Kit and method for methylation detection of cervical high-grade cervical lesions and cervical cancer related genes

Abstract

The invention discloses a kit and method for methylation detection of cervical lesions and cervical cancer related gene. The kit contains reagents including a PCR reaction solution, eight primers fixed to a PCR octa-tube, a positive reference substance, a negative reference substance and a cell lysate for extracting a sample DNA. The detection method comprises the steps of extracting the sample DNA, processing and purifying the sample DNA, preparing a PCR reaction system and adding the sample, carrying out a PCR amplification reaction, collecting a fluorescence signal, obtaining and analyzingan amplification curve and a melting curve, obtaining a Ct value and a Tm value of each marker by analyzing the amplification curve and the melting curve, formulating a positive determination method based on a weighting algorithm, carrying out positive determination on each marker and integrating, finally explaining sample results. The kit has the advantages of high sensitivity, strong specificityand non-invasiveness, and is particularly suitable for auxiliary diagnosis of cervical high-grade lesions or cervical cancer and HPV-positive shunt management.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a kit and method for detecting methylation of cervical high-grade lesions and cervical cancer-related genes. Background technique [0002] Cervical cancer is one of the common female malignant tumors, its incidence is second only to breast cancer, ranking second in gynecological tumors. According to the latest data released by the National Cancer Registry Center, there were 98,900 new cervical cancer cases and 30,500 deaths in my country in 2015, and the incidence and mortality rates are increasing year by year. Scientific research has confirmed that persistent infection of high-risk papillomavirus (hrHPV) is the main cause of cervical cancer, and HPV-negative women have an extremely low risk of cervical cancer. However, most women who test positive for HPV do not develop precancer or cancer, and only a minority of women with HPV infection are at risk of developi...

AI Technical Summary

Problems solved by technology

Among them, the missed diagnosis rate of VIA is high, and the specificity of cytology is high, but the sensitivity is low, and the establishment of cytology depends on local health resources, and it is difficult to establish in low-resource areas.

The sensitivity of HPV DNA detection is relatively high, but most HPVs are transient infections and will not develop into cervical precancerous lesions or cervical cancer, causing unnecessary panic to women and wasting medical resources

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