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

BioTechniques

Jin Wang¹  Dongxu Zhang¹  Shengxiang Ge¹  Xiaosong Su¹  Wendi Chen¹  Zhihao Zhuo²  Ningshao Xia²  Xiaoping Min²  Mengyuan Chen²  Shiyin Zhang²  Tingdong Li² 

[1] Molecular Diagnostics, Xiamen University, Xiamen, 361102, China;;1State Key Laboratory of Molecular Vaccinology &

关键词: convective PCR;    isothermal PCR;    molecular diagnostics;    point-of-care-test;    Rayleigh–Bénard convection;   

DOI  :  10.2144/btn-2018-0190

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown