Fluorescent quantitative PCR (Polymerase Chain Reaction) kit for diagnosing human spinal muscular atrophy

Abstract

The invention relates to a fluorescent quantitative PCR (Polymerase Chain Reaction) kit for diagnosing human spinal muscular atrophy. The fluorescent quantitative PCR kit comprises amplification primers and fluorescent probes. The fluorescent quantitative PCR kit is characterized in that the amplification primers include a pair of shared primers used for specifically amplifying SMN1 (Survival Motor Neuron 1) and SMN2 (Survival Motor Neuron 2) genes, a pair of primers used for specifically amplifying an NAIP (Neuronal Apoptosis Inhibitory Protein) gene and a pair of primers used for specifically amplifying a reference sequence; the fluorescent probes include a fluorescent probe used for specifically detecting the SMN1 gene, a fluorescent probe used for specifically detecting the SMN2 gene, a fluorescent probe used for specifically detecting the NAIP gene and a fluorescent probe used for specifically detecting the reference sequence; the reference sequence is as shown in SEQ NO.14,and is positioned in a GRCh37 / hg19chr5:71107506-71107618 interval positioned on the 78kb position of the telomere side of the NAIP gene. The fluorescent quantitative PCR kit disclosed by the invention remarkably enhances the accuracy and stability of diagnosis by taking the DNA (Desoxvribose Nucleic Acid) sequence positioned at the telomere side of the NAIP gene as the reference sequence.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to reagents for nucleic acid determination or detection methods. Background technique [0002] Spinal muscular atrophy (SMA) is a group of common autosomal recessive genetic diseases, ranking second among fatal autosomal recessive genetic diseases, and the incidence rate of live births is 1 / 6000~ 1 / 10000. Clinically, SMA is generally divided into 4 types: Type Ⅰ (also known as Werding-Hoffman disease) is the most severe subtype (severe), accounting for about half of SMA patients, with acute onset and rapid progression, usually within 6 months of birth Onset within 2 years; SMA type Ⅱ is chronic infantile type (intermediate type), usually onset within 7 to 18 months, and most of them can survive 10 to 20 years old; SMA type Ⅲ (also known as Wohlfart-Kugelberg- Welander disease) is juvenile type (mild type), and the onset occurs after 18 months of birth. The disease develops slowly and c...

AI Technical Summary

Problems solved by technology

At present, the routine detection of SMA in clinical practice mainly adopts the Multiple Ligation-dependent Probe Amplification (MLPA) detection kit from MRC Company in the Netherlands. ), requires specific analytical instruments, takes a long time (about 24 hours), and has a heavy burden on patients, making it difficult to carry out large-scale population screening and technology promotion in clinical practice

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