Condition Optimization Of SAMP And Study On Detection Of HBV

The nucleic acid signal amplification techniques are often used for the detection of trace nucleic acids. In this paper, the orthogonal test was used to optimize the reaction condition of Single primer-triggered isothermal amplification for double-stranded DNA Detection (SAMP), and then applied it to the practical HBV sample.In this paper, the amount of polymerase, nicking enzyme, the concentration of primer and temperature were optimized by orthogonal test, and got an optimal combinations, when the amount of polymerase enzyme is 0.05 ?L, nicking enzyme is 0.4 ?L, the concentration of primer is 5.0�10-7 M and the temperature is 55?, it could get higher sensitivity and lower detection background limit. 1.0�10-17 M target dsDNA can be detected in the best reaction condition.In this paper, the gold nanoparticles was also used to analyse the amplification results. After 35 min rection of SAMP, the gold nanoparticles was add to achieve colorimetric detection. Results showed that high concentrations of the target system had a distinct color change, but low concentrations, as well as the blank had not changed color. Therefore, a sensitive colorimetric detection of target DNA could be achieved by this method.Finally, HBV sample was detected by SAMP in the optimal conditions. In this paper, primers for the types of HBV-B, HBV-C and HBV-D were designed and detected by SAMP. The detection limit of HBV-D can be reached 1.0�10-17 M, and SAMP showed better regularity and higher fluorescence sensitivity than PCR. In addition, the primers for HBV-B type, HBV-C need to be further optimized to achieve better detection limits.