Strand-Exchange Nucleic Acid Circuitry with Enhanced Thermo-and Structure- Buffering Abilities Turns Gene Diagnostics Ultra-Reliable and Environmental Compatible

• Published in: Scientific reports Vol. 6; no. 1; p. 36605
• Main Authors: Zhu, Zhentong, Tang, Yidan, Jiang, Yu Sherry, Bhadra, Sanchita, Du, Yan, Ellington, Andrew D., Li, Bingling
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 04.11.2016; Nature Publishing Group
• Subjects: 38/22; 38/32; 38/71; 38/77; 631/45/147; 692/700/139/1512; 96/10; 96/33; 96/34; 96/95; Acids; Circuits; High temperature; Humanities and Social Sciences; Hybridization; Middle East respiratory syndrome; Molecular Diagnostic Techniques - methods; Molecular Diagnostic Techniques - standards; multidisciplinary; Nucleic Acid Conformation; Nucleic acids; Nucleic Acids - chemistry; Real-Time Polymerase Chain Reaction; Reproducibility of Results; Science; Temperature; Temperature effects; Temperature requirements
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep36605

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 −19 M 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.