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Kit for cervical cancer screening and use method thereof

A kit and technology for cervical cancer, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc., can solve problems that are not specifically pointed out, have low reliability, and have no obvious correlation with cervical cancer, etc. question

Active Publication Date: 2021-05-18
江苏达伯药业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that the abnormal expression of P16INK4a gene is significantly related to the occurrence of cervical cancer and the prognosis of treatment, but there are also reports that the correlation between P16INK4a protein and the occurrence of cervical cancer is not obvious, but is more related to the prognosis
Therefore, it is controversial to use p16INK4a protein alone as a diagnostic and screening index for cervical cancer, and its reliability is not high
[0005] CN 110643706 A patent proposes that Notch1, P16INK4a, and hTERC genes are used as markers for cervical cancer screening, but does not specify how to use these markers, and these markers are all genes for detecting target proteins

Method used

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  • Kit for cervical cancer screening and use method thereof
  • Kit for cervical cancer screening and use method thereof
  • Kit for cervical cancer screening and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Detection of expression levels of miR-210 and LncRNA BLACAT1 in cervical cancer

[0037] Participate in the specific operation process of Example 2 (such as primers, miRNA extraction and fluorescent quantitative RT-PCR reaction) to the expression levels of miR-210 and LncRNA BLACAT1 in 30 cervical cancer patient tissues and adjacent tissues (2-△△CT value) for detection and analysis. For specific results, see figure 1 . It can be seen from the figure that the expressions of miR-210 and LncRNA BLACAT1 are significantly upregulated in cervical cancer.

Embodiment 2

[0038] Example 2 Establishment of the kit for joint detection of the expression levels of miR-210, LncRNABLACAT1 and P16INK4a

[0039] 1. Design primers and Taqman-MGB probes

[0040]According to the miR-210, LncRNA BLACAT1 and P16INK4a sequences, Primer Primer5.0 software was used to design primers and probes. Multiple pairs of specific primers and probes were obtained, and after comparison and screening, a set of optimal primers and MGB probes were finally determined. See the table below for details:

[0041]

[0042]

[0043] Compared with traditional TaqMan-TAMRA probes, TaqMan-MGB probes have the following advantages: (1) Increase TM value: an average of 15 bases can increase 18°C, which can shorten the length of the probe, especially for sequence design with high AT content helps a lot, and improves the difference in TM values ​​between paired and unpaired templates. (2) Improve the signal-to-noise ratio: because the quenching group at the 3' end of the probe is...

Embodiment 3

[0056] Embodiment 3 clinical sample detection

[0057] Use the kit of Example 2 to detect 100 clinical samples, and compare the detection results with the clinical cytopathology. The results show that the coincidence rate of the kit is 99 / 100=99%, and the negative coincidence rate is 61 / 62=98.4% (there is a sample whose pathological test is negative, and the test kit is positive), and the positive coincidence rate is 38 / 38=100%.

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Abstract

The invention discloses a kit for cervical cancer screening. The kit comprises reagents for jointly detecting the expression levels of miR-210, LncRNA BLACAT1 and P16INK4a. The kit can be used as one of means for cervical cancer screening, has the advantages of being rapid and convenient to detect, high in detection sensitivity, good in specificity, low in cost, capable of meeting the detection requirements of most tumor patients and wide in application range.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a test kit for cervical cancer screening and an application method thereof. Background technique [0002] Cervical cancer is the most common gynecological malignancy. The high-incidence age of carcinoma in situ is 30-35 years old, and that of invasive carcinoma is 45-55 years old. In recent years, the onset tends to be younger. Persistent high-risk HPV (human papilloma virus, HPV, human papillomavirus) infection is the main risk factor for cervical cancer, and more than 90% of cervical cancers are associated with high-risk HPV infection. [0003] Human papillomavirus is a papillomavacuolating virus A genus belonging to the Papovaviridae family. It is a non-enveloped double-stranded circular DNA virus consisting of 7900 base pairs. The gene structure of each type of HPV basically includes three important regions: early region (E), late region (Late region, L) and long co...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/158C12Q2600/166C12Q2600/178C12Q2531/113C12Q2545/101C12Q2561/101C12Q2563/107
Inventor 黄小容
Owner 江苏达伯药业有限公司