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Reagent kit for detecting human WT1 fusion genes, and application method of reagent kit

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 unsatisfactory FCM sensitivity and achieve the effects of improved specificity, stable concentration, and reduced pollution.

Pending Publication Date: 2019-12-10
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

Therefore, the sensitivity of FCM is not ideal

Method used

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  • Reagent kit for detecting human WT1 fusion genes, and application method of reagent kit
  • Reagent kit for detecting human WT1 fusion genes, and application method of reagent kit
  • Reagent kit for detecting human WT1 fusion genes, and application method of reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] A test kit for detecting human WT1 fusion gene, the composition of which is shown in Table 5:

[0096] table 5:

[0097]

[0098]

[0099] 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.

[0100] The packaging and content of each component of the kit are shown in Table 6:

[0101] Table 6:

[0102]

Embodiment 2

[0103] Example 2: Linear Detection

[0104] Select 100 μl each of the WT1 plasmid with a determined concentration and the plasmid of the internal reference gene to make a high-concentration DNA sample (L0). The L0 sample was serially diluted with a gradient of 1:10 to obtain two sets of linear quality control substances L1, L2, L3 and L4, respectively. The specific composition is shown in Table 9 and Table 10.

[0105] The logarithmic value of the concentration is Y, and the mean value of CT is X, and linear fitting is carried out to calculate the linear correlation coefficient r. The specific experimental results are shown in Table 7 and Table 8:

[0106] Table 7

[0107] WT1 Linear Quality Control Enter the number of reaction templates (copies) WT1-L0 5×10 6

[0108] Table 8

[0109] Internal reference gene linear quality control Enter the number of reaction templates (copies) Internal reference-L0 5×10 6

[0110] Operate according t...

Embodiment 3

[0111] Example 3: Detection of the accuracy of the kit of the present application

[0112] Adopt the test kit of embodiment 1 to detect the accuracy reference product: wherein, the preparation method of the accuracy reference product is as shown in Table 9:

[0113] Table 9

[0114]

[0115]

[0116] The prepared accuracy reference product is detected, and the experimental results obtained are shown in Table 10:

[0117] Table 10

[0118]

[0119] Using the test kit of the present application to detect the accuracy reference product, the positive coincidence rate is 100%.

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Abstract

The application of the invention relates to the field of detection of fusion genes, in particular to a reagent kit for detecting human WT1 fusion genes. According to the reagent kit disclosed by the application of the invention, reverse transcription is performed by a one-step method, namely that mRNA in a sample is subjected to reverse transcription to form cDNA through a specific primer and an enzyme, the cDNA obtained through reverse transcription directly enters a downstream fluorescent quantitation PCR, and RNA of a WT1 gene in a clinical peripheral blood specimen is subjected to quantitative detection, so that the reagent kit assists myoblastosis clinical diagnosis and guides relevant myoblastosis patients to choose clinical medicines, and has technical advantages of being high in specificity, high in accuracy and lowest in detection quantity.

Description

technical field [0001] This application relates to the field of fusion gene detection, in particular, to a kit for detecting human WT1 fusion gene and its application method. 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 various symptoms. Clinically, leukemia is often characterized by fever, hemorrhage, anemia, and enlarged liver, spleen, and lymph nodes. [0003] Clinical studies in recent years have shown that about 50% of leukemias have certain chromosomal aberrations, such as translocations, deletions, and insertions. Chromosomal translocation aberrations form related fusion genes in most cases. The detection and monitoring of these fusion genes is beneficial to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/166C12Q2600/118C12Q2531/113C12Q2563/107C12Q2521/107
Inventor 熊慧陶慧卿童光铨陈嘉铮徐任俞浩
Owner 苏州云泰生物医药科技有限公司
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