Gold nanostructures for the multiplex detection of glucose-6-phosphate dehydrogenase gene mutations.
We describe a gold nanoparticle-based technique for the detection of single-base mutations in the glucose-6-phosphate dehydrogenase (G6PD) gene, a condition that can lead to neonatal jaundice and hemolytic anemia. The aim of this technique is to clearly distinguish different mutations frequently described within the Asian population from their wild-type counterparts and across different mutant variants. Gold nanoparticles of different sizes were synthesized, and each was conjugated with a single-strand DNA (ssDNA) sequence specific for a particular mutation in the G6PD gene. It was found that only mutant targets presented a characteristic band on the agarose gel, indicating the successful formation of dimeric nanostructures. No such dimer bands were observed for the wild-type targets. The difference in the relative dimer band levels allowed different mutant variants to be distinguished from one another. The technique was further validated using G6PD-deficient patient samples. This simple mutation detection method with direct result readout is amenable for rapid and mass screening of samples.
Department of Chemical and Biomolecular Engineering, Faculty of Engineering, National University of Singapore, Singapore 119260, Singapore.
Nianjia
N
Seow
Department of Pediatrics, National University Health System, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 119228, Singapore. Electronic address: paelaips@nus.edu.sg.
Poh San
PS
Lai
Department of Chemical and Biomolecular Engineering, Faculty of Engineering, National University of Singapore, Singapore 119260, Singapore. Electronic address: cheyly@nus.edu.sg.
Lin-Yue Lanry
LY
Yung
D004247:DNA / Q000032:analysis* / Q000378:metabolism; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000138:chemical synthesis / Q000378:metabolism; D019281:Dimerization; D004587:Electrophoresis, Agar Gel; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D010316:Particle Size; D017354:Point Mutation
D015342:DNA Probes; D004277:DNA, Single-Stranded; D006046:Gold; D004247:DNA; D005954:Glucosephosphate Dehydrogenase
Gel electrophoresis; Glucose-6-phosphate dehydrogenase (G6PD) deficiency; Gold nanoparticles; Multiplex detection; Mutation screening