Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone
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| internalnotes | Bart, G., Heilig, M., LaForge, K. S., Pollak, L., Leal, S. M., Ott, J., & Kreek, M. J. (2004). Substantial attributable risk related to a functional mu-opioid receptor gene polymorphism in association with heroin addiction in central sweden. Molecular Psychiatry, 9(6), 547-549+542. doi:10.1038/sj.mp.4001504 Bergen, A. W., Kokoszka, J., Peterson, R., Long, J. C., Virkkunen, M., Linnoila, M., & Goldman, D. (1997). μ Opioid receptor gene variants: Lack of association with alcohol dependence. Molecular Psychiatry, 2(6), 490-494. doi:10.1038/sj.mp.4000331 Blievernicht, J. K., Schaeffeler, E., Klein, K., Eichelbaum, M., Schwab, M., & Zanger, U. M. (2007). MALDI-TOF mass spectrometry for multiplex genotyping of CYP2B6 single-nucleotide polymorphisms. Clinical Chemistry, 53(1), 24-33. doi:10.1373/clinchem.2006.074856 Bond, C., Laforge, K. S., Tian, M., Melia, D., Zhang, S., Borg, L., . . . Yu, L. (1998). Single-nucleotide polymorphism in the human mu opioid receptor gene alters β-endorphin binding and activity: Possible implications for opiate addiction. Proceedings of the National Academy of Sciences of the United States of America, 95(16), 9608-9613. doi:10.1073/pnas.95.16.9608 Bunten, H., Liang, W. J., Pounder, D. J., Seneviratne, C., & Osselton, D. (2010). OPRM1 and CYP2B6 gene variants as risk factors in methadone-related deaths. Clinical Pharmacology and Therapeutics, 88(3), 383-389. doi:10.1038/clpt.2010.127 Chamberlain, J. S., & Chamberlain, J. R. (1994). Optimization of multiplex PCRs. The Polymerase Chain Reaction, , 38-46. Retrieved from www.scopus.com Corkery, J. M., Schifano, F., Ghodse, A. H., & Oyefeso, A. (2004). The effects of methadone and its role in fatalities. Human Psychopharmacology, 19(8), 565-576. doi:10.1002/hup.630 Crettol, S., Déglon, J. -., Besson, J., Croquette-Krokkar, M., Gothuey, I., Hämmig, R., . . . Eap, C. B. (2005). Methadone enantiomer plasma levels, CYP2B6, CYP2C19, and CYP2C9 genotypes, and response to treatment. Clinical Pharmacology and Therapeutics, 78(6), 593-604. doi:10.1016/j.clpt.2005.08.011 Eap, C. B., Crettol, S., Rougier, J. -., Schläpfer, J., Sintra Grilo, L., Déglon, J. -., . . . Abriel, H. (2007). Stereoselective block of hERG channel by (S)-methadone and QT interval prolongation in CYP2B6 slow metabolizers. Clinical Pharmacology and Therapeutics, 81(5), 719-728. doi:10.1038/sj.clpt.6100120 Foster, D. J. R., Somogyi, A. A., & Bochner, F. (1999). Methadone N-demethylation in human liver microsomes: Lack of stereoselectivity and involvement of CYP3A4. British Journal of Clinical Pharmacology, 47(4), 403-412. doi:10.1046/j.1365-2125.1999.00921.x Gerber, J. G., Rhodes, R. J., & Gal, J. (2004). Stereoselective metabolism of methadone N-demethylation by cytochrome P4502B6 and 2C19. Chirality, 16(1), 36-44. doi:10.1002/chir.10303 Giacomini, K. M., Brett, C. M., Altman, R. B., Benowitz, N. L., Dolan, M. E., Flockhart, D. A., . . . Zineh, I. (2007). The pharmacogenetics research network: From SNP discovery to clinical drug response. Clinical Pharmacology and Therapeutics, 81(3), 328-345. doi:10.1038/sj.clpt.6100087 Henegariu, O., Heerema, N. A., Dlouhy, S. R., Vance, G. H., & Vogt, P. H. (1997). Multiplex PCR: Critical parameters and step-by-step protocol. BioTechniques, 23(3), 504-511. doi:10.2144/97233rr01 Ide, S., Kobayashi, H., Ujike, H., Ozaki, N., Sekine, Y., Inada, T., . . . Sora, I. (2006). Linkage disequilibrium and association with methamphetamine dependence/psychosis of μ-opioid receptor gene polymorphisms. Pharmacogenomics Journal, 