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

• Published in: Experimental dermatology 2018-11, Vol.27 (11), p.1304-1308
• Main Authors: Hwang, Joonsung, Lee, Sangku, Kim, Daehwan, Han, Goeun, Soung, Nak Kyun, Cha‐Molstad, Hyunjoo, Lee, Kyung Ho, Ryoo, In Ja, Ahn, Mi Ja, Kim, Sung Tae, Lee, Min Jae, Yoo, Young Dong, Lee, Hee Gu, Hong, Jin Tae, Kim, Hyunjung, Choi, Eung Ho, Kim, Soo‐Chan, Kwon, Yong Tae, Ahn, Jong Seog, Kim, Bo Yeon
• Format: Article
• Language: English
• Subjects: Atopic dermatitis; Dermatitis; DNA probes; Filaggrin; Filaggrin mutation; Fluorescent indicators; Heterozygotes; high‐throughput screening; medical diagnosis; Melting curve; Mutation; Patients; Peptide Nucleic Acid (PNA); Peptide nucleic acids; Skin diseases
• ISSN: 0906-6705; 1600-0625
• DOI: 10.1111/exd.13765

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.