Primer group and kit for single-tube detection of human spinal muscular atrophy

Abstract

The invention discloses a primer group and a kit for detecting copy number and point mutation of a single-tube human spinal muscular atrophy related gene and typing a carrier. The nucleotide sequencesof the primer group are shown as SEQ ID NO. 1 to 38. The primer group is reasonable in design and high in specificity. The amplification bias is reduced, and the quantification accuracy is ensured. An identification sequence is introduced to realize identification of the conversion condition of the seventh exon of the SMN1 and the SMN2. UDG enzyme is introduced, and amplification products are cut, so as to avoid pollution. Clinical applications such as SMN1 / 2 gene copy number detection of human amyotrophy, SNP site detection of SMN1 / 2 Chinese population, NAIP and GTF2H2 gene copy number detection, typing of patients (I- VI types) and carriers ('1 + 0' and '2 + 0') and the like are simultaneously realized by the single tube. The method and the kit are rapid in detection, accurate in result, appropriate in cost, high in clinical occupancy of applicable instruments and suitable for clinical application.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis, in particular to a primer set and a kit for single-tube detection of human spinal muscular atrophy. Background technique [0002] Spinal muscular atrophy (SMA) is a recessive genetic disease characterized by progressive degeneration and loss of spinal cord anterior horn cells (ie, lower motor neurons) and brain stem cell nuclei, resulting in muscle weakness and atrophy. According to multi-ethnic studies on SMA, the overall carrier frequency of SMA is 1 / 40-1 / 100, the incidence rate is about 1 / 11000, and the carrier rate of the Chinese population is about 1 / 50. [0003] Survival motor neuron genes (SMN, NCBI #6606) are located at the 5q13 position of human chromosome 5, including survival motor neuron 1 (SMN1) on the telomeric side and survival motor neuron 2 (SMN2) on the centromere side . SMN1 and SMN2 are highly homologous, both contain 9 exons, and there are only 5 base differences. [0004...

AI Technical Summary

Problems solved by technology

[0014] The purpose of the present invention is to overcome the shortcomings of the prior art that the current human motor neuron gene detection technology and products cannot simultaneously meet the multiple clinical application requirements of patient diagnosis, phenotype typing, and carrier screening, and provide a simple-to-operate, Low-cost, fast and reliable primers and reagents for the single-tube detection based on the combination of QF-PCR technology and CE technology to simultaneously realize the copy number of human muscular dystrophy-related genes and the detection of SNP sites of SMN1 / 2 gene Chinese population, and carrier typing The box can simultaneously detect the copy number of exon 7 of SMN1 and SMN2 genes (including conversion), the deletion of exon 4 and exon 5 of NAIP gene, and the deletion of exon 10 of GTF2H2 gene in a single tube. Exon number deletion, SMN1 / 2 gene SNP loci in Chinese population, "2+0" carrier hint loci and STR loci analyzed by their parents, etc.

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