6(3), 179-188. doi:10.1038/sj.tpj.6500355 Ingelman-Sundberg, M., Sim, S. C., Gomez, A., & Rodriguez-Antona, C. (2007). Influence of cytochrome P450 polymorphisms on drug therapies: Pharmacogenetic, pharmacoepigenetic and clinical aspects. Pharmacology and Therapeutics, 116(3), 496-526. doi:10.1016/j.pharmthera.2007.09.004 Jacob, R. M., Johnstone, E. C., Neville, M. J., & Walton, R. T. (2004). Identification of CYP2B6 sequence variants by use of multiplex PCR with allele-specific genotyping. Clinical Chemistry, 50(8), 1372-1377. doi:10.1373/clinchem.2004.031708 Jannetto, P. J., Wong, S. H., Gock, S. B., Laleli-Sahin, E., Schur, B. C., & Jentzen, J. M. (2002). Pharmacogenomics as molecular autopsy for postmortem forensic toxicology: Genotyping cytochrome P450 2D6 for oxycodone cases. Journal of Analytical Toxicology, 26(7), 438-447. doi:10.1093/jat/26.7.438 Kharasch, E. D., Hoffer, C., Whittington, D., & Sheffels, P. (2004). Role of hepatic and intestinal cytochrome P450 3A and 2B6 in the metabolism, disposition, and miotic effects of methadone. Clinical Pharmacology and Therapeutics, 76(3), 250-269. doi:10.1016/j.clpt.2004.05.003 Klepstad, P., Dale, O., Skorpen, F., Borchgrevink, P. C., & Kaasa, S. (2005). Genetic variability and clinical efficacy of morphine. Acta Anaesthesiologica Scandinavica, 49(7), 902-908. doi:10.1111/j.1399-6576.2005.00772.x Li, T., Liu, X., Zhu, Z. H., Zhao, J., Hu, X., Sham, P. C., & Collier, D. A. (2000). Association analysis of polymorphisms in the μ opioid gene and heroin abuse in chinese subjects. Addiction Biology, 5(2), 181-186. doi:10.1080/13556210050003775 Lötsch, J., Zimmermann, M., Darimont, J., Marx, C., Dudziak, R., Skarke, C., & Geisslinger, G. (2002). Does the A118G polymorphism at the μ-opioid receptor gene protect against morphine-6-glucuronide toxicity? Anesthesiology, 97(4), 814-819. doi:10.1097/00000542-200210000-00011 Mathiesen, S., Stenz Justesen, U., Von L̈ttichau, H. -., & Eg Hansen, A. -. (2006). Genotyping of CYP2B6 and therapeutic drug monitoring in an HIV-infected patient with high efavirenz plasma concentrations and severe CNS side-effects. Scandinavian Journal of Infectious Diseases, 38(8), 733-735. doi:10.1080/00365540500504109 Maxwell, J. C., Pullum, T. W., & Tannert, K. (2005). Deaths of clients in methadone treatment in texas: 1994-2002. Drug and Alcohol Dependence, 78(1), 73-81. doi:10.1016/j.drugalcdep.2004.09.006 Mills, R. E., Luttig, C. T., Larkins, C. E., Beauchamp, A., Tsui, C., Pittard, W. S., & Devine, S. E. (2006). An initial map of insertion and deletion (INDEL) variation in the human genome. Genome Research, 16(9), 1182-1190. doi:10.1101/gr.4565806 Moffat, A. C., Osselton, M. D., & Widdop, B. (0000). Clarke's Analysis of Drugs and Poisons: In Pharmaceuticals, Body Fluids and Post-Mortem Material, 2003, 821. Retrieved from www.scopus.com Nettleton, R. T., Ransom, T. A., Abraham, S. L., Nelson, C. S., & Olsen, G. D. (2007). Methadone-induced respiratory depression in the neonatal guinea pig. Pediatric Pulmonology, 42(12), 1134-1143. doi:10.1002/ppul.20707 Osselton, M. D., Blackmore, R. C., King, L. A., & Moffat, A. C. (1984). Poisoning-associated deaths for england and wales between 1973 and 1980. Human & Experimental Toxicology, 3(3), 201-221. doi:10.1177/096032718400300305 Powers, V., Ward, J., & Gompels, M. (2009). CYP2B6 G516T genotyping in a UK cohort of HIV-positive patients: Polymorphism frequency and influence on efavirenz discontinuation. HIV Medicine, 10(8), 520-523. doi:10.1111/j.1468-1293.2009.00718.x Ross, J. R., Rutter, D., Welsh, K., Joel, S. P., Goller, K., Wells, A. U., . . . Riley, J. (2005). Clinical response to morphine in