BCR-ABL gene fluorescence quantitative RT-PCR primer and probe and reagent kit

A technology of fluorescence quantification and detection kit, applied in the biological field, can solve the problems of unfavorable acute lymphoblastic leukemia clinical classification, disease observation and recurrence monitoring, slow detection speed and low sensitivity

Inactive Publication Date: 2007-07-11
SHANGHAI FOSUN PHARMA (GROUP) CO LTD +1
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AI Technical Summary

Problems solved by technology

[0008] The technical problem to be solved by the present invention is to provide a kind of BCR-ABL fusion gene (P190 BCR / ABL and P210 BCR / ABL ) mRNA fluorescent quantitative RT-PCR primers and probes and kits to overcome the quantitative detection of BCR / ABL fusion gene mRNA expression levels in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL) patient cells Poor accuracy, low sensitivity, and slow detection speed are not conducive to the clinical typing, disease observation and recurrence monitoring of chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL).

Method used

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  • BCR-ABL gene fluorescence quantitative RT-PCR primer and probe and reagent kit

Examples

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Embodiment 1

[0099] Detection of human BCR-ABL fusion gene mRNA expression and its application by real-time fluorescent quantitative RT-PCR

[0100] 1. Primer and probe design and synthesis:

[0101] Using the full-length cDNA sequence of one of the b3a2 subtypes of the BCR-ABL fusion gene (GenBank accession number HSA131466) as a template, the Oligo software was used to analyze the positions of TaqMan primers and probes, and according to the consideration of both BCR and ABL genomic DNA sequences situation, choose the best combination.

[0102] The upstream primer sequence of standard PCR is: 5'-TGT GAA ACT CCA GAC TGT CC-3'; the downstream primer sequence is: 5'-ACG AAA AGG TTG GGG TCA TT-3';

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Abstract

The invention discloses a quantitative RT-PCR primer and probe and agent box of BCR-ABL fusing gene mRNA fluorescence with BCR-ABL fusing gene primer and probe sequence as SEQ ID NO1-4 and internal reference gene primer and probe sequence as SEQ ID NO5-7, wherein the agent box contains cell cracking liquid, water, RT-PCR reacting liquid, internal reference TBP RT-PCR reacting liquid, BCR-ABL fusing gene detecting probe, TBP internal reference gene testing probe, composite enzyme, standard material and comparing material; the box can test the expressive level of mRNA of P210BCR/ABL and P190BCR/ABL in the specimen, which provides the reference to diagnose, recurrent and treat chronic granulocytic leukemia and acute lymphocyte leukemia.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to quantitative reverse transcription PCR detection of gene mRNA expression. Background technique [0002] ABL is a proto-oncogene, located on chromosome 9 q34, the gene product is a non-receptor tyrosine protein kinase, but usually has no kinase activity. The BCR gene is located on chromosome 22 q11, and the normal her gene product is a 160kD cytoplasmic phosphoprotein. Due to the translocation of t(9;22)(q34;q11), the ABL proto-oncogene located on chromosome 9 is juxtaposed with the BCR gene located on chromosome 22, forming a new BCR-ABL fusion gene. In the translocation, the ABL proto-oncogene breakpoint on chromosome 9 was relatively constant, but the BCR gene breakpoint on chromosome 22 was not constant, and most of the breakpoints were located upstream and downstream of exon 14. There are mainly three subtypes of fusion genes formed according to different breakpoints, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 沈维祥吴大治夏懿
Owner SHANGHAI FOSUN PHARMA (GROUP) CO LTD
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