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Method and kit for quickly detecting expression quantity of CK19 mRNA (messenger ribonucleic acid)

A technology of expression amount and kit, which is applied in the detection field of CK19mRNA expression amount, can solve the problems of short detection time, sensitivity and specificity to be further studied, and achieve the effects of fast detection speed, improved sensitivity and repeatability, and simple operation

Active Publication Date: 2013-12-11
北京明谛生物医药科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has a short detection time and can better exclude interference from human DNA or CK19 pseudogenes, but its sensitivity and specificity need further study

Method used

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  • Method and kit for quickly detecting expression quantity of CK19 mRNA (messenger ribonucleic acid)
  • Method and kit for quickly detecting expression quantity of CK19 mRNA (messenger ribonucleic acid)
  • Method and kit for quickly detecting expression quantity of CK19 mRNA (messenger ribonucleic acid)

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Experimental program
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Embodiment

[0062] 1. Materials

[0063] 1. Experimental samples: sentinel lymph node tissue from four breast cancer patients, from a hospital in Beijing;

[0064] 2. Total RNA extraction kit: RNeasy Mini Kit was purchased from QIAGEN (Cat.74104);

[0065] 3. One-step multiplex qRT-PCR kit: Rotor-Gene Multiplex RT-PCR Kit was purchased from QIAGEN (Cat.204972);

[0066] 4. The first primer and first probe for detecting the expression level of CK19mRNA, the sequence is as follows, synthesized and labeled by QIAGEN;

[0067] The first forward primer: 5'-CCATTGAGAACTCCAGGATT-3' (SEQ ID No.1)

[0068] The first reverse primer: 5'-CCCTGCGCAGGCCGTTGATGT-3' (SEQ ID No.2)

[0069] First probe: 5'-FAM-ACAATGCCCGTCTGGCTGCAGA-BHQ1-3' (SEQ ID No.3)

[0070] 5. The second primer and the second probe used to detect the expression level of the internal reference gene ACTB mRNA, the sequence is as follows, synthesized and labeled by QIAGEN;

[0071] Second forward primer: 5'-TCACCCACACTGTGCCCATCTA-3...

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Abstract

The invention discloses a primer, probe, detection method and kit for detecting expression quantity of CK19 mRNA (messenger ribonucleic acid). The primer comprises a forward primer of which the base sequence is shown as SEQ ID NO:1 and a reverse primer of which the base sequence is shown as SEQ ID NO:2; the base sequence of the probe is shown as SEQ ID NO:3; and the 5' terminal is marked with a reporting fluorophore, and the 3' terminal is marked with a quenching fluorophore. The method disclosed by the invention has the advantages of high detection speed, simple operation process, high specificity, high sensitivity and high reproducibility, can quickly and specifically detect the expression quantity of CK19 mRNA in a detection sample, can be used for correlation detection using CK19 as a marker, and has wide application range and favorable application prospects.

Description

technical field [0001] The invention relates to the detection of CK19mRNA expression, in particular to primers, probes, detection methods and kits for rapid detection of CK19mRNA expression. Background technique [0002] Cytokeratin (CK) is the main skeleton protein in keratinocytes. The main function of this structural protein is to maintain the integrity and continuity of epithelial tissue. Studies have found that cytokeratins are highly conserved and specific for tissue differentiation, and are closely related to the proliferation and differentiation of epithelial cells. There are currently 20 types of cytokeratins that have been confirmed, namely CK1-20. Cytokeratin is a commonly used tumor immunohistochemical marker, and its positive expression is more common in epithelial cells, mesothelial cells, carcinoma, and mesothelioma. [0003] Cytokeratin 19 (CK19) is one of the cytokeratin components distributed in epithelial cells. It is a soluble acidic polypeptide molecule...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 王殊刘建云
Owner 北京明谛生物医药科技有限公司