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Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease

  • Authors:
    • Rie Osaki
    • Hirotsugu Imaeda
    • Hiromitsu Ban
    • Tomoki Aomatsu
    • Shigeki Bamba
    • Tomoyuki Tsujikawa
    • Masaya Sasaki
    • Yoshihide Fujiyama
    • Akira Andoh
  • View Affiliations / Copyright

    Affiliations: Department of Medicine, Graduate School of Medicine, Shiga University of Medical Science, Seta Tukinowa, Otsu 520-2192, Japan, Division of Mucosal Immu­nology, Graduate School of Medicine, Shiga University of Medical Science, Seta Tukinowa, Otsu 520-2192, Japan
  • Pages: 783-786
    |
    Published online on: June 16, 2011
       https://doi.org/10.3892/etm.2011.287
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Abstract

Thiopurine drugs are the most common drugs used to maintain clinical remission in inflammatory bowel disease (IBD). Three single-nucleotide polymorphisms (SNPs), TPMT A719G (rs1142345), inosine triphosphate pyrophosphatase (ITPase) C94A (rs1127354) and multidrug resistance protein 4 MRP4 G2269A (rs3765534), have been reported to account for heightened sensitivity to thiopurine drugs in the Japanese population. We investigated the usefulness of the TaqMan® PCR assay (Applied Biosystems) for the rapid detection of these SNPs to improve the safety of thiopurine therapy. We enrolled 44 healthy volunteers and 235 IBD patients. Genotyping of the SNPs was performed using Custom TaqMan SNP genotyping assays, direct sequencing and PCR-RFLP. Genotyping for MRP4 G2269A by the TaqMan PCR assay was successfully achieved in all samples. Comparison with our previous data using direct sequencing indicated one discordant result, and re-sequencing showed that the TaqMan PCR assay was correct. The overall accuracy of the TaqMan assay for MRP4 G2269A was 100%. The TaqMan PCR genotyping for TPMT A719G and ITPase C94A was successfully performed in all samples. The results of TPMT A719G by the TaqMan assay were identical with those of PCR-RFLP. In ITPase C94A, a comparison of the TaqMan assay and PCR-RFLP yielded 12 discordant results, and direct sequencing showed that the TaqMan PCR assay was correct. The allelic frequency determined by the TaqMan assay was 0.145 for MRP4 G2269A, 0.009 for TPMT A719G and 0.121 for ITPase C94A, respectively. In conclusion, the TaqMan® PCR assay is useful for genotyping of SNPs responsible for thiopurine sensitivity in Japanese IBD patients.
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Copy and paste a formatted citation
Spandidos Publications style
Osaki R, Imaeda H, Ban H, Aomatsu T, Bamba S, Tsujikawa T, Sasaki M, Fujiyama Y and Andoh A: Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease. Exp Ther Med 2: 783-786, 2011.
APA
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T. ... Andoh, A. (2011). Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease. Experimental and Therapeutic Medicine, 2, 783-786. https://doi.org/10.3892/etm.2011.287
MLA
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T., Sasaki, M., Fujiyama, Y., Andoh, A."Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease". Experimental and Therapeutic Medicine 2.5 (2011): 783-786.
Chicago
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T., Sasaki, M., Fujiyama, Y., Andoh, A."Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease". Experimental and Therapeutic Medicine 2, no. 5 (2011): 783-786. https://doi.org/10.3892/etm.2011.287
Copy and paste a formatted citation
x
Spandidos Publications style
Osaki R, Imaeda H, Ban H, Aomatsu T, Bamba S, Tsujikawa T, Sasaki M, Fujiyama Y and Andoh A: Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease. Exp Ther Med 2: 783-786, 2011.
APA
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T. ... Andoh, A. (2011). Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease. Experimental and Therapeutic Medicine, 2, 783-786. https://doi.org/10.3892/etm.2011.287
MLA
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T., Sasaki, M., Fujiyama, Y., Andoh, A."Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease". Experimental and Therapeutic Medicine 2.5 (2011): 783-786.
Chicago
Osaki, R., Imaeda, H., Ban, H., Aomatsu, T., Bamba, S., Tsujikawa, T., Sasaki, M., Fujiyama, Y., Andoh, A."Accuracy of genotyping using the TaqMan PCR assay for single nucleotide polymorphisms responsible for thiopurine sensitivity in Japanese patients with inflammatory bowel disease". Experimental and Therapeutic Medicine 2, no. 5 (2011): 783-786. https://doi.org/10.3892/etm.2011.287
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