The Correlation of Two Different Real-Time PCR Devices for the Analysis of CYP2C19 Pharmacogenetic Results

oleh: Alba Alonso Llorente, Josefa Salgado Garrido, Oscar Teijido Hermida, Fabricio González Andrade, Alberto Valiente Martín, Ana Fanlo Villacampa, Jorge Vicente Romero

Format: Article
Diterbitkan: MDPI AG 2023-11-01

Deskripsi

<i>CYP2C19</i> is a highly polymorphic gene responsible for the metabolism of commonly used drugs. <i>CYP2C19*1</i>, the wild-type allele, is associated with normal enzyme activity, whereas <i>CYP2C19*2</i> and <i>CYP2C19*17</i> lead to null and increased enzyme activity, respectively. The use of different instruments to perform the same pharmacogenetic tests should not affect the reliability of the results reported to clinicians, as required by the ISO 15189 standard. Genotyping assays allowed for the identification of gene variants corresponding to the <i>CYP2C19*2</i> and <i>CYP2C19*17</i> haplotypes in 44 selected samples. Each sample was analyzed in duplicate using the Thermo Fisher Taqman Drug Metabolism probes <i>CYP2C19*2</i>: c_25986767_70 (rs4244285) and <i>CYP2C19*17</i>: c_469857_10 (rs12248560). The experiments were performed on two widely used types of real-time PCR analyzers: ABI PRSIM™7500 and QuantStudioTM12KFlex (both from Applied Biosystems, Thermofisher). The data were analyzed in a Thermo Fisher Cloud facility. The analysis was performed independently by two qualified professionals. Both different instruments and analysts’ interpretations were consistent in identifying the native homozygous, heterozygous, and mutant homozygous variants for <i>CYP2C19*2</i> and <i>CYP2C19*17</i>. The results provided by both the primary and backup analyzers showed a perfect correlation. This would allow for the use of the backup analyzer in case the main one is not available.