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Updated: Jul 17, 2026
Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
Published on: August 8, 2016
Fast CYP2E1 genotyping using automated fluorescent detection
Claudia Innocenti1, A Accorsi, V Cerreta
1Center for Applied Biomedical Research (CRBA), Sant'Orsola-Malpighi Hospital, Bologna, Itlay.
Background:
Among polymorphic genes coding for xenobiotic-metabolizing enzymes, the ethanol-inducible CYP2E1 gene (1667 bp) is known to play a major role in the metabolism ofseveral chemicals.
Objectives:
In order to apply large-scale genotyping, we explored the use of a Single Nucleotide Primer Extension (SNuPE) assay coupled with automated fluorescent detection to assess the presence of low-frequency CYP2E1*5B (c2) allele.
Methods:
a classic PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method specific for polymorphic5'-flanking region of CYP2E1 gene was tested in conjunction with a newly developed accelerated SNuPE assay.
Results:
compared to the classic PCR-RFLP method, the accelerated SNuPE assay proved to be both sensitive and specific for fast CYP2E1 genotyping.
Conclusions:
automated fluorescent methods as SNuPE assay are usefulfor public health perspectives, allowing rapid genotyping of metabolic genes in large population studies in clinical or epidemiological settings.
