Evaluation of T-ARMS-PCR for identification of −130G>A in metformin transporter gene SLC47A2/MATE2 using saliva-derived DNA
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1
International Medical University, Malaysia
Background
Interindividual responses of metformin, a widely used drug for type 2 diabetes mellitus (T2D) have been reported in several studies. SLC47A2 gene that encodes a major transporter protein MATE2 is profoundly associated with the pharmacokinetic and pharmacodynamic variations of drugs including metformin. Of relevance, a single nucleotide polymorphism (SNP) known as −130G>A (rs12943590) is significantly associated with altered glycemic response in T2D patients with metformin. Hence, a reliable and efficient method is essential to identify −130G>A (rs12943590) in clinical laboratory for implementation of personalised medicine in future.
Methods
Tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR), a rapid and simple genotyping assay was developed to detect −130G>A (rs12943590). The concentration and purity of saliva-derived DNA were evaluated using a nanophotometer. The absorbance ratio of A260/280 and A260/230 was used to determine DNA quality and purity. Subsequently, the validated method was used to determine the genotype distribution and allele frequencies of −130G>A (rs12943590) in 150 of healthy volunteers.
Results
The assay demonstrated 100% concordance between the direct sequencing and T-ARMS PCR detection. The genotypes distribution of −130G>A namely; AA, AG and GG were 24.7%, 47.3% and 28.0%, respectively in accordance to the Hardy-Weinberg Equilibrium. The lowest detection limit of T-ARMS-PCR assay in this study was 40ng.
Conclusion
In summary, a diagnostic validation of T-ARMS PCR for detection of −130G>A (rs12943590) was successfully performed. T-ARMS-PCR is a rapid and ideal assay for genotype screening in clinical laboratory settings.
Acknowledgements
This research funded by International Medical University grant (BPI-1/14(16)2017).
Keywords:
type 2 diabetes mellitus,
T-ARMS-PCR,
SLC47A2/MATE2,
Single nucleotide polymorphism,
Metformin
Conference:
International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18)
“Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019.
Presentation Type:
Poster Presentation
Topic:
Metabolic diseases
Citation:
HASHIM
H,
Chee Chuan
C,
Teo Yong Xuan
E,
Siew Ching
H and
Pua Wee Kiet
J
(2019). Evaluation of T-ARMS-PCR for identification of −130G>A in metformin transporter gene SLC47A2/MATE2 using saliva-derived DNA.
Front. Pharmacol.
Conference Abstract:
International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18)
“Seizing Opportunities and Addressing Challenges of Precision Medicine”.
doi: 10.3389/conf.fphar.2018.63.00081
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Received:
28 Sep 2018;
Published Online:
17 Jan 2019.
*
Correspondence:
Dr. HAZWANIE HASHIM, International Medical University, Kuala Lumpur, Malaysia, hazwaniehashim@imu.edu.my