Method, primers and probe for detecting relative expression quantity of 11q23/MLL fusion genes

Abstract

The invention discloses a method for detecting the relative expression quantity of 4 fusion genes belonging to 11q23 / MLL series, and primers and a probe for detecting the relative expression quantity of 4 fusion genes of 16 fusion types in total belonging to 11q23 / MLL series. Through test, the primers and the probe are good in specificity, high in sensitivity and simple and convenient to operate, the data which is obtained on the basis of double-standard curve method of house-keeping gene and target gene by utilizing a real-time fluorescence quantification PCR method of the Taqman probe can be used for detecting the relative expression level of the fusion genes belonging to 11q23 / MLL series in patients with ALL (Acute Lymphoblastic Leukemia), AML (Acute Myeloblastic Leukemia) and MDS (Myelodysplastic Syndrome), the high-risk groups can be accurately screened, the detection time can be effectively saved, and the detection precision can be enhanced.

Description

technical field [0001] The invention belongs to the field of life science and biotechnology, in particular a primer, a probe and a detection method for detecting the relative expression levels of 16 fusion types in total of 4 fusion genes of 11q23 / MLL series. Background technique [0002] The MLL (mixed lineage leukemia, MLL) gene is located on the long arm of chromosome 11, region 2, band 3 (11q23), and is one of the genes that are often involved in hematologic malignancies. an important factor. So far, many partner genes on chromosome 11q23 have been identified, forming more than 70 fusion genes, at least 50 of which have been cloned at the molecular level. Recent studies have shown that 11q23 / MLL acute leukemia (11q23 / MLL acute leukemia, 11q23 / MLL AL) has different partner genes, leukemia cell types, patient age and treatment strategies, and the prognosis is very different, and these patients have different effects on conventional chemotherapy. Good, the three-year surv...

AI Technical Summary

Problems solved by technology

Among them, 1) The result of traditional chromosome banding technology is intuitive, but it requires good metaphase and chromosome morphology. For duplication, deletion or inversion of small chromosome fragments, microtranslocations cannot be interpreted, and tandem duplication dup11q23 cannot be detected the exception

2) Although fluorescence in situ hybridization (FISH) has high detection sensitivity, it can only detect common chromosomal abnormalities, and is not suitable for rare and unknown abnormalities.

3) Multiplex nested RT-PCR is the most widely used method for detecting various fusion genes today. It can detect a variety of common MLL fusion genes on a large scale, but can only provide qualitative or rough quantitative results (semi-quantitative) , and the sensitivity is limited by the methodology, and there is a possibility of missed detection for the detection of small residues. More importantly, nested RT-PCR has high requirements for the experimental environment, which is prone to serious cross-contamination, resulting in false positive results

Among them, SYBR Green I, due to the use of unsaturated dyes, can produce non-specific binding to any DNA double helix structure, and its specificity must be judged by observing the melting curve, so the specificity is not ideal; the double-probe hybridization method uses two Probes, although very specific, are prohibitively expensive

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