cancer patients and genetic variation in candidate genes. Pharmacogenomics Journal, 5(5), 324-336. doi:10.1038/sj.tpj.6500327 Rotger, M., & Telenti, A. (2008). Optimizing efavirenz treatment: CYP2B6 genotyping or therapeutic drug monitoring? European Journal of Clinical Pharmacology, 64(4), 335-336. doi:10.1007/s00228-007-0440-z Schoske, R., Vallone, P. M., Ruitberg, C. M., & Butler, J. M. (2003). Multiplex PCR design strategy used for the simultaneous amplification of 10 y chromosome short tandem repeat (STR) loci. Analytical and Bioanalytical Chemistry, 375(3), 333-343. doi:10.1007/s00216-002-1683-2 Wang, J. -., & DeVane, C. L. (2003). Involvement of CYP3A4, CYP2C8, and CYP2D6 in the metabolism of (R)- and (S)-methadone in vitro. Drug Metabolism and Disposition, 31(6), 742-747. doi:10.1124/dmd.31.6.742 White Sr., R. M., & Wong, S. H. (2005). Pharmacogenomics and its applications. MLO: Medical Laboratory Observer, 37(3), 20-27. Retrieved from www.scopus.com Wolff, K. (2002). Characterization of methadone overdose: Clinical considerations and the scientific evidence. Therapeutic Drug Monitoring, 24(4), 457-470. doi:10.1097/00007691-200208000-00001 Wong, S. H., Wagner, M. A., Jentzen, J. M., Schur, C., Bjerke, J., Gock, S. B., & Chang, C. -. (2003). Pharmacogenomics as an aspect of molecular autopsy for forensic Pathology/Toxicology: Does genotyping CYP 2D6 serve as an adjunct for certifying methadone toxicity? Journal of Forensic Sciences, 48(6), 1406-1415. doi:10.1520/jfs2002392 Zhou, S. -., Liu, J. -., & Chowbay, B. (2009). Polymorphism of human cytochrome P450 enzymes and its clinical impact. Drug Metabolism Reviews, 41(2), 89-295. doi:10.1080/03602530902843483 |
| originalfilename | 5314-01-FH02-FPSK-14-00785.jpg |
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| spelling | 11117 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=11117 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection407072 Restricted Document Article Journal UniSZA Unisza unisza image/jpeg inches 96 96 1419 63 63 773 2014-07-02 14:46:39 1419x773 5314-01-FH02-FPSK-14-00785.jpg UniSZA Private Access Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone International Journal of Pharmacy and Pharmaceutical Sciences Objective: Cytochrome P450 2B6 (CYP2B6) is involved in the metabolism of several therapeutically important drugs and abused intoxicants including methadone. The preferential binding target for methadone is the μ opioid receptor OPRM1. Various SNPs in CYP2B6 and OPRM1 may contribute to clinical outcomes in methadone maintenance therapy. The aim of the present study was to develop a consistent and robust assay to identify such genotypic variants. Methods: We randomly selected patients in the Malay population who were addicted to heroin and had already commenced methadone therapy. We developed a two-step, multiplexed, nested PCR method to simultaneously identify 26 SNPs in CYP2B6 and OPRM1. Three amplicons covering seven regions of interest were amplified in the first PCR step. These amplicons were then used as templates for 11 sets of reactions in the second PCR step. Results: We successfully amplified DNA fragments containing 26 SNPs of interest from the DNA samples provided by patients receiving methadone therapy: C534T, G779A, G13076A, A454G, G24A, G820A, G942A, G31A, G438A, T106C, C440G, T397A, T15708C, T480C, C17T, G794A, A13072G, A76T, A62T, C64T, A136G, C15618T, G691C, G15631T, T802C, and A118G. Conclusion: The PCR-based method was sensitive and specific and is the first nested, allele-specific, multiplexed PCR method reported for the simultaneous detection of all 26 clinically important SNPs of CYP2B6 and OPMR1. 