Kit for detecting blood circulation tumor cells

A technology of tumor cells and peripheral blood circulation, which is applied in the application field of detecting the level of circulating tumor cells in patients with lung cancer. The effect of improving efficiency

Inactive Publication Date: 2015-07-15
刘志东 +2
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Problems solved by technology

[0002] At present, the detection of circulating tumor cells in lung cancer patients mostly adopts the immunomagnetic bead sorting method. Although this method has high accuracy, its sensi...

Method used

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Embodiment Construction

[0052] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with specific examples.

[0053] The detection process of epithelial cell adhesion molecule expression includes the following 5 steps:

[0054] 1. Take 2 mL of peripheral blood from the median cubital vein of patients with non-small cell lung cancer, store the blood in EDTA anticoagulant blood vessels, and store it in a 4°C refrigerator;

[0055] 2. Centrifuge at 300G for 10 minutes, discard the upper layer of plasma, and keep the blood cell layer;

[0056] 3. Use the erythrocyte lysate to remove the red blood cells in the blood cells and retain the nucleated cells;

[0057] 4. Add the above-treated samples to Trizol solution to extract total RNA, and obtain the corresponding cDNA samples through reverse transcription reaction;

[0058] 5. The qRT-PCR absolute quantitative method was used to det...

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Abstract

The technical scheme of the invention discloses a method for performing epithelial cell adhesion molecule expression detection on peripheral blood of lung cancer patients by the utilization of combining an MGB probe with a fluorescent quantitative polymerase chain reaction method, and therefore a circulating tumor cell level in the body is reflected. The method of the kit for detecting blood circulation tumor cells comprises the steps that RNA extraction and cDNA reverse transcription are performed on an objective specimen needing to be detected, then the MGB probe is used to perform fluorescent quantitative polymerase chain amplification reaction, finally the fluorescence intensity is detected by a fluorescent quantitative polymerase chain amplification reaction apparatus, and the epithelial cell adhesion molecule expression level is obtained. The kit for detecting the blood circulation tumor cells has the advantages of being high in sensitivity and high in specificity, meanwhile, the high through-put advantage of a common polymerase chain reaction method is inherited, the detection efficiency is improved, and the detection cost is reduced.

Description

technical field [0001] The invention relates to detecting the expression of epithelial cell adhesion molecules by using MGB probe combined with fluorescent quantitative polymerase chain reaction method to obtain the expression level of epithelial adhesion molecules, and the application of the test method in detecting the level of circulating tumor cells in lung cancer patients. Background technique [0002] At present, the detection of circulating tumor cells in lung cancer patients mostly adopts the immunomagnetic bead sorting method. Although this method has high accuracy, its sensitivity is insufficient. Facing the limitation of decreasing sample size, this method is not enough for mass Disease screening of cancer patients and multiple postoperative efficacy evaluations. [0003] EpCAM (epithelial cell adhesion molecule) was developed by Herlyn et al in 1979 [1] Found in colon cancer, it is considered to be the main tumor-associated antigen in human colon cancer tissue. ...

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 刘志东李云松苏崇玉
Owner 刘志东
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