BacKgroUND: thiopurine-related hematoxicity is clinically relevant and therapy-limiting in treatment of patients with acute lymphoblastic leukemia (ALL) or inflammatory bowel disease (IBD). For clinical implementation, the Clinical Pharmacogenetics Implementation Consortium (cPic) published peer-reviewed and evidence-based gene/drug clinical practice guidelines indicating preemptive tPMt genotype-guided thiopurine-dose individualization to mitigate drug toxicity. Recently loss-of-function germline variants in NUDT15 have been identified as additional genetic determinants of thiopurine intolerance. MetHoDS: We used a taqMaMa genotyping Pcr assay NUDt15-MaMaPcr real time cod. BioMole 020 for detection of c.415 c-to-t transition (rs116855232) variant of the nucleoside diphosphate-linked moiety X motif 15 (NUDt15) gene on 7500 real-time Pcr System and cFX96 real-time Pcr System. Subsequently we used this assay in subject with thiopurine treatment. RESULTS: Homozygotes and heterozygotes condition were tested. We reported the ΔCt obtained from mimic homozygotes C/C (CtsC 22.31, CtaT 28.90 with ΔCt 6.59) and T/T (CtsT 22.75, CtaC 31.01 with ΔCt 8.26) and heterozygotes C/T conditions (CtsC 23.33, CtsT 23.72 with ΔCt 0.38). For the clinical validation of the techniques we analyzed a population of 31 thiopurine treated pediatric patients blood samples (13 male, 18 female). all 31 samples were tested with 31/31 (100%) homozygotes (31 cc). coNclUSioNS: the cFX96 real-time Pcr System is a suitable instrument to use in conjunction with the NUDt15-MaMaPcr real time to generate predictive information about genotyping assay of rs116855232, c415t polymorphism in children with thiopurine treatment.
evaluation of the polymorphism NUDt15c.415c>t real-time Pcr on the cFX96 real-time Pcr system and 7500 real-time Pcr system
Galliano I.;Alliaudi C.;Calvi C.;Montanari P.;Bergallo M.
2020-01-01
Abstract
BacKgroUND: thiopurine-related hematoxicity is clinically relevant and therapy-limiting in treatment of patients with acute lymphoblastic leukemia (ALL) or inflammatory bowel disease (IBD). For clinical implementation, the Clinical Pharmacogenetics Implementation Consortium (cPic) published peer-reviewed and evidence-based gene/drug clinical practice guidelines indicating preemptive tPMt genotype-guided thiopurine-dose individualization to mitigate drug toxicity. Recently loss-of-function germline variants in NUDT15 have been identified as additional genetic determinants of thiopurine intolerance. MetHoDS: We used a taqMaMa genotyping Pcr assay NUDt15-MaMaPcr real time cod. BioMole 020 for detection of c.415 c-to-t transition (rs116855232) variant of the nucleoside diphosphate-linked moiety X motif 15 (NUDt15) gene on 7500 real-time Pcr System and cFX96 real-time Pcr System. Subsequently we used this assay in subject with thiopurine treatment. RESULTS: Homozygotes and heterozygotes condition were tested. We reported the ΔCt obtained from mimic homozygotes C/C (CtsC 22.31, CtaT 28.90 with ΔCt 6.59) and T/T (CtsT 22.75, CtaC 31.01 with ΔCt 8.26) and heterozygotes C/T conditions (CtsC 23.33, CtsT 23.72 with ΔCt 0.38). For the clinical validation of the techniques we analyzed a population of 31 thiopurine treated pediatric patients blood samples (13 male, 18 female). all 31 samples were tested with 31/31 (100%) homozygotes (31 cc). coNclUSioNS: the cFX96 real-time Pcr System is a suitable instrument to use in conjunction with the NUDt15-MaMaPcr real time to generate predictive information about genotyping assay of rs116855232, c415t polymorphism in children with thiopurine treatment.
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