Detection kit for fragile X syndrome FMR1 gene

Abstract

The invention discloses a fragile X syndrome FMR1 gene detection kit. The fragile X syndrome FMR1 gene detection kit comprises a specific PCR (Polymerase Chain Reaction) amplification primer for amplifying a CGG repeating region of an FMR1 gene, a third primer for randomly anchoring the CGG repeating region, a DNA (Deoxyribose Nucleic Acid) polymerase with high thermal stability, a control standard substance, a high-GC reaction buffer solution, dNTPs (Deoxyribose Nucleic Acid) and Taq Extender Addive. Various sequences with different sizes are amplified through a three-primer PCR (Polymerase Chain Reaction) reaction, an amplification product is subjected to capillary electrophoresis, the CGG repetition number is calculated through comparison with a standard substance, and female gene heterozygosis and homozygosis can be accurately judged. The kit is used for group screening and prenatal diagnosis of the fragile X syndrome, normal and pre-mutation carriers, full mutation patients, female patients and carriers can be accurately detected, and the kit has the characteristics of rapidness, high accuracy, high flux, economy and safety. Therefore, the kit has a good clinical application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a detection kit for fragile X syndrome FMR1 gene. Background technique [0002] Fragile X syndrome is an X-linked recessive genetic disease mainly characterized by mental retardation. Among the genetic diseases related to mental retardation, its incidence rate is second only to trisomy 21, and it is the most common known disease. X-linked genetic disease is also the single gene genetic disease with the highest incidence rate in humans. Fragile X syndrome is closely related to the abnormal expansion of the n-repeat sequence (CGG) at the 5' end of the FMR1 gene at Xq27.3. There is a trinucleotide repeat sequence mainly composed of CGG (cytosine-guanine-guanine) in the non-coding region near the 5' end of exon 1 of FMR1 gene. The (CGG)n repeat sequence copy number shows great heterogeneity in the population. The (CGG) n of this sequence in normal people is 6-45; 45-55 CGG is called t...

AI Technical Summary

Problems solved by technology

Existing FMR1 gene detection methods include methylation-specific PCR, PCR amplification detection method, Southern blot method, etc., but the above methods cannot detect pre-mutation carriers, and it is difficult to amplify pre-mutation carriers or full-mutation patients. Product, cumbersome operation, and low throughput limit its clinical application; three-primer PCR has a super-strong amplification system, which can avoid the difficulty of conventional PCR amplification due to the high GC content of the sequence. With the increase of (CGG)n repeat number increase, the problem that the amplification ability of PCR decreases

Usually, it is difficult for pre-mutation carriers or patients with full mutations to amplify PCR products, and women have two X chromosomes, which increases the difficulty of PCR detection

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