Rapid detection of Mmalton α<inf>1</inf>-antitrypsin deficiency allele by real-time PCR and melting curves in whole blood, serum and dried blood spot samples

Irene Belmonte, Luciana Montoto, Marc Miravitlles, Miriam Barrecheguren, Cristina Esquinas, Esther Rodríguez, Marina Giralt, Francisco Rodríguez-Frías

Research output: Contribution to journalArticleResearchpeer-review

13 Citations (Scopus)

Abstract

© 2016 by De Gruyter 2016. Background: α1-Antitrypsin deficiency (AATD) is an autosomal codominant disorder associated with a high risk of developing lung and liver disease. The most common deficient alleles are known as Z and S. However, another deficient variant, called Mmalton, which causes a deficiency similar to variant Z, is considered to be the second cause of severe AATD in Spain. Nevertheless, the Mmalton allele is not recognizable by usual diagnostic techniques and therefore, its real prevalence is underestimated. We describe a rapid real-time PCR and melting curves assay designed for the detection of Mmalton AATD. Methods: We tested the applicability of this new technique for the identification of the Mmalton allele in AATD screening using whole blood, dried blood spot (DBS) and serum samples. Mmalton heterozygote and homozygote samples and samples without this allele were included in the study. Results: This new assay is able to detect homozygous and heterozygous genotypes in the same reaction and in a single step, giving matching results with those obtained by SERPINA1 gene sequencing. Conclusions: This technology is optimal for working with small amounts of DNA, such as in DBS and even with residual DNA present in serum samples, allowing improvement in routine algorithms of AATD diagnosis or large-scale screening. This method will be useful for obtaining more in depth knowledge of the real incidence of the Mmalton variant.
Original languageEnglish
Pages (from-to)241-248
JournalClinical Chemistry and Laboratory Medicine
Volume54
Issue number2
DOIs
Publication statusPublished - 1 Jan 2016

Keywords

  • dried blood spot
  • FRET probes
  • melting curves
  • Mmalton variant
  • α -antitrypsin deficiency 1

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