title: Rapid PCR real-time genotyping of M-Malton alpha1-antitrypsin deficiency alleles by molecular beacons.
abstract: Alpha1-Antitrypsin deficiency is an autosomal codominant inherited disorder, with increased risk of developing lung and liver disease. The large majority of subjects affected by alpha1-antitrypsin deficiency carry the PIZZ or PISZ genotypes, which can be easily detected using several molecular methods. Another pathologic allele, the M-Malton variant (also known as Mnichinan and Mcagliari), can mimic the Pi Z clinical phenotype, but this alpha1-antitrypsin deficiency variant is not easily recognizable and, therefore, seems to be more under-recognized than the Z or S alleles. We report the development of a rapid qualitative fluorescent real-time PCR assay designed for the detection of the M-Malton alpha1-antitrypsin deficiency alleles using 2 specific molecular beacons. The assay is able to detect in a single tube the homozygous as well the heterozygous genotypes. The procedure combines the great sensitivity of the polymerase chain reaction, the specificity provided by allele-specific molecular beacons, and the throughput of a multi-color fluorescence detection procedure. This technique will be useful for research and molecular diagnostic laboratories involved in the study of alpha1-antitrypsin deficiency-related diseases.
authors.affiliation: Dipartimento di Chirurgia e Scienze Odontostomatologiche Universita' degli Studi di Cagliari, Cagliari Italy.
authors.forename: Germano
authors.initials: G
authors.lastname: Orrù
authors.forename: Gavino
authors.initials: G
authors.lastname: Faa
authors.forename: Sara
authors.initials: S
authors.lastname: Pillai
authors.forename: Luca
authors.initials: L
authors.lastname: Pilloni
authors.forename: Caterina
authors.initials: C
authors.lastname: Montaldo
authors.forename: Gesuina
authors.initials: G
authors.lastname: Pusceddu
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Piras
authors.forename: Pierpaolo
authors.initials: P
authors.lastname: Coni
mesh_terms: D000293:Adolescent; D000328:Adult; D000483:Alleles; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D005260:Female; D005787:Gene Frequency; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D000515:alpha 1-Antitrypsin / Q000235:genetics*; D019896:alpha 1-Antitrypsin Deficiency / Q000235:genetics*
chemical_list: D000515:alpha 1-Antitrypsin