Design and optimization of 18-gene Ion AmpliSeq panel of Next-generation sequencing for gene mutation analysis causing pain insensitivity

Abstract

Pain insensitivity is a unique phenotypic condition inherited in an autosomal dominant or recessive Mendelian form. Affected individuals suffer the loss of feeling to pain after exposure to high or low temperature. Recurrent mouth ulcers, face flushing, recurrent periods of elevated body temperature, and unexplained sweating are signs of a sensory neuropathy of mild autonomic activity in kids. Pain sensitivity deficiency is characterized by a loss of deep pain and temperature sensation, but natural reasoning skills and most other neural responses remain unchanged. Unfortunately, the spectrum of mutations causing pain insensitivity in Arabs, which is the prerequisite for prenatal and pre-implantation prevention regimes, is not yet determined. Therefore, this study's goal was to develop a high-throughput mutation screening method for gene mutation analysis causing pain insensitivity using the Ion Torrent Next-generation personal genome analyzer based on semiconductor nano-chips technology. We've created a panel that includes 18 genes linked to pain insensitivity. The panel was created using Ion Torrent's Ion AmpliSeq next-generation sequencing technology (NGS). Furthermore, two healthy human control samples were used to optimize all NGS procedures successfully. The molecular method should offer a fast, easy, and cost-effective, reliable mutation scanning/screening method that may also be applied in the future for prenatal and pre-implantation genetic diagnosis to offer families at risk of having a child with pain insensitivity opportunity to give birth to an unaffected child.

Keywords

Pain insensitivity, mutation, next-generation sequencing, peripheral neuropathy.

How to Cite