A direct isothermal amplification system adapted for rapid SNP genotyping of multifarious sample types

Publication date: 15 September 2018
Source:Biosensors and Bioelectronics, Volume 115
Author(s): Xiaonan Liu, Chao Zhang, Mengye Zhao, Kewu Liu, Hang Li, Ningning Li, Linlin Gao, Xuemin Yang, Ting Ma, Juanli Zhu, Wenli Hui, Kai Hua, Yali Cui
Genotyping of single nucleotide polymorphisms (SNPs) in point-of-care (POC) settings could be further improved through simplifying the treatment of samples. In this study, we devised an accurate, rapid and easy-to-use SNP detection system based on direct loop-mediated isothermal amplification (LAMP) without DNA extraction, known as Direct-LAMP. Samples from various sources (including whole blood, dried blood spot, buccal swab and saliva), treated with NaOH, can be used directly in amplification. The turnaround time was about 30 min from sample collection to provision of results. The accuracy was evaluated by assessing the polymorphisms of methylenetetrahydrofolate reductase (MTHFR) C677T and aldehyde dehydrogenase-2 (ALDH2) Glu504Lys, which are better known for their critical role in folate and ethanol metabolism, respectively. Completely consistent genotyping results reveal that Direct-LAMP is generally concordant with sequencing. This system can serve as a very promising platform in the fields of disease predisposition, drug metabolism and personalized medicine.

Graphical abstract

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