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Kit for detecting human BCR-ABL fusion gene and use method thereof

A technology that integrates genes and kits, and is applied in biochemical equipment and methods, and microbial measurement/inspection, etc. It can solve the problems of lack of Chinese patient data and achieve the effects of improved specificity, high accuracy, and wide linear range

Pending Publication Date: 2019-10-22
苏州云泰生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2002, 26 laboratories from 10 European countries participated in the formulation of the "European Cancer Prevention Program", established a set of standardized fusion gene detection methods for leukemia, and laid a theoretical foundation for large-scale clinical fusion gene detection, but because Lack of data on Chinese patients, failure to apply on a large scale in China

Method used

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  • Kit for detecting human BCR-ABL fusion gene and use method thereof
  • Kit for detecting human BCR-ABL fusion gene and use method thereof
  • Kit for detecting human BCR-ABL fusion gene and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] A test kit (20 tests / part) for detecting human BCR-ABL fusion gene, its composition is as shown in Table 7:

[0109] Table 7:

[0110]

[0111]

[0112] Among them, the activity unit of UNG enzyme is 1U / μl, the activity unit of Taq enzyme is 5U / μl, and the activity unit of RT enzyme is 200U / μl.

[0113] The packaging and content of each component of the kit are shown in Table 8:

[0114] Table 8:

[0115] product name Tube category Mark volume ( / tube) Actual dispensing volume ( / tube) BCR-ABL PCR reaction solution 1.5ml purple cap screw tube 160μl 170μl BCR-ABL PCR reference reaction 1.5ml orange cap screw tube 160μl 170μl Internal reference PCR reaction solution 1.5ml gray cap screw tube 160μl 170μl Internal reference reaction solution 1.5ml brown cap screw tube 160μl 170μl mixed enzyme solution 1.5ml yellow cap screw tube 80μl 90μl control enzyme solution 1.5ml white cap screw tube 8...

Embodiment 2

[0116] Embodiment 2: the accuracy detection of the kit of this application

[0117] Adopt the test kit of embodiment 1 to detect the accuracy reference product:

[0118] Wherein, the preparation method of the accuracy reference product is as shown in Table 9:

[0119] Table 9

[0120]

[0121]

[0122] The accuracy reference product that is prepared is repeated experiment 3 times, and the experimental result obtained is as shown in table 10:

[0123] Table 10

[0124]

[0125] The detection kit of the present application includes accurate quality control samples containing BCR-ABL-210 positive peripheral blood cell lysates and plasmids, and the positive coincidence rate is 100%.

Embodiment 3

[0126] Example 3: Analysis-specific detection of the kit of the present application

[0127] Adopt the kit of embodiment 1 to detect specific reference substance:

[0128] Wherein, the preparation method of specific reference product is shown in Table 11:

[0129] Table 11

[0130] Numbering Preparation method N1 BCR-ABL negative peripheral blood cell lysate, diluted to 5 × 10 with Trizol 4 cells / ml

N2 HL60 cell line lysate, diluted to 5×10 with Trizol 4 cells / ml

N3 Lysate of Jurket cell line, diluted to 5×10 with Trizol 4 cells / ml

[0131] The prepared analysis-specific reference product was repeated for 3 times, and the experimental results obtained are shown in Table 12:

[0132] Table 12

[0133]

[0134]

[0135] The detection kit of the present application includes BCR-ABL-210 negative peripheral blood cell lysate, BCR-ABL-210 negative cell line lysate specific working reference sample, and its negative coincidence r...

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Abstract

The invention relates to the field of fusion gene detection, and in particular, relates to a kit for detecting a human BCR-ABL fusion gene. The kit adopts one-step reverse transcription to detect thehuman BCR-ABL fusion gene. The kit includes a nucleic acid amplification reagent, wherein the nucleic acid amplification reagent includes a BCR-ABL PCR reaction solution, an internal reference PCR reaction solution and a mixed enzyme solution. The kit combines the fluorescent PCR with the reverse transcription technology, and adopts a one-step method to detect the expression level of the BCR-ABL210 fusion gene in a sample, and has the technical advantages of high specificity, high accuracy and high sensitivity.

Description

technical field [0001] This application relates to the field of detection of fusion genes, in particular, to a kit for detecting human BCR-ABL fusion genes and a method for using the same. Background technique [0002] Leukemia is a malignant tumor of the hematopoietic system, which is characterized by the malignant proliferation of one or more blood cells in the hematopoietic system such as bone marrow and lymph nodes, and infiltrates various tissues and organs in the body, resulting in the inhibition of normal hematopoietic tissue cells and the production of various diseases. symptom. Clinically, leukemia is often characterized by fever, hemorrhage, anemia, and enlarged liver, spleen, and lymph nodes. Leukemia can generally be divided into acute leukemia (AL) and chronic leukemia (CL) according to the natural course of the disease and the degree of cell development. According to the morphology and biochemical characteristics of white blood cells, it can be divided into ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/166C12Q2600/118
Inventor 谢立群陈嘉铮李云航陶慧卿
Owner 苏州云泰生物医药科技有限公司
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