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Malaysian Journal of Medical Sciences
School of Medical Sciences, Universiti Sains Malaysia
ISSN: 1394-195X
Vol. 25, No. 4, 2018, pp. 72-81
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Bioline Code: mj18048
Full paper language: English
Document type: Research Article
Document available free of charge
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Malaysian Journal of Medical Sciences, Vol. 25, No. 4, 2018, pp. 72-81
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A Novel Multiplex PCR-RFLP Method for Simultaneous Genotyping of CYP3A4*4 A>G, CYP3A4*18B G>A and CYP3A4*22 C>T
Abubakar, Murtala Bello; Tan, Huay Lin & Gan, Siew Hua
Abstract
Background: Cytochrome P450 3A enzymes exhibit a variety of physiological roles
and have been reported to be the most predominant enzymes involved in drugs metabolism.
Single nucleotide polymorphisms (SNPs) in the genes that code for these enzymes may result in
functional changes that affect enzyme activity. CYP3A4 is an important enzyme in the metabolism
of many important drugs used in the treatment of breast cancer.
Methods: A total of 94 post-menopausal breast cancer patients were recruited for
the study and their DNA was isolated for polymerase chain reaction (PCR). The primers were
designed using Primer3 software with primer specificities checked via the Basic Local Alignment
Tool (BLAST) database. The primer specificity, functionality and annealing temperature were
first investigated using uniplex PCR protocols, followed by a single multiplex polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The digested
amplification fragments were analysed by gel electrophoresis and subsequently validated by
sequencing.
Results: A multiplex PCR-RFLP method was successfully developed for simultaneous
detection of CYP3A4*4, CYP3A4*18B and CYP3A4*22 in a population of post-menopausal breast
cancer patients.
Conclusion: The technique is simple, cost-effective, time-saving and can be routinely
applied in the identification of SNPs and determination of allelic and genotypic frequencies of
CYP3A4*4, CYP3A4*18B and CYP3A4*22.
Keywords
multiplex PCR-RFLP; single nucleotide polymorphisms; CYP3A4*4; CYP3A4*18B; CYP3A4*22
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© Copyright 2018 - Penerbit Universiti Sains Malaysia Alternative site location: http://www.medic.usm.my/publication/mjms/
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