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An efficient isothermal PCR method for on-site detection of nucleic acid

Author:
Zhang, Shiyin  Wang, Jin  Zhuo, Zhihao  Su, Xiaosong  Chen, Mengyuan  Chen, Wendi  Li, Tingdong  Zhang, Dongxu  Min, Xiaoping  Ge, Shengxiang  Xia, Ningshao  


Journal:
BIOTECHNIQUES


Issue Date:
2019


Abstract(summary):

Convective PCR (CPCR) is an isothermal nucleic acid amplification technology; however, natural convection exhibits a chaotic and multiplex flow state, resulting in low amplification efficiency and specificity. We placed a polycarbonate strip (p-strip) inside reaction tubes to induce circumfluence by blocking the inner ring that originally allowed fluid to flow at suboptimal temperatures. Moreover, we constructed a dual-temperature instrument to provide appropriate denaturing and annealing zones for CPCR. Tubes containing p-strips exhibited significantly improved efficiency, sensitivity and specificity. For real-time detection, the variation coefficients of three replicates having the same concentrations were less than 2% in more than half of the cases, indicating improved CPCR amplification and potential as a commercial on-site nucleic acid diagnosis tool.


Page:
63---69


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