Allele specific polymerase chain reaction for detection of the genetic polymorphism of NMDA receptor

Allele specific polymerase chain reaction method (AS-PCR) was developed, validated and employed to determine the type and frequency of genetic polymorphism in NMDA receptor gene, GRIN2B for single nucleotide polymorphism rs10845840. The study has been approved by the relevant human ethics committee. Blood samples were collected in sodium-citrate tubes and stored at −80°C until further analysis. The genomic DNA was extracted from the whole blood using the salting out technique. Allele specific primers were designed using the appropriate software. The polymerase chain reaction method was optimised and validated using DNA sequencing technique. Sixty samples from healthy volunteers were analysed. The results demonstrated that 85% of the subjects had CC allele, 12% had CT allele and 3% had TT allele. The assay would be useful for detection of the polymorphism in GRIN2B gene since the variations that occurred in the analysed samples were more than 1%. Recently, the gene has been reported to be associated with human brain volume (temporal lobe). It was also reported that the reduced temporal lobe volume in the elderly was associated with an increased in Alzheimer´s disease risk. The optimised method will be used subsequently to elucidate the type and frequency of GRIN2B polymorphism among Alzheimer´s disease patients in Malaysia.