RSS-Feed abonnieren
PDF herunterladen
DOI: 10.1055/s-0043-1768948
A Simple, Rapid, and Cost-Effective PCR Procedure for Detection of NUDT15 Gene Variants in Vietnamese Patients with Acute Lymphoblastic Leukemia
Funding This study was financially supported in part by University of Medicine and Pharmacy at Ho Chi Minh City.
Abstract
Objective The NUDT15 variants impact thiopurine dose selection in acute lymphoblastic leukemia patients. The ability to rapidly detect variants is important in clinical practice. This study aims to develop a simple polymerase chain reaction (PCR) procedure for detecting NUDT15 variants in Vietnamese patients.
Materials and Methods Sanger sequencing was used to determine NUDT15 variants from 200 patients. We designed primers and optimized the PCR procedure for detection of wild-type and variant alleles and compared with Sanger sequencing results.
Results The inserted variant c.55_56insGAGTCG was detected by differences in size through conventional PCR. The tetra-primer amplification refractory mutation system PCR was successful in detecting two variations, c.52G > A and c.415C > T. The sensitivity and specificity of PCR procedure achieved 100% when compared to 200 Sanger sequencing results.
Conclusion Our PCR procedure is suitable for replacing Sanger sequencing to detect the NUDT15 variants in clinical setting.
* These authors contributed equally to this work.
Publikationsverlauf
Eingereicht: 04. März 2023
Angenommen: 05. April 2023
Artikel online veröffentlicht:
13. Juni 2023
© 2023. The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
-
References
- 1 Stelljes M, David IM. Acute Lymphoblastic Leukemia in Adults. The EBMT Handbook; 2019: 531-538
- 2 Haylett WJ. The relationship of genetics, nursing practice, and informatics tools in 6-mercaptopurine dosing in pediatric oncology [Formula: see text]. J Pediatr Oncol Nurs 2017; 34 (05) 342-346
- 3 Relling MV, Schwab M, Whirl-Carrillo M. et al. Clinical pharmacogenetics implementation consortium guideline for thiopurine dosing based on TPMT and NUDT 15 genotypes: 2018 update. Clin Pharmacol Ther 2019; 105 (05) 1095-1105
- 4 Rudin S, Marable M, Huang RS. The promise of pharmacogenomics in reducing toxicity during acute lymphoblastic leukemia maintenance treatment. Genomics Proteomics Bioinformatics 2017; 15 (02) 82-93
- 5 Mao X, Yin R, Sun G. et al. Effects of TPMT, NUDT15, and ITPA genetic variants on 6-mercaptopurine toxicity for pediatric patients with acute lymphoblastic leukemia in Yunnan of China. Front Pediatr 2021; 9: 719803
- 6 Sutiman N, Chen S, Ling KL. et al. Predictive role of NUDT15 variants on thiopurine-induced myelotoxicity in Asian inflammatory bowel disease patients. Pharmacogenomics 2018; 19 (01) 31-43
- 7 Yang JJ, Landier W, Yang W. et al. Inherited NUDT15 variant is a genetic determinant of mercaptopurine intolerance in children with acute lymphoblastic leukemia. J Clin Oncol 2015; 33 (11) 1235-1242
- 8 Moriyama T, Nishii R, Perez-Andreu V. et al. NUDT15 polymorphisms alter thiopurine metabolism and hematopoietic toxicity. Nat Genet 2016; 48 (04) 367-373
- 9 Medrano RFV, de Oliveira CA. Guidelines for the tetra-primer ARMS-PCR technique development. Mol Biotechnol 2014; 56 (07) 599-608
- 10 Dewi AL, Paramita DK, Fachiroh J. Development of tetra-primer amplification refractory mutation system (ARMS) PCR for detection of CHRNA3 rs8040868. Indones Biomed J 2021; 13: 192-200
- 11 Ahlawat S, Sharma R, Maitra A, Roy M, Tantia MS. Designing, optimization and validation of tetra-primer ARMS PCR protocol for genotyping mutations in caprine Fec genes. Meta Gene 2014; 2: 439-449
- 12 Nghia H, Than HH, Dong CV. et al. The effects of NUDT15 and TPMT variants on mercaptopurine treatment in Vietnamese pediatric acute lymphoblastic leukemia patients. Pediatr Hematol Oncol 2022; 39 (06) 561-570
- 13 Grover N, Bhatia P, Kumar A. et al. TPMT and NUDT15 polymorphisms in thiopurine induced leucopenia in inflammatory bowel disease: a prospective study from India. BMC Gastroenterol 2021; 21 (01) 327
- 14 Ho CC, Fong WY, Lee YH, Poon WT. Novel tetra-primer ARMS-PCR assays for thiopurine intolerance susceptibility mutations NUDT15 c.415C>T and TPMT c.719A>G (TPMT* 3C) in East Asians. Genes (Basel) 2017; 8 (10) 285-293
- 15 González-Lama Y, Gisbert JP. Monitoring thiopurine metabolites in inflammatory bowel disease. Frontline Gastroenterol 2016; 7 (04) 301-307