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.

Fast CYP2E1 genotyping using automated fluorescent detection

ACCORSI, ANTONIO;MANTOVANI, VILMA;VIOLANTE, FRANCESCO SAVERIO
2006

Abstract

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.
LA MEDICINA DEL LAVORO
Innocenti C.; Accorsi A.; Cerreta V.; Mantovani V.; Violante F.S.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/37880
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