Strand-Exchange Nucleic Acid Circuitry with Enhanced Thermo-and Structure- Buffering Abilities Turns Gene Diagnostics Ultra-Reliable and Environmental Compatible.
Catalytic hairpin assembly (CHA) is one of the most promising nucleic acid amplification circuits based on toehold-mediated strand exchange reactions. But its performance is usually ruined by fluctuated environmental temperatures or unexpected self-structures existing in most real-world targets. Here we present an amide-assistant mechanism that successfully reduces the prevalence of these problems for CHA and maximizes its thermo- and structure- buffering abilities. Such an organic amide-promoted CHA (shortened as OHT-CHA) can unprecedentedly amplify through 4 °C to 60 °C without rebuilding sequences or concerning target complexity. We are then for the first time able to employ it as a direct and universal signal booster for loop mediated isothermal reaction (LAMP). LAMP is one of the most promising point-of-care (POC) gene amplifiers, but has been hard to detect precisely due to structured products and haunted off-target amplicons. OHT-CHA guarantees a significant and reliable signal for LAMP reaction amplified from as little as 10-19M virus gene. And one single set of OHT-CHA is qualified to any detection requirement, either in real-time at LAMP running temperature (~60 °C), or at end-point on a POC photon counter only holding environmental temperatures fluctuating between 4 °C to 42 °C.
State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun, Jilin, 130022, P.R. China.|University of Chinese Academy of Sciences, Beijing, 100039, China.
Zhentong
Z
Zhu
State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun, Jilin, 130022, P.R. China.|University of Chinese Academy of Sciences, Beijing, 100039, China.
Yidan
Y
Tang
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas 78712, United States.
Yu Sherry
YS
Jiang
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas 78712, United States.
Sanchita
S
Bhadra
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas 78712, United States.
Yan
Y
Du
Center for Systems and Synthetic Biology, University of Texas at Austin, Austin, Texas 78712, United States.
Andrew D
AD
Ellington
State Key Lab of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Science, Changchun, Jilin, 130022, P.R. China.
Bingling
B
Li
D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013696:Temperature
D009696:Nucleic Acids