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Kit for real time fluorescence quantitative RT-RCR detection of human cancer embryoantigen

A real-time fluorescence quantification, kit technology, applied in biological testing, fluorescence/phosphorescence, microbial determination/inspection, etc., can solve the problems of inaccurate quantification and poor specificity, and achieve improved detection sensitivity, specificity, and reduction. The effect of false positive rate

Inactive Publication Date: 2003-10-29
安徽普元生物科技股份有限公司
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Problems solved by technology

In the past, qualitative RT-PCR was used to detect CEA mRNA, but from a technical point of view, although qualitative PCR has significantly improved sensitivity compared with traditional ELISA and other methods, its specificity is relatively poor, and it cannot be accurately quantified.

Method used

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  • Kit for real time fluorescence quantitative RT-RCR detection of human cancer embryoantigen
  • Kit for real time fluorescence quantitative RT-RCR detection of human cancer embryoantigen

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Embodiment 1

[0035] Fluorescent quantitative RT-PCR method to detect the expression of CEA and its application

[0036] 1. Materials:

[0037] Trizol reagents and restriction endonucleases were purchased from LTI / Gibco, USA, pGEM-T-Easy cloning system, MMLV reverse transcriptase, Taq DNA polymerase, Oligo(dT) 15-18Purchased from Promega Company in the United States, the sequencing reagents, 377 sequencer, 2400 PCR instrument and 7700 quantitative PCR instrument were all products of Perkin Elmer Company in the United States.

[0038] 2. Primer and probe design and synthesis:

[0039] Using the full-length cDNA sequence of CEA (GenBank accession number M17303) as a template, use Primer Express TM (V1.0, Perkin Elmer, USA) software analyzes TaqMan primers and probe sites, and selects the best combination based on the simultaneous consideration of the CEA genomic DNA sequence.

[0040] Standard PCR upstream primer sequence is: 5′-AGACAGTCACAGCAGCCTTGAC-3′, downstream primer sequence is 5′-G...

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Abstract

A reagent kit for real-time quantitatively fluorescent RT-PCR detection of human cancino-embryonic antigen is composed of reaction liquid, M-MLV reverse transcriptase, Rnasin, PCR reaction liquid, Taq, standards and references. It can precisely and quantitatively detect the mRNA expression of CEA in specimen.

Description

Technical field [0001] The invention belongs to the field of biological technology, and is a kind of cDNA obtained by reverse transcription (RT) mRNA sample, combined with real-time fluorescent quantitative PCR detection technology, can accurately and quantitatively detect the mRNA expression level of human carcinoembryonic antigen (CEA) in the specimen kit. Background technique [0002] The invention belongs to the field of biological technology, and is a kit for obtaining eDNA from a reverse transcription (RT) mRNA sample, combined with real-time fluorescent quantitative PCR detection technology, which can accurately and quantitatively detect the mRNA expression level of CEA in a sample. [0003] Metastasis of malignant tumors is a major problem in clinical treatment and an important cause of death from malignant tumors. Early detection and diagnosis are of great significance to the selection of clinical treatment options for tumors, which can greatly improve the survival...

Claims

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Application Information

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IPC IPC(8): C12Q1/04G01N21/64G01N33/53
Inventor 曹江郑树张行叶景佳
Owner 安徽普元生物科技股份有限公司
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