Why the study?
CYP2C19 loss of function impairs clopidogrel activation and increases platelet aggregation, but the most efficient technique for identifying the CYP2C19*2 polymorphism remained to be determined.
Is RFLP-PCR more efficient than ARMS-PCR for identifying the CYP2C19*2 polymorphism in patients with atherosclerosis?
Is RFLP-PCR more efficient than ARMS-PCR for identifying the CYP2C19*2 polymorphism in patients with atherosclerosis?
RFLP-PCR is significantly more effective than ARMS-PCR for detecting the CYP2C19*2 polymorphism, a key genetic variant affecting clopidogrel metabolism.
Discordant CYP2C19*2 detection by RFLP-PCR versus ARMS-PCR warrants genotyping caution; leaves open validated method for clopidogrel pharmacogenomics.
The CYP2C19*2 polymorphism is the result of a point mutation leading to an alternative splicing defect, generating an enzyme with a reduced function. The enzyme CYP2C19 is responsible for the activation of the prodrug Clopidogrel and the loss of its function leads to an increase in platelet aggregation. We compared the ARMS-PCR (Amplification Refraction Mutation System) and RFLP-PCR (Restriction Fragment Length Polymorphism) to determine which of the techniques was more efficient for identifying the CYP2C19*2 polymorphism. We analyzed 200 samples from patients with atherosclerosis and 100 samples from a control group, free from the disease. The polymorphic allele was not observed in any of the samples analyzed by the ARMS-PCR technique, but was present in more than 70% of the samples when analyzed by RFLP-PCR. Thus, RFLP-PCR generated more satisfactory results for the analysis of CYP2C19*2 polymorphism, and it should be used, despite the lower cost of ARMS-PCR, as the latter did not detect this polymorphism.
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Costa et al. (2019) studied this question.
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