof. The kit includes a sample DNA extraction reagent, a DNA methylation modification conversion reagent, and a reagent for detecting methylation of MSF1 gene and RASSF1A gene based on a fluorescent quantitative PCR method. Methylation PCR-amplified primer sequences of the MSF1 gene and the RASSF1A gene contain a CpG sequence. The kit simultaneously uses the MSF1 and RASSF1A genes for detecting of nasopharyngeal carcinoma, and the sensitivity and specificity are significantly improved compared with the sensitivity and specificity of the detection by the single use of the MSF1 gene or the RASSF1A gene.

Description

technical field [0001] The invention relates to the field of molecular diagnosis, in particular to a kit for detecting the methylation of the FMR1 gene promoter region by a fluorescent quantitative method and an application thereof. Background technique [0002] Nasopharyngeal carcinoma is a high-incidence tumor in South China, and its treatment is based on radiotherapy. However, for recurrent tumors with advanced stages, especially those with stage III or above, comprehensive treatments such as radiotherapy, chemotherapy, and surgery are not effective. Therefore, early diagnosis is important to improve its clinical treatment effect. Epstein-Barr virus infection is a high-risk factor for nasopharyngeal carcinoma, but the sensitivity and specificity of Epstein-Barr virus-related examinations in clinical practice are low, and their role in the early diagnosis of nasopharyngeal carcinoma is limited. Therefore, there is an urgent need for nasopharyngeal carcinoma in clinical pra...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858
CPCC12Q1/6858C12Q1/6886C12Q2600/16C12Q2600/154C12Q2531/113C12Q2523/125C12Q2537/143
Inventor 李旭辉张雷郭书娟
Owner 敬善生物科技江苏有限公司
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