Citation: SHENG Nan, YUE Hui-Jie, PANG Shu-Yun, QI Xie-Min, ZHANG Yan-Jie, WU Yan-Zi, SONG Qin-Xin, ZOU Bing-Jie, ZHOU Guo-Hua. Visualized Detection of Aldehyde Dehydrogenase 2 Gene Polymorphism by Serial Invasive Reaction Coupled with Gold Nanoparticle Probe Assembling[J]. Chinese Journal of Analytical Chemistry, ;2021, 49(1): 42-49. doi: 10.19756/j.issn.0253-3820.201234 shu

Visualized Detection of Aldehyde Dehydrogenase 2 Gene Polymorphism by Serial Invasive Reaction Coupled with Gold Nanoparticle Probe Assembling

SHENG Nan ¹ ,
YUE Hui-Jie ¹ ,
PANG Shu-Yun ¹ ,
QI Xie-Min ¹ ,
ZHANG Yan-Jie ¹ ,
WU Yan-Zi ¹ ,
SONG Qin-Xin ^1,2 ,
ZOU Bing-Jie ^1,3,, ,
ZHOU Guo-Hua ^1,3,4,,

1.
Department of Pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing 210002, China;
2.
Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing 210009, China;
3.
Department of Chemistry, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210093, China;
4.
The First School of Clinical Medicine, Southern Medical University, Nanjing 210002, China

Corresponding author: ZOU Bing-Jie, ZHOU Guo-Hua,
Received Date: 26 April 2020
Revised Date: 27 October 2020

Fund Project: Supported by the National Natural Science Foundation of China (Nos.61871403, 81703474, 81673390), the Jiangsu Provincial Medical Youth Talent Program (No.QNRC2016889) and the Natural Science Foundation of Jiangsu Province (No.BK20180292).

Conventional assays for identifying polymorphism of aldehyde dehydrogenase 2 (ALDH2) gene (rs671) are based on electrophoresis, fluorescence-labeled probes or sequencing, which are laborious and expensive. Herein, a colorimetric method was proposed for detection of ALDH2 genotype by PCR-based serial invasive reaction coupled with gold nanoparticle probe (GNP) assembling, in which the target DNAs were firstly amplified by PCR, and then the alleles on the amplicons were identified by serial invasive reaction, and finally the genotyping results could be read out with naked eyes by observing the color changes caused by gold nanoparticle assembling. If the allele-specific invasive probe matched the target sequence, the hairpin probe could be cleaved into two parts by Flap endonuclease 1 (FEN1). As a result, the GNPs could not be aggregated by the hairpin probe, keeping the solution red. Otherwise, the intact hairpin probe caused the aggregation of GNPs, leading to the color change from red to purple or clear. Therefore, the genotyping result could be read out with naked eyes by observing the color changes of the reaction. As low as 80 pg of genomic DNA could be accurately genotyped, and the genotyping results of 207 clinical samples were consisted with that of pyrosequencing, indicating that the proposed method has great potential in genotyping of personalized medicine-related single nucleotide polymorphism.
- Aldehyde dehydrogenase 2,
- Gold nanoparticle probe,
- Visualized detection,
- Single nucleotide polymorphism,
- Invasive reaction
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