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Kit for detecting circulating tumor cells

A technology of tumor cells and kits, applied in the field of detection, can solve the problems of relying on subjective judgment, poor sensitivity and specificity, etc., and achieve the effect of high positive detection rate, high detection sensitivity and specificity

Inactive Publication Date: 2016-08-24
郝淮杰
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The methods currently used for tumor diagnosis can be classified into three categories: pathology (biopsy sections), imaging (ultrasound, X-ray, CT, PET, etc.) and serology (serum tumor-associated proteins, such as CA-125, CA- 199, CEA, etc.), but these methods have inevitable shortcomings, such as poor sensitivity and specificity, and rely too much on the subjective judgment of doctors, etc.

Method used

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  • Kit for detecting circulating tumor cells
  • Kit for detecting circulating tumor cells
  • Kit for detecting circulating tumor cells

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Experimental program
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Effect test

Embodiment 1

[0016] Sensitivity and specificity evaluation of embodiment 1 real-time quantitative RT-PCR method

[0017] 1. Experimental materials

[0018] 1. Cells: breast cancer cell lines, SK-BR-3, MCF-7, ZR 75-1, MDA-MB-453; human myeloid leukemia cells HL-60, human Burkitt's lymphoma cells Raji, human peripheral blood Leukemic T cells Jurkat were purchased from Peking Union Medical College Cell Resource Center.

[0019] 2. Reagents and instruments: TRIzol reagent, glycogen, EDTA / trypsin cell digestion solution, Ambion TubroDNA-free kit, SuperScript III First-strand synthesis for RT-PCR, Taqman Universal Master Mix II kit, MicroAmp 8-tubes and tubes Covers were purchased from Life Technologies. Primers and probes (Table 1) were synthesized in Invitrogen. StepOne Plus real-time quantitative PCR instrument, Countess TM Automated cell counter, NanoDrop 2000.

[0020] Table 1. Primer and Probe Sequences

[0021]

[0022] 2. Experimental method

[0023] 1. Preparation of total cel...

Embodiment 2

[0042] Example 2 Application of real-time quantitative RT-PCR method in the detection of circulating tumor cells in breast cancer patients

[0043] 1. Experimental materials

[0044] 50ml and 15ml centrifuge tubes were purchased from Corning Company. Ficoll-Paque Plus was purchased from GE Healthcare. BD Vacutainer blood collection tubes (sodium citrate anticoagulant) were purchased from Becton-Dickinson Company. Dynabeads CD45, TRIzol reagent, glycogen, EDTA / trypsin cell digestion solution, Ambion TubroDNA-free kit, SuperScript III First-strand synthesis for RT-PCR, Taqman Universal Master Mix II kit, MicroAmp 8-tubes and caps were purchased from Life Technologies Inc. Primers and probes (Table 1) were synthesized in Invitrogen. StepOne Plus real-time quantitative PCR instrument, Countess TM Automated cell counter, NanoDrop 2000.

[0045] 2. Experimental method

[0046] 1. Preparation of total cellular RNA

[0047] The peripheral blood of healthy volunteers or breast ...

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Abstract

The invention discloses a kit for detecting circulating tumor cells and a method thereof. The kit comprises a realtime quantitative PCR primer, a probe, and experiment materials for cell total RNA extraction, cell cDNA preparation, and realtime quantitative PCR reactions. The invention also discloses a method using the kit to detect tumor cells. The method comprises steps of cell total RNA preparation, cDNA preparation, and realtime quantitative PCR reactions. Through the method, circulating tumor cells in peripheral blood of tumor patients can be identified. The method using the kit can detect circulating tumor cells, which express ERBB2 and / or KRT19, of solid tumors such as breast cancer, lung cancer, colorectal cancer, and the like.

Description

technical field [0001] The invention relates to a kit for detecting circulating tumor cells, in particular to a kit for detecting circulating tumor cells through RT-PCT technology, and also relates to the detection method used. Background technique [0002] The number of cancer patients worldwide is increasing at an alarming rate due to factors such as environmental pollution and people maintaining various unhealthy living habits such as smoking and eating unhealthy food. In 2012, the number of new cancer cases in the world was estimated to be about 14 million. The International Agency for Research on Cancer under the World Health Organization recently published the "2014 World Cancer Report" and pointed out that this number is expected to rise to 22 million within 20 years. "Early prevention, early detection, early treatment" is the best way to prevent and treat cancer. The methods currently used for tumor diagnosis can be classified into three categories: pathology (biops...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 郝淮杰
Owner 郝淮杰
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