Background: Currently available molecular method to detect HER2I655V polymorphism such as PCR-RFLP is
hampered by costly experimental method and post-PCR treatment requirment that make this technique is not meeting
for high-throughput analysis purpose. In this study we developed a accurate, simple, low cost and rapid test to detect
polymorphism at HER2 gene using SBR Green I based-melting curve method. Methods: Two forward allele-specific
primers and one common reverse primer were used then these primers were tested to discriminate known genotypes of
genomic templates (GG type or AA type) and genomic samples retrieved from breast cancer patients. Results: Melting
curve analysis derived from SYBR Green I-based allele-specific PCR with defined primers concentration and anneling
temperature at 54.3 °C showed good discrimination level of Tm peaks in which GG genotype melted at 89 °C slightly
higher than AA genotype which melted at 86 °C, while AG genotype harbored both of homozygous Tm characteristics.
Conclusions: This preliminary result will be as basic for further large-scale typing of HER2I655V polymorphism.