6 3 215-222 Bart, G., Heilig, M., LaForge, K. S., Pollak, L., Leal, S. M., Ott, J., & Kreek, M. J. (2004). Substantial attributable risk related to a functional mu-opioid receptor gene polymorphism in association with heroin addiction in central sweden. Molecular Psychiatry, 9(6), 547-549+542. doi:10.1038/sj.mp.4001504 Bergen, A. W., Kokoszka, J., Peterson, R., Long, J. C., Virkkunen, M., Linnoila, M., & Goldman, D. (1997). μ Opioid receptor gene variants: Lack of association with alcohol dependence. Molecular Psychiatry, 2(6), 490-494. doi:10.1038/sj.mp.4000331 Blievernicht, J. K., Schaeffeler, E., Klein, K., Eichelbaum, M., Schwab, M., & Zanger, U. M. (2007). MALDI-TOF mass spectrometry for multiplex genotyping of CYP2B6 single-nucleotide polymorphisms. Clinical Chemistry, 53(1), 24-33. doi:10.1373/clinchem.2006.074856 Bond, C., Laforge, K. S., Tian, M., Melia, D., Zhang, S., Borg, L., . . . Yu, L. (1998). Single-nucleotide polymorphism in the human mu opioid receptor gene alters β-endorphin binding and activity: Possible implications for opiate addiction. Proceedings of the National Academy of Sciences of the United States of America, 95(16), 9608-9613. doi:10.1073/pnas.95.16.9608 Bunten, H., Liang, W. J., Pounder, D. J., Seneviratne, C., & Osselton, D. (2010). OPRM1 and CYP2B6 gene variants as risk factors in methadone-related deaths. Clinical Pharmacology and Therapeutics, 88(3), 383-389. doi:10.1038/clpt.2010.127 Chamberlain, J. S., & Chamberlain, J. R. (1994). Optimization of multiplex PCRs. The Polymerase Chain Reaction, , 38-46. Retrieved from www.scopus.com Corkery, J. M., Schifano, F., Ghodse, A. H., & Oyefeso, A. (2004). The effects of methadone and its role in fatalities. Human Psychopharmacology, 19(8), 565-576. doi:10.1002/hup.630 Crettol, S., Déglon, J. -., Besson, J., Croquette-Krokkar, M., Gothuey, I., Hämmig, R., . . . Eap, C. B. (2005). Methadone enantiomer plasma levels, CYP2B6, CYP2C19, and CYP2C9 genotypes, and response to treatment. Clinical Pharmacology and Therapeutics, 78(6), 593-604. doi:10.1016/j.clpt.2005.08.011 Eap, C. B., Crettol, S., Rougier, J. -., Schläpfer, J., Sintra Grilo, L., Déglon, J. -., . . . Abriel, H. (2007). Stereoselective block of hERG channel by (S)-methadone and QT interval prolongation in CYP2B6 slow metabolizers. Clinical Pharmacology and Therapeutics, 81(5), 719-728. doi:10.1038/sj.clpt.6100120 Foster, D. J. R., Somogyi, A. A., & Bochner, F. (1999). Methadone N-demethylation in human liver microsomes: Lack of stereoselectivity and involvement of CYP3A4. British Journal of Clinical Pharmacology, 47(4), 403-412. doi:10.1046/j.1365-2125.1999.00921.x Gerber, J. G., Rhodes, R. J., & Gal, J. (2004). Stereoselective metabolism of methadone N-demethylation by cytochrome P4502B6 and 2C19. Chirality, 16(1), 36-44. doi:10.1002/chir.10303 Giacomini, K. M., Brett, C. M., Altman, R. B., Benowitz, N. L., Dolan, M. E., Flockhart, D. A., . . . Zineh, I. (2007). The pharmacogenetics research network: From SNP discovery to clinical drug response. Clinical Pharmacology and Therapeutics, 81(3), 328-345. doi:10.1038/sj.clpt.6100087 Henegariu, O., Heerema, N. A., Dlouhy, S. R., Vance, G. H., & Vogt, P. H. (1997). Multiplex PCR: Critical parameters and step-by-step protocol. BioTechniques, 23(3), 504-511. doi:10.2144/97233rr01 Ide, S., Kobayashi, H., Ujike, H., Ozaki, N., Sekine, Y., Inada, T., . . . Sora, I. (2006). Linkage disequilibrium and association with methamphetamine dependence/psychosis of μ-opioid receptor gene polymorphisms. Pharmacogenomics Journal, 6(3), 179-188. doi:10.1038/sj.tpj.6500355 Ingelman-Sundberg, M., Sim, S. C., Gomez, A., & Rodriguez-Antona, C. (2007). Influence