Peptide nucleic acid (PNA) probe‐based analysis to detect filaggrin mutations in atopic dermatitis patients

Abstract

Atopic dermatitis (AD) is a chronic inflammatory skin disease whose prevalence is increasing worldwide. Filaggrin (FLG) is essential for the development of the skin barrier, and its genetic mutations are major predisposing factors for AD. In this study, we developed a convenient and practical method to detect FLG mutations in AD patients using peptide nucleic acid (PNA) probes labelled with fluorescent markers for rapid analysis. Fluorescence melting curve analysis (FMCA) precisely identified FLG mutations based on the distinct difference in the melting temperatures of the wild-type and mutant allele. Moreover, PNA probe-based FMCA easily and accurately verified patient samples with both heterozygote and homozygote FLG mutations, providing a high-throughput method to reliable screen AD patients. Our method provides a convenient, rapid and accurate diagnostic tool to identify potential AD patients allowing for early preventive treatment, leading to lower incidence rates of AD, and reducing total healthcare expenses.

The role of filaggrin in atopic dermatitis and allergic disease

“…Other forms of measuring filaggrin expression also exist. Hwang et al 46 used PNA (peptide nucleic acid) probes to accurately and rapidly detect FLG mutations. The PNAs are artificial DNA analogues that pair with their complementary DNA targets.…”

“…Thus, through this PNA probe-based fluorescence melting curve analysis they were able to detect both heterozygous and homozygous FLG mutations with precision. 46…”

Potential applications of peptide nucleic acid in biomedical domain

“…PNA is a synthetic molecule that shows highly specific binding to target gene sequence due to which they are an appropriate candidate of interest in biomedical and biotechnological context 3 . Tagalakis et al 120 developed a nanovesicle (∼90 to 140 nm) complex made from giant unilamellar vesicle to efficiently deliver the drug to the target site of gene therapy by therapeutic nucleic acids, as targeted delivery is one of the challenging aspects in PNA‐based gene therapy.…”

Distinct clinical features and serum cytokine pattern of elderly atopic dermatitis in China

“…AD patients from Japan and Korea show a mixed Th2 and Th17 profile of inflammation infiltration in lesional skin, indicating a unique feature of Asian population 10 . Asian AD patients also show more pronounced lichenification in clinical manifestation, apparent epidermal hyperplasia in histology and distinct mutations in filaggrin comparing to white population 11 . However, the clinical features and underlining endotypes of Chinese AD patients across different age groups have not been reported.…”