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Yanggu, South Korea

Kim H.,Seoul National University | Kang H.J.,Seoul National University | Kim H.J.,Seoul National University | Jang M.K.,Seoul National University | And 9 more authors.

Genetic polymorphisms are important factors in the effects and toxicity of chemotherapeutics. To analyze the pharmacogenetic and ethnic differences in chemotherapeutics, major genes implicated in the treatment of acute lymphoblastic leukemia (ALL) were analyzed. Eighteen loci of 16 genes in 100 patients with ALL were analyzed. The distribution of variant alleles were CYP3A4*1B (0%), CYP3A5*3 (0%), GSTM1 (21%), GSTP1 (21%), GSTT1 (16%), MDR1 exon 21 (77%), MDR1 exon 26 (61%), MTHFR 677 (63%), MTHFR 1298 (29%), NR3C1 1088 (0%), RFC1 80 (68%), TPMT combined genotype (7%), VDR intron 8 (11%), VDR FokI (83%), TYMS enhancer repeat (22%) and ITPA 94 (30%). The frequencies of single nucleotide polymorphisms (SNPs) of 10 loci were statistically different from those in Western Caucasians. Dose percents (actual/planned dose) or toxicity of mercaptopurine and methotrexate were not related to any SNPs. Event free survival (EFS) rate was lower in ITPA variants, and ITPA 94 AC/AA variant genotypes were the only independent risk factor for lower EFS in multivariate analysis, which was a different pharmacogenetic implication from Western studies. This study is the first pharmacogenetic study in Korean pediatric ALL. Our result suggests that there are other possible pharmacogenetic factors besides TPMT or ITPA polymorphisms which influence the metabolism of mercaptopurine in Asian populations. © 2012 Kim et al. Source

Park H.J.,YeBT Co. | Park H.J.,Seoul National University | Oh Y.,YeBT Co. | Kang H.J.,Seoul National University | And 6 more authors.
Tissue Antigens

A simple and accurate method for killer-cell immunoglobulin-like receptor (KIR) genotyping is developed using KIR gene-specific primer extension (GSPE) followed by bead array hybridization (GSPE method). After amplification of exons 4, 5, and 9, KIR GSPE and bead array hybridization were performed to verify the presence or absence of 16 KIR subfamilies. GSPE method was validated with natural killer/KIR reference panel I consisting of 48 cell types provided by 13th International Histocompatibility Working Group (IHWG) and genomic DNA from 17 peripheral blood cells, 8 cell lines, and 8 buccal cells. The results of reference panel from GSPE method were 100% concordant with the IHWG reference typing information. All genomic DNAs except reference panel were typed for KIR genes with sequence-specific primer methods and showed 100% identical typing results using this novel system. In addition, GSPE method can obtain results in 8 h from DNA with 10 ng genomic DNA in a 96-well-based assay format. © 2011 John Wiley & Sons A/S. Source

Kim M.S.,Seoul National University | Kang H.J.,Seoul National University | Park H.J.,YeBT Co. | Yook Y.-J.,Seoul National University | And 8 more authors.
Molecular Diagnosis and Therapy

Background: Busulfan is a key compound in myeloablative chemotherapy before hematopoietic stem-cell transplantation in children. Genetic polymorphisms of glutathione S-transferase (GST), which is involved in the metabolism of busulfan, have been implicated in interindividual variability in busulfan pharmacokinetics. Development of a rapid and simplified method for polygenic analysis of GST may facilitate large pharmacogenetic studies and clinical application of individualized busulfan dose adjustment. We previously introduced an effective PCR method for analyzing multiple genes using a small amount of DNA, termed 'TotalPlex amplification'. Objective: The aim of this study was to extend the application of the TotalPlex method to the specific GST gene families (A1, P1, M1, and T1) that are related to busulfan metabolism, and thereby facilitate pharmacogenetic analysis of GST polymorphisms. Methods: Seven genetic polymorphisms (GSTA1 promoter -52G>A, -69C>T, -567T>G, and -631T>G; GSTP1 313A>G; GSTM1 deletion; and GSTT1 deletion) were analyzed by multiplex PCR and genotyping, and the genotyping results from TotalPlex were verified with those from uniplex PCR. Results: Using five pairs of specific bulging-specific primers, seven specific gene fragments were successfully amplified by multiplex amplification coupled to a multiplexed bead array detection system, with a smaller amount of DNA and a shorter process time than is needed for the conventional approach. The genotypes of seven loci from 30 different genomic DNA samples derived using the multiplex system were consistent with the results of standard genotyping methods. Conclusion: Our multiplex system provides a fast, inexpensive, and accurate method of detecting multiple GST polymorphisms (GSTA1, GSTP1, GSTM1, and GSTT1). © 2011 Adis Data Information BV. All rights reserved. Source

Choi S.H.,Seoul Womens University | Taek Oh Y.,YeBT Co. | Kwon J.Y.,Seoul Womens University | Lee S.N.,Seoul Womens University | And 2 more authors.
Journal of Applied Biological Chemistry

A multiplex system was developed to assess detection of stacked genetically modified (GM) rice (LS28 × Cry1Ac) based on multiplex polymerase chain reaction (PCR) and liquid beadarray, and the accuracy of the system was analyzed. Standard and specific bulging specific (SBS) primers with standard primers were used to simultaneously detect multiple targets in stacked events of rice. Five sets of primers for the stacked events were applied to amplify their targets, and were separated distinctly in agarose gel. A liquid beadarray assay for the stacked GM rice was performed using the multiplex PCR products, followed by target biotinylation and hybridization between biotinylated-tagged target and anti-tagged bead. Fluorescent signals of the hybridized target sequences were detected by the Luminex system. The signaling patterns were analyzed by their mean fluorescent intensity (MFI) value. Results showed that liquid beadarrays with standard and SBS primers were in complete agreement with the PCR data, and detection of the different target elements was found to be very specific with no cross reaction among samples. Therefore, our detection system developed for stacked GM crop using multiplex PCR and liquid beadarray can be a useful and efficient system for screening and analyzing multiple transgenes in a single tube for qualitative analysis. Source

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