of cytochrome P450 polymorphisms on drug therapies: Pharmacogenetic, pharmacoepigenetic and clinical aspects. Pharmacology and Therapeutics, 116(3), 496-526. doi:10.1016/j.pharmthera.2007.09.004 Jacob, R. M., Johnstone, E. C., Neville, M. J., & Walton, R. T. (2004). Identification of CYP2B6 sequence variants by use of multiplex PCR with allele-specific genotyping. Clinical Chemistry, 50(8), 1372-1377. doi:10.1373/clinchem.2004.031708 Jannetto, P. J., Wong, S. H., Gock, S. B., Laleli-Sahin, E., Schur, B. C., & Jentzen, J. M. (2002). Pharmacogenomics as molecular autopsy for postmortem forensic toxicology: Genotyping cytochrome P450 2D6 for oxycodone cases. Journal of Analytical Toxicology, 26(7), 438-447. doi:10.1093/jat/26.7.438 Kharasch, E. D., Hoffer, C., Whittington, D., & Sheffels, P. (2004). Role of hepatic and intestinal cytochrome P450 3A and 2B6 in the metabolism, disposition, and miotic effects of methadone. Clinical Pharmacology and Therapeutics, 76(3), 250-269. doi:10.1016/j.clpt.2004.05.003 Klepstad, P., Dale, O., Skorpen, F., Borchgrevink, P. C., & Kaasa, S. (2005). Genetic variability and clinical efficacy of morphine. Acta Anaesthesiologica Scandinavica, 49(7), 902-908. doi:10.1111/j.1399-6576.2005.00772.x Li, T., Liu, X., Zhu, Z. H., Zhao, J., Hu, X., Sham, P. C., & Collier, D. A. (2000). Association analysis of polymorphisms in the μ opioid gene and heroin abuse in chinese subjects. Addiction Biology, 5(2), 181-186. doi:10.1080/13556210050003775 Lötsch, J., Zimmermann, M., Darimont, J., Marx, C., Dudziak, R., Skarke, C., & Geisslinger, G. (2002). Does the A118G polymorphism at the μ-opioid receptor gene protect against morphine-6-glucuronide toxicity? Anesthesiology, 97(4), 814-819. doi:10.1097/00000542-200210000-00011 Mathiesen, S., Stenz Justesen, U., Von L̈ttichau, H. -., & Eg Hansen, A. -. (2006). Genotyping of CYP2B6 and therapeutic drug monitoring in an HIV-infected patient with high efavirenz plasma concentrations and severe CNS side-effects. Scandinavian Journal of Infectious Diseases, 38(8), 733-735. doi:10.1080/00365540500504109 Maxwell, J. C., Pullum, T. W., & Tannert, K. (2005). Deaths of clients in methadone treatment in texas: 1994-2002. Drug and Alcohol Dependence, 78(1), 73-81. doi:10.1016/j.drugalcdep.2004.09.006 Mills, R. E., Luttig, C. T., Larkins, C. E., Beauchamp, A., Tsui, C., Pittard, W. S., & Devine, S. E. (2006). An initial map of insertion and deletion (INDEL) variation in the human genome. Genome Research, 16(9), 1182-1190. doi:10.1101/gr.4565806 Moffat, A. C., Osselton, M. D., & Widdop, B. (0000). Clarke's Analysis of Drugs and Poisons: In Pharmaceuticals, Body Fluids and Post-Mortem Material, 2003, 821. Retrieved from www.scopus.com Nettleton, R. T., Ransom, T. A., Abraham, S. L., Nelson, C. S., & Olsen, G. D. (2007). Methadone-induced respiratory depression in the neonatal guinea pig. Pediatric Pulmonology, 42(12), 1134-1143. doi:10.1002/ppul.20707 Osselton, M. D., Blackmore, R. C., King, L. A., & Moffat, A. C. (1984). Poisoning-associated deaths for england and wales between 1973 and 1980. Human & Experimental Toxicology, 3(3), 201-221. doi:10.1177/096032718400300305 Powers, V., Ward, J., & Gompels, M. (2009). CYP2B6 G516T genotyping in a UK cohort of HIV-positive patients: Polymorphism frequency and influence on efavirenz discontinuation. HIV Medicine, 10(8), 520-523. doi:10.1111/j.1468-1293.2009.00718.x Ross, J. R., Rutter, D., Welsh, K., Joel, S. P., Goller, K., Wells, A. U., . . . Riley, J. (2005). Clinical response to morphine in cancer patients and genetic variation in candidate genes. Pharmacogenomics Journal, 5(5), 324-336. doi:10.1038/sj.tpj.6500327 