Rapid detection kit for fragile X syndrome

A detection kit and rapid technology, which can be used in the determination/examination of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of high cost, long time, and reduce the incidence of Fragile X syndrome.

Inactive Publication Date: 2017-06-13
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to reduce the incidence of Fragile X Syndrome, based on the understanding of the DNA sequence of the fragile site, methods such as RFLP linkage analysis, DNA hybridiza

Method used

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  • Rapid detection kit for fragile X syndrome
  • Rapid detection kit for fragile X syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: A kind of preparation method of Fragile X Syndrome rapid detection kit

[0016] 1. Primers: Design primers at the CGG repeat of the FRM-1 gene, including upstream primer F and downstream primer R.

[0017] 2. 10×PCR buffer: its components mainly include 50mM Tris-AC (pH9.2), 50mM KAC, 20mM (NH 4 ) 2 SO 4 , 10mM MgCl 2 .

Embodiment 2

[0018] Example 2: The QIAamp DNA Blood Mini Kit (50)-Cat. No. 51104 kit from QIAGEN was used for nucleic acid extraction to obtain the sample to be tested.

Embodiment 3

[0019] Embodiment 3: detection sample

[0020] Tris-AC (pH9.2) 50mM KAC 50mM (NH 4 ) 2 SO 4

20mM MgAC 2

10mM dNTPs 300μM high fidelity enzyme 4U DMSO 6% Betaine 5M F 10pmol R 10pmol dna 50ng

[0021] Its PCR amplification conditions:

[0022]

[0023] After the reaction, the PCR product was subjected to agarose electrophoresis. It can be seen that normal people have clear bands at figure 1 ), while patients with fragile X syndrome had a clear band at >700bp ( figure 1 ). This result shows that this kit can quickly detect Fragile X Syndrome.

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Abstract

The invention discloses a rapid detection kit for a fragile X syndrome. The rapid detection kit for the fragile X syndrome has the technical scheme of (1) designing an upstream primer and a downstream primer in a 5' untranslated region (a CGG repeat region) of an FMR-1 (Fragile X Mental Retardation-1) gene; (2) mixing the upstream primer, the downstream primer with high-fidelity enzyme, 10*PCR (Polymerase Chain Reaction) buffer and the like, and carrying out PCR amplification; (3) carrying out agarose electrophoresis on an amplification product, and judging the approximate copy number of CGG through judging the magnitude of fragments of the amplification product, thus judging whether a detector is a patient suffering from the fragile X syndrome or a carrier carrying the fragile X syndrome or not, wherein the fragments of the amplification product of a normal person are less than 300bp, and if the amplification product of which the fragments are greater than 700bp exists, the person is a pre-mutator or a complete mutator. In a detection method of the invention, the design difficulty of used primers is low, and the synthesis cost of the primers is reduced to a large extent; through a judgment result of an agarose electrophoresis method, the detection cost is further reduced, and the clinical practicability is greatly increased.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid detection kit for fragile X syndrome. Background technique [0002] Fragile X syndrome leads to mental retardation in patients (Martin-Bell syndrome), which is an X-linked recessive genetic disease whose clinical manifestations are mainly mental retardation. Translated region (CGG) n trinucleotide repeat abnormal transmission caused by. The (CGG) repeat copy number of normal people is about 30 copies (90bp), while the repeat copy number of female carriers and normal male passers is increased to 150~500bp, which is called pre-mutation, and the adjacent CpG island is not covered by A Kylation, this premutation (premutation) has no or only mild symptoms. The transformation from pre-mutation to full mutation only occurs during the transmission from mother to offspring: the CGG region of female carriers is unstable, which will lead to the increase of female carriers and male pat...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6883C12Q2600/156C12Q2525/151C12Q2565/125
Inventor 刘淑园陈华云肖湘文丁渭张天海刘孝礼邓利琼陆同山
Owner GUANGZHOU HEAS BIOTECH CO LTD
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