Open AccessAn efficient isothermal PCR method for on-site detection of nucleic acid
Author(s) -
Shiyin Zhang,
Jin Wang,
Zhihao Zhuo,
Xiaosong Su,
Mengyuan Chen,
Wendi Chen,
Tingdong Li,
Dongxu Zhang,
Xiaoping Min,
Shengxiang Ge,
Ningshao Xia
Publication year - 2019
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2018-0190
Subject(s) - loop mediated isothermal amplification , nucleic acid , isothermal process , multiplex polymerase chain reaction , convection , polymerase chain reaction , polycarbonate , biology , materials science , biological system , chemistry , chromatography , analytical chemistry (journal) , dna , biochemistry , gene , thermodynamics , composite material , physics
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.