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Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions
Published on: September 25, 2018
Simultaneous detection of CYP3A5 and MDR1 polymorphisms based on the SNaPshot assay
Liang Li1, Chuan-Jiang Li, Yan-Jun Zhang
1School of Basic Medical Sciences, Southern Medical University, Guangzhou, PR China.
Background:
The 6986A>G polymorphism for CYP3A5 and the -129T>C, 1236C>T, 2677G>T/A, and 3435C>T polymorphisms for MDR1 are considered the major genetic factors affecting a range of drugs' metabolism and transport. Simultaneous genotyping of these five polymorphisms would be useful for estimating the therapeutic effects of their related drugs.
Subjects And Methods:
We have described a SNaPshot assay that can simultaneously detect all the five polymorphisms based on multiplex PCR and minisequencing reaction. A total of 168 unrelated Chinese DNA samples were used to establish and evaluate the assay.
Results:
The different genotypes of the five polymorphisms could be determined by peak retention time and colors. DNA sequencing was performed on samples randomly selected from each of the genotype groups detected by SNaPshot assay, and the results indicated 100% concordance.
Conclusion:
The SNaPshot assay for the CYP3A5 and MDR1 five polymorphisms detection was accurate, automated, and cost-effective.
