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Detection kit and ARMS (amplification refractory mutation system)-QPCR (quantitative polymerase chain reaction) detection method for beta-mediterranean anemia beta17 (A-T) point mutation

A technology of thalassemia and detection kits, applied in the field of molecular biology, can solve the problems of unsuitable detection of large sample size, low detection sensitivity, long detection time, etc., and achieve small error, high detection sensitivity and high accuracy Effect

Active Publication Date: 2014-02-19
珠海辉睿生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technical effect describes a new way to test beta thalsemias by measuring changes caused by certain genes that are involved during their development or disease process called Thymosin-1α(Thy1). By doing this quickly and accurately, researchers may better identify these diseases early on before they develop symptoms like seizures.

Problems solved by technology

In beta thalloid syndrome, patients who suffer from type 2 dihydroxyuraccharidosis exhibit symptoms such as seizures due to reduced levels of coenzyme Q10 activity associated with decreased production of neurotransmitters called cytosine monophosphate guanylate cyclotransferases (CNQ1 - CYTB). These defects lead to deficiency of enzyme activities involved in metabolism processes like gamma glutamisceratitis virus (GAE)-associating polyglucocerebrosides protein disulfursibocytose dysfunction pathway (CDH362), sickling cellular cresy bodies (SCUBB1), autochondria chromatinum pyrophosphorridium complex (AMLPZ)/dithiothreuxene/pyrrolimodynia platinyoxane (DDPT)) systems, including methyllanthanocephalus bacterial cells containing histone variants, and various types therapy agents based on these nucleuses.

Method used

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  • Detection kit and ARMS (amplification refractory mutation system)-QPCR (quantitative polymerase chain reaction) detection method for beta-mediterranean anemia beta17 (A-T) point mutation
  • Detection kit and ARMS (amplification refractory mutation system)-QPCR (quantitative polymerase chain reaction) detection method for beta-mediterranean anemia beta17 (A-T) point mutation
  • Detection kit and ARMS (amplification refractory mutation system)-QPCR (quantitative polymerase chain reaction) detection method for beta-mediterranean anemia beta17 (A-T) point mutation

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Embodiment 1

[0075] Such as Figure 1-7 As shown, a detection kit for β-thalassemia β17 (A→T) point mutation is characterized in that it includes PCR reaction solution I, PCR reaction solution II, mixed enzyme solution, DNA extraction solution, negative control solution and positive controls;

[0076] The PCR reaction solution I includes an upstream primer I, a probe and a downstream primer;

[0077] The PCR reaction solution II includes an upstream primer II, a probe and a downstream primer;

[0078] The mixed enzyme solution includes DNA polymerase, sterilized ultrapure water I and chromogenic solution;

[0079] The DNA extraction solution includes phenol and chloroform;

[0080] The negative control solution is a Tris-EDTA buffer solution that does not contain the β-thalassemia β17 (A→T) gene, the concentration is 0.01M, and the pH is 8.00;

[0081] The positive control substance is a DNA synthesis positive template using β-thalassemia β17 (A→T), including a mutant positive template...

Embodiment 2

[0134] Such as Figure 1-7 As shown, a detection kit for β-thalassemia β17 (A→T) point mutation,

[0135] The negative control solution is a Tris-EDTA buffer solution that does not contain the β-thalassemia β17 (A→T) gene, the concentration is 0.015M, and the pH is 8.02;

[0136] The contents of the sterilized ultrapure water II, buffer, magnesium chloride, dNTPs, upstream primer I, upstream primer II, downstream primer, probe, DNA polymerase, sterilized ultrapure water I and chromogenic solution are as follows:

[0137] The volume of sterilized ultrapure water II is 4.8 μl / tube;

[0138] The concentration of the buffer solution is 15×Buffer, and the volume is 4 μl / tube;

[0139] Magnesium chloride Concentration is 30 mM Volume is 4 μl / tube;

[0140] The concentration of dNTPs is 3.0mM and the volume is 4 μl / tube;

[0141] The concentration of upstream primer Ⅰ is 100 μM and the volume is 0.2 μl / tube;

[0142] The concentration of upstream primer II is 100 μM and the volu...

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Abstract

The invention discloses a detection kit and an ARMS (amplification refractory mutation system)-QPCR (quantitative polymerase chain reaction) detection method for beta-mediterranean anemia beta17 (A-T) point mutation, and is characterized in that: the detection kit includes a PCR (polymerase chain reaction) solution I, a PCR solution II, a mixed enzyme solution, a DNA extraction solution, a negative comparison solution and a positive comparison product; and the ARMS-QPCR detection method includes sample collection, ARMS primer design, sample processing and other steps. By use of the detection kit for the beta-mediterranean anemia beta17 (A-T) point mutation, the detection of a to-be-detected beta-mediterranean anemia beta17 (A-T) point mutation DNA sample can be completed only for 3 hours, at the same time, a real-time fluorescent quantitative PCR method for detection of the beta-mediterranean anemia beta17 (A-T) point mutation is strong in gene specificity, high in detection sensitivity, simple, small in error, and high in accuracy.

Description

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Claims

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Application Information

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Owner 珠海辉睿生物科技有限公司
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