Rotger, M., & Telenti, A. (2008). Optimizing efavirenz treatment: CYP2B6 genotyping or therapeutic drug monitoring? European Journal of Clinical Pharmacology, 64(4), 335-336. doi:10.1007/s00228-007-0440-z Schoske, R., Vallone, P. M., Ruitberg, C. M., & Butler, J. M. (2003). Multiplex PCR design strategy used for the simultaneous amplification of 10 y chromosome short tandem repeat (STR) loci. Analytical and Bioanalytical Chemistry, 375(3), 333-343. doi:10.1007/s00216-002-1683-2 Wang, J. -., & DeVane, C. L. (2003). Involvement of CYP3A4, CYP2C8, and CYP2D6 in the metabolism of (R)- and (S)-methadone in vitro. Drug Metabolism and Disposition, 31(6), 742-747. doi:10.1124/dmd.31.6.742 White Sr., R. M., & Wong, S. H. (2005). Pharmacogenomics and its applications. MLO: Medical Laboratory Observer, 37(3), 20-27. Retrieved from www.scopus.com Wolff, K. (2002). Characterization of methadone overdose: Clinical considerations and the scientific evidence. Therapeutic Drug Monitoring, 24(4), 457-470. doi:10.1097/00007691-200208000-00001 Wong, S. H., Wagner, M. A., Jentzen, J. M., Schur, C., Bjerke, J., Gock, S. B., & Chang, C. -. (2003). Pharmacogenomics as an aspect of molecular autopsy for forensic Pathology/Toxicology: Does genotyping CYP 2D6 serve as an adjunct for certifying methadone toxicity? Journal of Forensic Sciences, 48(6), 1406-1415. doi:10.1520/jfs2002392 Zhou, S. -., Liu, J. -., & Chowbay, B. (2009). Polymorphism of human cytochrome P450 enzymes and its clinical impact. Drug Metabolism Reviews, 41(2), 89-295. doi:10.1080/03602530902843483 |
| spellingShingle | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| summary | Objective: Cytochrome P450 2B6 (CYP2B6) is involved in the metabolism of several therapeutically important drugs and abused intoxicants including methadone. The preferential binding target for methadone is the μ opioid receptor OPRM1. Various SNPs in CYP2B6 and OPRM1 may contribute to clinical outcomes in methadone maintenance therapy. The aim of the present study was to develop a consistent and robust assay to identify such genotypic variants. Methods: We randomly selected patients in the Malay population who were addicted to heroin and had already commenced methadone therapy. We developed a two-step, multiplexed, nested PCR method to simultaneously identify 26 SNPs in CYP2B6 and OPRM1. Three amplicons covering seven regions of interest were amplified in the first PCR step. These amplicons were then used as templates for 11 sets of reactions in the second PCR step. Results: We successfully amplified DNA fragments containing 26 SNPs of interest from the DNA samples provided by patients receiving methadone therapy: C534T, G779A, G13076A, A454G, G24A, G820A, G942A, G31A, G438A, T106C, C440G, T397A, T15708C, T480C, C17T, G794A, A13072G, A76T, A62T, C64T, A136G, C15618T, G691C, G15631T, T802C, and A118G. Conclusion: The PCR-based method was sensitive and specific and is the first nested, allele-specific, multiplexed PCR method reported for the simultaneous detection of all 26 clinically important SNPs of CYP2B6 and OPMR1. |
| title | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| title_full | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| title_fullStr | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| title_full_unstemmed | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| title_short | Simultaneous detection of multiple polymorphisms in CYP2B6 and OPMR1 by nested PCR: A good approach for patients on methadone |
| title_sort | simultaneous detection of multiple polymorphisms in cyp2b6 and opmr1 by nested pcr: a good approach for patients on methadone |