Development of an in-house method for genotyping of ABCB1 3435C>T and its association with dose, concentration of tacrolimus in renal transplant patients

  • Hoang Xuan Su Military Medical University
  • Nguyen Giang Hoa 103 Military Hospital
  • Ha Thanh Binh Quân chủng Hải Quân
  • Dinh Thi Thu Hang Military Medical Academy
  • Nguyen Huu Ben Military Medical Academy
  • Pham Quoc Toan 103 Military Hospital

Main Article Content

Keywords

ABCB1, PCR-RFLP, tacrolimus, renal transplant

Abstract

Objective: To develop an in-house method for genotyping of ABCB1 genetic polymorphism and analyze its impact on the concentration and dose of tacrolimus in renal transplant patients. Subject and method: A total of 100 patients after kidney transplant were treated with tacrolimus at the Department of Nephrology and Dialysis - 103 Military Hospital, Military Medical University. Blood samples were collected to identify ABCB1 genetic polymorphisms of and monitor Tacrolimus levels. Primer design and restriction enzyme selection for setting up the genotyping of the polymorphism at 3435C>T (rs1045642). Analyzing the relationship between ABCB1 genotypes obtained with doses and concentrations of TAC at the time points of follow up. Result: In this study, a fragment 244bp containing ABCB1 3435C>T (rs1045642) genetic polymorphism was amplified by PCR. Enzyme MboI recognize the PCR product cutting sequence which generates 2 fragments of 172bp and 68bp, respectively, for the C allele or CC homozygous genotype. When the polymorphism is the T allele, the MboI enzyme cuts the PCR product to create a fragment with the corresponding size of 244bp or homozygous genotype TT. The CT heterozygous genotype corresponds to fragments with sizes of 244, 172, 68bp, respectively. There was no significant difference between ABCB1 3435C>T (rs1045642) genotypes and concentrations and doses of tacrolimus at different time points investigated. Conclusion: We have established a simple and reliable method for genotyping ABCB1 3435C>T (rs1045642). No effect of the polymorphism ABCB1 3435C>T (rs1045642) has been found in relation to the dose and concentration of tacrolimus in patients after kidney transplantation.

Article Details

References

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