Typing detection kit for three subtypes of BCR-ABL1 fusion gene

A BCR-ABL1 and gene detection technology, applied in the field of genes, can solve the problems of inconsistent amplification efficiency of internal control and detection of subtypes, low reverse transcription efficiency, etc., and achieve the effect of reducing pollution and waste and ensuring consistency

Pending Publication Date: 2020-11-03
PILOT GENE TECH HANGZHOU CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The invention discloses a detection kit for detecting three subtypes of BCR-ABL1 fusion gene, which solves the problems of low reverse transcription efficiency, external internal control, and often inconsistent amplification efficiency of detection subtypes in the prior art. question

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  • Typing detection kit for three subtypes of BCR-ABL1 fusion gene
  • Typing detection kit for three subtypes of BCR-ABL1 fusion gene
  • Typing detection kit for three subtypes of BCR-ABL1 fusion gene

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Embodiment 1

[0043] like figure 1 As shown, this embodiment discloses a method for simultaneously detecting three subtypes of the BCR-ABL1 fusion gene, including:

[0044]1. According to the sequence of the BCR-ABL1 fusion gene, primers and probes were designed using primer express software. The specific information is shown in Table 1 below:

[0045] Table 1

[0046]

[0047] 2. Samples

[0048] (1) P190 / P210 RNA samples: from clinical samples, RNA was extracted using an RNA extraction kit.

[0049] (2) P230 DNA: DNA fragments synthesized by biological companies.

[0050] (3) ABL1 gene wild-type sample: RNA of oral exfoliated cells was extracted.

[0051] 3. The reverse transcription system is shown in Table 2 below:

[0052] Table 2

[0053]

[0054]

[0055] 4. Reverse transcription conditions

[0056] 1h at 42°C, 15min at 70°C.

[0057] 5. The cDNA digital PCR amplification system is shown in Table 3 below:

[0058] table 3

[0059] Reagent Volume (μL) ...

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Abstract

The invention belongs to the field of genes, and particularly relates to a typing detection kit for three subtypes of a BCR-ABL1 fusion gene. Firstly, the invention discloses a primer probe composition for detecting three subtype genes of BCR-ABL1 at the same time, the three subtype genes of BCR-ABL1 are respectively P190, P210 and P230, and the detected nucleotide sequences are shown as SEQ ID NO: 1-9. A specific downstream primer of the ABL1 gene is adopted for reverse transcription, and the reverse transcription efficiency is higher than that of a random primer and oligo dT; three subtypesand internal control are qualitatively and quantitatively detected by multiple systems at the same time, so that pollution and waste possibly caused by sample adding for multiple times are reduced; amultiplex amplification system adopts the same downstream primer, so that the consistency of amplification efficiency is ensured as possible; and an internal control probe is placed in an amplicon, sothat the abnormal condition that the fusion ratio exceeds 100% is avoided.

Description

technical field [0001] The invention belongs to the field of genes, in particular to a BCR-ABL1 fusion gene three subtype typing detection kit. Background technique [0002] Acute lymphocytic leukemia (ALL) is a common hematological malignancy with high morbidity and mortality, which seriously endangers the physical and mental health of the population (especially adolescents). The etiology and pathogenesis of the vast majority of ALL are not yet fully understood. At present, it is generally believed that it is the result of the interaction between genetic factors and environmental factors. Among the genetic factors, the occurrence of fusion genes is considered to be one of the most important risk factors. As early as 1960, Nowwell and Hungerford discovered an abnormal "miniature chromosome" in the chromosome G of seven chronic myeloid leukemia (CML) cases. This newly discovered abnormal chromosome was later named the Philadelphia chromosome (Philadelphia chromosome, PhChro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851C12N15/11
CPCC12Q1/6886C12Q1/6851C12Q2600/158C12Q2600/16C12Q2531/113C12Q2537/143C12Q2521/107
Inventor 朱海涛夏江
Owner PILOT GENE TECH HANGZHOU CO LTD
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