Simplex methylmalonic acidmia gene mutation detection primer pair and kit

Abstract

The invention discloses a simple methylmalonic acidmia gene mutation detection primer pair and a kit, the primer pair comprises three primer pairs aiming at a Mut gene, and the nucleotide sequences of the three primer pairs are respectively shown as SEQ ID NO.1-SEQ ID NO.6. The invention further discloses a kit for detecting the mutation of the simple methylmalonic acidmia gene. The kit comprises a Mut gene detection solution, an amplification reaction solution, a sequencing detection solution, a negative reference substance and a positive reference substance, the Mut gene detection liquid comprises three primer pairs aiming at the Mut gene. According to the detection liquid of the kit, only three groups of primer pairs aiming at three hot spot mutation regions are designed, so that the mutation detection of the simple methylmalonemia gene can be completed, the mutation sites of the three hot spot mutation regions of the gene can be quickly, accurately and sensitively detected, and the experimental result is good in repeatability and high in precision.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer pair and a kit for detecting simple methylmalonic acidemia gene mutations. Background technique [0002] Methylmalonic acidemia (MMA) is a kind of hereditary metabolic disease caused by methylmalonyl-CoA mutase (methylmalonyl-CoA mutase, MCM) deficiency or cobalamin metabolism disorder. The massive accumulation of metabolites such as methylmalonic acid and methyl citrate (MCA) leads to metabolic disorders and multiple organ damage. According to whether it is combined with homocysteinemia, MMA is divided into combined type and simple type, both of which are inherited in an autosomal recessive manner. [0003] Combined MMA belongs to cobalamin metabolism disorders, including multiple subtypes. The cobalamin C (cb1C) type caused by the mutation of the cobalamin metabolism-related C gene (MMACHC) is The most common, accounting for more than 80%. Simple MMA is...

AI Technical Summary

Problems solved by technology

[0004] In recent years, the level of tandem mass spectrometry, enzyme detection and gene detection has developed rapidly, and relying solely on GC / MS technology and plasma amino acid detection can no longer meet the needs of disease typing

Gene mutation analysis is the most reliable basis for identification and typing. Because the mut type does not respond to vitamins, the prognosis is worse than that of the cblC type. Therefore, the identification of the mut type and the cblC type is very important clinically. However, at present, there is no specific MMA gene mutation. For the detection scheme, there is only next-generation sequencing for the detection of gene mutations in MMA patients. This method has a large throughput and high coverage, but it is expensive and technically demanding, and it is difficult to promote clinically.

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