摘要
Gold nanoparticles(AuNPs)have been reported to induce faster heat transfer of polymerase chain reaction(PCR)solution to enhance PCR efficiency.AuNPs can also increase the specificity of PCR by functioning analogously to single-stranded DNA binding protein(SSB).However,the simple structure of AuNPs makes it difficult to determine how AuNPs affect PCR efficiency and specificity.This study aimed to elucidate the effect of AuNPs on PCR efficiency by altering the denaturation times and annealing temperatures.In addition,comparative study of the effect of AuNPs on PCR inhibition caused by two competitive primers allowed investigation of the mechanisms of AuNP-enhanced PCR accuracy.Finally,heat-treated salmon sperm DNA was used to evaluate whether AuNPs could eliminate the inhibitory PCR effect caused by DNA impurities.This study demonstrated that enhanced thermal conductivity by AuNPs was the main mechanism for increased PCR efficiency and specificity.AuNPs promoted efficient double-stranded DNA template unwinding and dissociation between mismatched primers,DNA fragments and template,and enhanced PCR efficiency and specificity simultaneously.Thus,this significant finding suggests the future use of AuNP-assisted PCR in different fields,especially in rapid clinical diagnosis and screening by PCR.
Gold nanoparticles （AuNPs） have been reported to induce faster heat transfer of polymerase chain reaction （PCR） solution to enhance PCR efficiency. AuNPs can also increase the specificity of PCR by functioning analogously to single-stranded DNA bind- ing protein （SSB）. However, the simple structure of AuNPs makes it difficult to determine how AuNPs affect PCR efficiency and specificity. This study aimed to elucidate the effect of AuNPs on PCR efficiency by altering the denaturation times and annealing temperatures. In addition, comparative study of the effect of AuNPs on PCR inhibition caused by two competitive primers al- lowed investigation of the mechanisms of AuNP-enhanced PCR accuracy. Finally, heat-treated salmon sperm DNA was used to evaluate whether AuNPs could eliminate the inhibitory PCR effect caused by DNA impurities. This study demonstrated that en- hanced thermal conductivity by AuNPs was the main mechanism for increased PCR efficiency and specificity. AuNPs promoted efficient double-stranded DNA template unwinding and dissociation between mismatched primers, DNA fragments and template, and enhanced PCR efficiency and specificity simultaneously. Thus, this significant finding suggests the future use of AuNP-assisted PCR in different fields, especially in rapid clinical diagnosis and screening by PCR.
基金
supported by the National Natural Science Foundation of China(20907017)
the National Basic Research Program of China(2009CB421605)
the Program for New Century Excellent Talents in University(NCET-11-0964)
