Real-Time PCR to Detect α-1 Antitrypsin S and Z Alleles in Formalin-Fixed Paraffin-Embedded Tissue

α-1 Antitrypsin (A1AT) deficiency is an autosomal recessive genetic disease with incomplete penetrance that can cause pulmonary and liver disease. Multiple methods are available to determine A1AT genotype using peripheral blood specimens, but none are validated to detect A1AT alleles in formalin-fix...

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Bibliographic Details
Published in:The journal of applied laboratory medicine 2018-07, Vol.3 (1), p.18-25
Main Authors: Pac, Lincoln J, Cheeney, Gregory, Westerhoff, Maria, Konnick, Eric Q, Greene, Dina N, Lockwood, Christina M
Format: Article
Language:English
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Summary:α-1 Antitrypsin (A1AT) deficiency is an autosomal recessive genetic disease with incomplete penetrance that can cause pulmonary and liver disease. Multiple methods are available to determine A1AT genotype using peripheral blood specimens, but none are validated to detect A1AT alleles in formalin-fixed paraffin-embedded (FFPE) tissue. A real-time PCR assay was validated to detect the SERPINA1 S and Z alleles (NM_000295.4: c.863A>T, p.E288V and c.1096G>A, p.E366K, respectively) in FFPE liver tissue using allele-specific dual hybridization probes and melting curve analysis. Validation experiments were performed on genomic DNA samples (n = 11) with A1AT genotypes previously determined by orthogonal methods. The S and Z allele assays accurately genotyped all FFPE validation specimens that had a threshold cycle
ISSN:2576-9456
2475-7241
2475-7241
DOI:10.1373/jalm.2017.025858