BCR-ABL fusion gene amplification kit and BCR-ABL fusion gene detection kit

Abstract

The present invention provides a BCR-ABL fusion gene amplification kit and a BCR-ABL fusion gene detection kit, wherein the amplification kit has advantages of convenient use, rapid operation, and high sensitivity. According to the present invention, the BCR-ABL fusion gene can be subjected to typing and quantitation with the detection kit, such that the pathogenetic condition development and the prognosis of the chronic myelocytic leukemia patient can be monitored according to the BCR-ABL fusion gene detection result.

Description

technical field [0001] The invention relates to the field of gene amplification, in particular to a fusion gene amplification kit and detection kit. Background technique [0002] Chronic myelogenous leukemia (CML) is a malignant clonal disease originating from hematopoietic stem cells. It is clinically divided into chronic phase, accelerated phase and blast phase. Acute phase is the main cause of clinical death. Cytogenetic characteristics are Ph Chromosome, namely t(9;22)(q34;q11), forms bcr-abl mRNA at the molecular level. t(9;22)(q34;q11) is a characteristic chromosomal translocation of CML, forming a breakpoint cluster-Abelson oncogene (bcr-abl) fusion gene, where bcr-abl p210 It is the molecular marker of CML. In recent years, foreign studies on the BCR-ABL fusion protein at the molecular level have found that it can be divided into three main types according to the position of the BCR breakpoint: M-bcr, m-bcr, μ-bcr, and the corresponding encoding p210 protein, p190 ...

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Problems solved by technology

[0015] 3. With the continuous development of detection technology, the detection method of BCR-ABL fusion gene is also constantly progressing, from traditional cytogenetics method to fluorescence in situ hybridization method, to molecular level polymerase chain reaction, These methods can detect BCR-ABL transcripts, but various detection methods have their advantages and disadvantages in sample processing and detection operations. In order to obtain better detection results, it is often necessary to use several detection methods in combination

[0017] 1. The detection sensitivity is low, cytogenetics is 5%, FISH is 0.5%, and PCR method can reach 0.001-0.0001%, which is equivalent to 10000-100000 normal cells in one cell. -5 -10 -6 BCR-ABL fusion gene RNA expressed in individual cells

[0018] 2. The disadvantage of poor accuracy. The abnormal detection rate of traditional cytogenetic analysis is only 33.3%. When Ph+<10%, errors are prone to occur, and its sensitivity has been unable to detect minimal residual disease (MRD) in leukemia patients. testing, which limits the utility of cytogenetics in leukemia patients

[0020] 4. The detection cycle is long, the operation is complicated and the use of special instruments. The traditional cytogenetic method generally takes 1-2 weeks for chromosome culture only, and the cells must be cultured to the middle of cell division, which greatly increases the detection time. The culture technology requires high requirements, and the culture is sometimes prone to failure. Even if the culture is successful, some cells will not divide or the number of cell divisions will be insufficient, which will lead to the inability to detect positive results by conventional cytogenetics

The experimental operation of FISH detection is more complicated, it is difficult to detect when the specimen is not handled properly, and false positive results are prone to occur, the cost is high, and certain equipment is required, which makes it difficult to promote the clinical application of FISH

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