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Composition and diagnostic reagent for detecting high-grade cervical lesions and cervical cancer

A diagnostic reagent and composition technology, applied in the field of internal reference gene primers, can solve problems such as insufficient correlation of cervical cancer, achieve simple and accurate risk classification methods, and improve the effect of predictive ability

Active Publication Date: 2021-04-13
苏州凯爱健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the problem of lack of correlation with high-grade cervical lesions and cervical cancer existing in the existing methylation detection, the present invention increases The HPV16L1 gene 6367, 6389 sites and HPV18L2 gene 4256, 4261, 4265, 4269, 4275, and 4282 sites methylation levels, which are also closely related to cervical cancer, can be detected to achieve more accurate and effective cervical high-grade lesions and cervical cancer. The technology of cancer auxiliary diagnosis, triage management and prognosis judgment can improve the efficiency of cervical cancer screening

Method used

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  • Composition and diagnostic reagent for detecting high-grade cervical lesions and cervical cancer
  • Composition and diagnostic reagent for detecting high-grade cervical lesions and cervical cancer
  • Composition and diagnostic reagent for detecting high-grade cervical lesions and cervical cancer

Examples

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Embodiment 1

[0057] Example 1 Preparation of diagnostic reagents for cervical high-grade lesions and cervical cancer detection according to the present invention 1. Design and synthesis of primers and probes

[0058] For human EPB41L3 425, 427, 438 sites and the internal reference gene ACTB (for the sequence, refer to the human genome sequence NC_00018.10 published by the NCBI database), and for the viral HPV 16L1 gene 6367, 6389 sites (for the sequence, see AF125673.1 published by the NCBI database) and the HPV18L2 gene Sites 4256, 4261, 4265, 4269, 4275, and 4282 (for the sequence, see AY262282.1 published in the NCBI database), specific primers and probes were designed using Primer Premier 5.0 and Methy Primer Express v1.0 software, respectively.

[0059] Specific primers and probe sequences are shown in the table below:

[0060] sequence name oligonucleotide sequence base length EPB41L3 forward primer TAAATTGGCGGAGAAGGGAC 20 EPB41L3 reverse primer GGTTTTTTT...

Embodiment 2

[0081] Example 2: Detection of cervical cancer methylation using the reagent of Example 1

[0082] 1. Technical principle

[0083] After extracting and purifying the DNA of cervical exfoliated cells, the DNA needs to be treated with bisulfite. After the treatment, the C base on the CpG island that has undergone methylation in the DNA remains as a C base without methylation. The C base on the converted CpG island becomes a U base, which causes the sequence difference between methylated DNA and unmethylated DNA, which can be used to detect methylation for subsequent methylation detection methods level, such as figure 1 shown.

[0084] The methylated target genes EPB41L3, HPV16L1 and HPV18L2 were detected by methylation-specific fluorescent quantitative PCR method. Firstly, methylation-specific primers were used to amplify the nucleic acid, and multi-channel probe fluorescent labeling was used to realize the detection of 3 Real-time fluorescence detection was carried out for e...

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Abstract

The invention discloses a composition and diagnostic reagent for detecting high-grade cervical lesions and cervical cancer. The composition comprises specific primers for detecting 425, 427 and 438 site methylation of a human EPB41L3 gene, 6367 and 6389 site methylation of a virus HPV 16L1 gene and 4256, 4261, 4265, 4269, 4275 and 4282 site methylation of an HPV18L2 gene, and respective reference primers. According to the composition and diagnostic reagent for detecting the high-grade cervical lesions and cervical cancer, by combining human EPB41L3 and virus HPV 16L1 and HPV18L2 methylation detection, the high-risk HPV virus methylation level serving as the direct pathogenesis of cervical cancer is brought into a detection system for the first time, the sensitivity is high, the specificity is high, and the prediction accuracy of the high-grade cervical lesions and cervical cancer is remarkably improved.

Description

technical field [0001] The present invention belongs to the technical field of biomedicine, and specifically relates to a composition and a diagnostic reagent or a kit for detecting high-grade cervical lesions, specifically, specific primers for methylation of EPB41L3, HPV16L1 and HPV18L2 and their respective internal reference genes primers. Background technique [0002] The morbidity and mortality of cervical cancer ranks second among female malignancies. According to the statistics of WHO in 2018, there are 589,800 new cases of cervical cancer in the world. In my country, there are about 130,000 to 150,000 new cases every year, accounting for about 30% of the world. In 2018, the number of deaths from cervical cancer in my country was about 47,000 to 48,000, and the incidence and mortality rates were on the rise year by year. Although cervical cancer is a malignant tumor, normal cervical cells develop into invasive cancer through precancerous lesions, and there may be a ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6886C12Q2600/118C12Q2600/154C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 施晨阳阿蒂拉·洛林茨石立立祝旭景陈华韩亮
Owner 苏州凯爱健康科技有限公司
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