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Cervical cancer diagnosis and prognosis kit

A diagnostic kit and the technology of the kit, which are applied in the field of biomedicine, can solve the problems of artificial influence of cytology detection, low specificity of HPV detection, and high probability of missed diagnosis, and achieve strong primer specificity, easy to explore and improve amplification conditions. The effect of detection accuracy

Active Publication Date: 2021-02-19
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main method of clinical cervical cancer screening is cervical liquid-based cytology combined with HPV virus detection, but the cytology detection is greatly influenced by human beings, the probability of missed diagnosis is high, the specificity of HPV detection is low, and there is overtreatment limitations

Method used

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  • Cervical cancer diagnosis and prognosis kit
  • Cervical cancer diagnosis and prognosis kit
  • Cervical cancer diagnosis and prognosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. MSC-related information obtained from genome-wide methylation microarray screening

[0056] Collect 10 cases of normal cervical tissue, CIN3 tissue and cervical cancer tissue, and ensure that the content of epithelial cells (normal cervix) or neoplastic cells (CIN3 or cervical cancer) in the slices is >30% ( figure 1 ), using the Illumina Infinium MethylationEPIC Bead Chip genome-wide methylation chip to screen 850k methylation sites, and perform differential DNA methylation sites according to the method reported in Comprehensive analysis of DNA methylation data with RnBeads (Nat Methods.2014). Click Filter.

[0057] 2. Verification in patients with normal cervical epithelium-precancerous lesions-cancer

[0058] 1. QMSP Verification

[0059] The paraffin specimens of normal cervix-precancerous lesions-cancer patients were collected in the Obstetrics and Gynecology Department of Qilu Hospital, including 36 cases of normal cervical tissue, 36 cases of CIN and 85 case...

Embodiment 2

[0079] Embodiment 2 A kind of cervical cancer diagnostic kit

[0080] 1. The kit used is:

[0081] (1) DNA extraction of paraffin tissue specimens: QIAamp DNA FFPE Tissue (Qiagen, Germany) kit

[0082] (2) DNA methylation modification: EZ DNA Methylation-Gold TM (Zymo Research, USA) Kit

[0083] (3) Methylated and unmethylated DNA standards: EpiTect PCR Control DNA Set (Qiagen, Germany)

[0084] (4) Real-time PCR reaction kit: Power SYBR Green PCR Master Mix (ABI, USA) kit

[0085] (5) PCR reaction kit: AmpliTag Gold 360 Master Mix (ABI, USA) kit

[0086] 2. Real-time PCR conditions:

[0087] Real-time PCR reaction conditions:

[0088]

[0089] Real-time PCR result judgment:

[0090] The formula for calculating the degree of MSC methylation is 2 [Ct(β-actin)-Ct(target)] ×10,000;

[0091] If the CT value of the internal reference β-actin is greater than 32, it is considered that the sample does not meet the quality control and the sample is unqualified;

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Abstract

The invention particularly relates to a cervical cancer diagnosis and prognosis kit. Cervical cancer is still a big threat to female health at present, and at the present stage, a cervical cancer screening mode is cervical liquid-based cytological examination combined with HPV (human papillomavirus) detection, but a cytological examination result is greatly influenced by human subjectivity, and the accuracy of the detection result is insufficient. Therefore, the kit has important significance in providing a more accurate cervical cancer early diagnosis technology. Based on the current situation, the positive correlation between the MSC methylation degree and the cervical cancer development condition is confirmed through screening work, and based on the discovery, the kit for judging earlyscreening, diagnosis and prognosis condition of cervical cancer is provided, detection is carried out in a QMSP mode, and the detection sensitivity and specificity are remarkably improved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a kit for early screening, diagnosis and prognosis judgment of cervical cancer. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Cervical cancer is the fourth most common cancer in women and the fourth leading cause of cancer death in women. According to the latest global data released by the International Agency for Research on Cancer, there were 569,847 new cases of cervical cancer worldwide in 2018, and 311,365 people died of cervical cancer. Although HPV vaccination and early cervical cancer screening have become effective strategies to prevent the disease,...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/118C12Q2531/113C12Q2545/101C12Q2561/113
Inventor 焦薪霖张青崔保霞孔北华
Owner SHANDONG UNIV QILU HOSPITAL
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