A Single Multiplexed Allele-Specific Polymerase Chain Reaction for Simultaneous Detection of α₁-Antitrypsin S and Z Mutations

Abstract

Alpha1-antitrypsin (AAT) deficiency is an inherited disorder that can cause lung disease in adults and liver disease in adults and children. The S and Z mutations are the two most common mutations found in the AAT-deficient patients. We have developed a simple multiplexed allele-specific-PCR to detect both the S and Z mutations and the corresponding wild-type alleles. Polymerase chain reaction (PCR) product could be resolved on an agarose gel or using any fluorescent gel detection system. We obtained accurate genotyping results for the four alleles; the S, Z, and their corresponding wild-type alleles for all investigated samples simultaneously. The approach described in this paper is rapid, cost effective, and reliable and can also be adaptable into any laboratory setting because of its simplicity.

Diagnosis of alpha-1 antitrypsin deficiency: Modalities, indications and diagnosis strategy

“…The allelespecific PCR is also commonly used, possibly in multiplex [26]; amplification and hybridization kits with PI S and PI Z allele-specific labelled probes are also marketed [27]. Realtime PCR approaches, also using M, S and Z allele-specific fluorescent probes, have been developed more recently [28,29].…”

“…La PCR allèle-spécifique est également couramment utilisée, éventuellement en multiplex [26] ; des kits d'amplification puis hybridation avec des sondes marquées spécifiques des allèles PI S et PI Z sont aussi commercialisés [27]. Des approches par PCR en temps réel, utilisant également des sondes spécifiques des allèles M, S et Z marquées à la fluorescence, ont été développées plus récemment [28,29].…”

Development and Characterization of Reference Materials for MTHFR, SERPINA1, RET, BRCA1, and BRCA2 Genetic Testing

“…16 The normal allele, M, produces normal levels of the AAT protein. Two mutations, S and Z (representing the Glu264Val and Glu342Lys mutations, respectively), produce low or very low levels of circulating AAT protein, respectively, and patients carrying two of these alleles (ZZ or SZ) are at risk for developing disease related to AATD.…”