Primer probe and kit for detecting oncogene of cervical cancer and detection method for non-detection disease purpose

A technology of oncogenes and detection methods, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc.

Active Publication Date: 2013-05-22
ZHENGZHOU KODIA BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there are no relevant reports on the detection of cervical cancer oncogenes using bDNA technology, and the design and specificity of probes have important practical significance

Method used

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  • Primer probe and kit for detecting oncogene of cervical cancer and detection method for non-detection disease purpose
  • Primer probe and kit for detecting oncogene of cervical cancer and detection method for non-detection disease purpose
  • Primer probe and kit for detecting oncogene of cervical cancer and detection method for non-detection disease purpose

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0152] HeLa cells were used to design three gradients, the original gradient was 100cells / μL, the secondary gradient was 50cells / μL, and the final gradient was 25cells / μL. All three gradients have the same volume. Schematic diagram of 96-microwell plate simulation layout figure 1 shown.

[0153] Sampling: Before sampling, wipe the mucus and blood on the surface of the cervix with a cotton swab, and gently insert the bristle part of the front of the sampler deep into the channel of the cervix, so that the bristles at the back can fully touch the transition of the cervix, and gently move forward Hold the sampler and turn the sampler 3-5 full circles in the same clockwise direction.

[0154] Sample processing: set the incubator to the temperature required for lysis at 65°C; match the number of the centrifuge tube and the sample to confirm that it is correct; mix the liquid in the sample tube and transfer it to the centrifuge tube, centrifuge at 3000rpm for 5min, pour off the su...

Embodiment 2

[0191] Sampling: Before sampling, wipe the mucus and blood on the surface of the cervix with a cotton swab, and gently insert the bristle part of the front of the sampler deep into the channel of the cervix, so that the bristles at the back can fully touch the transition of the cervix, and gently move forward Hold the sampler and turn the sampler 3-5 full circles in the same clockwise direction.

[0192] Sample processing: set the incubator to the temperature required for lysis at 65°C; match the number of the centrifuge tube and the sample to confirm that it is correct; mix the liquid in the sample tube and transfer it to the centrifuge tube, centrifuge at 3000rpm for 5min, pour off the supernatant, and add 2ml to the centrifuge tube. Ionized water, suspended sediment, 3000rpm, 5min centrifugation, discard the supernatant; add 200μL lysate, 400μL deionized water, 5μL proteinase K to each sample, mix well, put it in an incubator at 65°C for 1 hour, and mix it once in a while. ...

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Abstract

The invention relates to the technical field of biologics, and in particular relates to a primer probe and kit for detecting an oncogene of a cervical cancer and a detection method for a non-detection disease purpose. The probe is high in specificity; and the false positive rate is reduced. By the method, quantified detection for nucleic acid to be detected is realized by hybridization, signal magnification and enzymatic chemiluminescence; the magnification factor is determined without a magnification process for increasing an index; and the repetitiveness and the stability are high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer probe for detecting cervical cancer oncogenes, a kit and a detection method for non-detection of diseases. Background technique [0002] Cervical cancer is the third most common malignant tumor in women in the world after breast cancer and colorectal cancer, and the second most common malignant tumor in developing countries after breast cancer. It is the most common cancer of the female reproductive tract. Malignant tumor. In 2008, there were estimated to be 529,800 new cervical cancer cases and 255,100 deaths worldwide, of which 85% were in developing countries (Jemal, 2011). With the development of cervical cancer screening, the incidence and mortality of cervical cancer in developed countries have decreased significantly. [0003] Cervical cancer is currently the only gynecological malignancy with a clear etiology, which is associated with persistent infection of high-r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 张鹭鹭张爱国刘桐宇
Owner ZHENGZHOU KODIA BIOTECHNOLOGY CO LTD
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