Duplex high resolution melting analysis (dHRMA) to detect two hot spot CYP24A1 pathogenic variants (PVs) associated to idiopathic infantile hypercalcemia (IIH).

Autor: De Bonis M; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy. maria.debonis@policlinicogemelli.it., De Paolis E; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy., Onori ME; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy., Mazzuccato G; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy., Gatto A; UOC Pediatria, Dipartimento Scienze della Salute della Donna, del Bambino E Di Sanità Pubblica- Area Salute del Bambino, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy., Ferrara P; UOC Pediatria, Dipartimento Scienze della Salute della Donna, del Bambino E Di Sanità Pubblica- Area Salute del Bambino, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy., Ferraro PM; UOC Nefrologia, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy.; Università Cattolica del Sacro Cuore, Largo Francesco Vito, 1, 00168, Rome, Italy., Urbani A; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy.; Università Cattolica del Sacro Cuore, Largo Francesco Vito, 1, 00168, Rome, Italy., Minucci A; UOS Diagnostica Molecolare E Genomica, Fondazione Policlinico Universitario A. Gemelli-IRCCS, Largo Agostino Gemelli, 00168, Rome, Italy.
Jazyk: angličtina
Zdroj: Molecular biology reports [Mol Biol Rep] 2021 Apr; Vol. 48 (4), pp. 3303-3311. Date of Electronic Publication: 2021 Apr 17.
DOI: 10.1007/s11033-021-06324-x
Abstrakt: Pathogenic variants (PVs) in CYP24A1 gene are associated with Idiopathic Infantile Hypercalcemia disease (IIH). The identification of CYP24A1 PVs can be a useful tool for the improvement of target therapeutic strategies. Aim of this study is to set up a rapid and inexpensive High Resolution Melting Analysis (HRMA)-based method for the simultaneous genotyping of two hot spot PVs in CYP24A1 gene, involved in IIH. A duplex-HRMA (dHRMA) was designed in order to detect simultaneously CYP24A1 c.428_430delAAG, p.(Glu143del) (rs777676129) and c.1186C > T, p.(Arg396Trp) (rs114368325), in peculiar cases addressed to our Laboratory. dHRMA was able to identify clearly and simultaneously both hot spot CYP24A1 PVs evaluating melting curve shape and melting temperature (T m ). This is the first dHRMA approach to rapidly screen the two most frequent CYP24A1 PVs in peculiar case, providing useful information for diagnosis and patient management in IIH disease.
Databáze: MEDLINE
Externí odkaz: