High-resolution melting analysis to screen the ST18 gene functional risk variant for pemphigus vulgaris: The occasion to open a debate on its usefulness in clinical setting

Maria De Bonis; Elisa De Paolis; Giovanni Luca Scaglione; Luca Fania; Maria Michela Lavieri; Cinzia Mazzanti; Giovanni Di Zenzo; Angelo Minucci; Ettore Capoluongo

doi:10.1016/j.yexmp.2019.03.009

High-resolution melting analysis to screen the ST18 gene functional risk variant for pemphigus vulgaris: The occasion to open a debate on its usefulness in clinical setting

Exp Mol Pathol. 2019 Jun:108:57-63. doi: 10.1016/j.yexmp.2019.03.009. Epub 2019 Mar 26.

Authors

Maria De Bonis¹, Elisa De Paolis², Giovanni Luca Scaglione³, Luca Fania⁴, Maria Michela Lavieri⁵, Cinzia Mazzanti⁴, Giovanni Di Zenzo⁶, Angelo Minucci⁷, Ettore Capoluongo⁸

Affiliations

¹ UOC Laboratory Medicine, Genetics and Toxicology, IDI IRCCS, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy; Polo Scienze per Immagini, di Laboratorio e Infettivologiche, "Fondazione Policlinico Agostino Gemelli" - IRCCS, Rome, Italy. Electronic address: [email protected].
² UOC Laboratory Medicine, Genetics and Toxicology, IDI IRCCS, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy; Polo Scienze per Immagini, di Laboratorio e Infettivologiche, "Fondazione Policlinico Agostino Gemelli" - IRCCS, Rome, Italy.
³ UOC Laboratory Medicine, Genetics and Toxicology, IDI IRCCS, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy.
⁴ Department of Dermatology - IDI IRCCS, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy.
⁵ Unit of Dermatology - Ospedale Cristo Re, Via delle Calasanziane, Rome, Italy.
⁶ Molecular and Cell Biology Laboratory IDI-IRCCS, FLMM, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy.
⁷ Polo Scienze per Immagini, di Laboratorio e Infettivologiche, "Fondazione Policlinico Agostino Gemelli" - IRCCS, Rome, Italy.
⁸ UOC Laboratory Medicine, Genetics and Toxicology, IDI IRCCS, Fondazione Luigi Maria Monti, Via Monti di Creta 104, Rome, Italy; Polo Scienze per Immagini, di Laboratorio e Infettivologiche, "Fondazione Policlinico Agostino Gemelli" - IRCCS, Rome, Italy. Electronic address: [email protected].

PMID: 30926339
DOI: 10.1016/j.yexmp.2019.03.009

Abstract

The ST18 -497-65050 T > C polymorphisms (rs17315309) exhibit a very strong association in the pathogenesis of Pemphigus Vulgaris (PV) and could represent a new potential molecular target for the treatment of disease. The present study aimed to establish a low-cost, sensitive and reliable assay using high-resolution melting curve analysis (HRMA) on magnetic induction rotor-based platform, the Magnetic Induction Cycler (MIC) (Bio molecular Systems). HRMA assay was able to identify easily and unambiguously the c.-497-65050 T > C genotypes evaluating melting curve shape and melting temperature (T_m). The results of HRMA were validated by direct DNA sequencing. The HRMA is rapid, sensitive, low-cost and high-throughput assay to screen the rs17315309 variant and could be used in clinical diagnostic laboratories.

Keywords: High Resolution melting analysis; Pemphigus vulgaris; ST18.

MeSH terms

Genetic Predisposition to Disease / genetics*
Genotype
Humans
Molecular Diagnostic Techniques
Nucleic Acid Denaturation*
Pemphigus / diagnosis
Pemphigus / genetics*
Polymorphism, Single Nucleotide*
Repressor Proteins / genetics*
Reproducibility of Results
Risk Factors
Sensitivity and Specificity
Sequence Analysis, DNA / methods

Substances

Repressor Proteins
ST18 protein, human