Smart Amp is a rapid, high-accuracy PCR diagnostic technology that helps both to identify a foreign agent in any biological material and to determine the mutations of that agent and its sensitivity to treatment.
How does Smart Amp works:
Smart Amp enables DNA amplification under the isothermal conditions using asymmetric primer set and Aac DNA polymerase, which exhibits a strand displacement reaction activity. It requires no DNA purification, and displays the high sensitivity of single-nucleotide precision, enabling identification of single nucleotide polymorphisms (SNPs), mutations, and other DNA targets.
The Smart Amplification process (Smart Amp) method uses isothermal nucleic acid amplification technology and employs a single reaction temperature (60-67ºC), which translates into simple and less expensive instrumentation. Smart Amp method facilitates not only highly sensitive detection of target DNA such as infectious diseases, but also discrimination of genetic polymorphisms or mutations in target.
The entire DNA amplification process in the Smart Amp method requires four primers: the turnback primer (TP), boost primer (BP), forward primer (FP), and outer primers (OP) . Comparing with PCR, Smart Amp produces continuous long double-stranded DNA concatenated target sequences as the amplified products by self-annealing and priming of 3’-terminal end of products based on TP and FP sequences. Additionally, DNA polymerase used Smart Amp has strand-displacement activity, and the polymerase synthesizes newly DNA strand with unthreading of double-stranded DNA. Therefore in Smart Amp reaction, each primer easily anneals to target DNA without denaturing of DNA by high-temperature, and DNA synthesis continuously occurs in isothermal condition. For precise design of primer sets for the reaction, Primers are selected based on those algorithms that consider free energy, probability of base-pairing, product size range, optimal melting temperature, and product size range.
Since Smart Amp method is carried out at 60-67ºC, it is able to combine a reverse-transcipase (RT)-reaction and Smart Amp-based DNA amplification reaction in one step, and the method is also available for genomic RNA detection. As example using this one-step reaction for clinical needs, we constructed highly sensitive detection kit of influenza A/B virus. The result of comparing with conventional method such as immunochromatography, our established RT-Smart Amp kit achieved 100 times higher sensitivity