A detection method and a detection kit for PKD1 gene and PKD2 gene mutation

Abstract

The invention discloses a detection method and a detection kit for PKD1 gene and PKD2 gene mutation. Amplification is performed by creatively designing multiple pairs of specific long fragment PCR primers, and when amplification is performed by adopting the human genome DNA as a template, amplification of pseudogenes with high homology can be avoided, and therefore the appearance of the false positive and false negative result brought by the pseudogenes can be avoided, and the detection accuracy of mutation of the PKD1 gene and PKD2 gene can be greatly improved. A high-throughput sequencing technique is combined, and the method and the kit when compared with a traditional medical exon trapping technique, can simplify a technical process, reduce the detection cost and shorten the detectionperiod.

Description

technical field [0001] The invention relates to the field of biochemical detection, in particular to a detection method and a detection kit for PKD1 gene and PKD2 gene mutations. Background technique [0002] The PKD1 gene is located on the short arm of human chromosome 16 (16p13.3-p13.12). The gene length is 52kb and contains 46 exons. Its structure is as follows: figure 2 As shown, the length of its mRNA is 14kb, and the encoded protein is called polycystin-1 (polycystin-1). The PKD1 gene has 6 homologous pseudogenes on chromosome 16. The homology between the pseudogene and exon 1-33 of the PKD1 gene is as high as 97.7%, and about 80% of the pathogenic mutations occur in this region. Moreover, the GC content of some DNA sequences is as high as 85%, and gene amplification is very difficult. There are many types of gene mutations, involving the entire gene, and there are no mutation hotspots. Therefore, it is difficult to specifically detect PKD1 gene mutations. PKD2 is lo...

AI Technical Summary

Problems solved by technology

However, traditional medical explicit capture technology and other gene detection technologies are prone to missed detection, and it is very easy to amplify pseudogenes with high homology when amplifying PKD1 and PKD2 genes, so there will be false positive and false negative results

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