tetano
Editor, Senior Moderator
Anal Chem. 2015 Apr 16. [Epub ahead of print]
[h=1]Recognition of dual targets by molecular beacon-based sensor: subtyping of influenza A virus.[/h] Lee CC, Liao YC, Lai YH, Chuang MC.
[h=3]Abstract[/h] A molecular beacon (MB)-based sensor to offer a decisive answer in combination with information originated from dual target inputs is designed. The system harnesses an assistant strand and thermodynamically favoured designation of unpaired nucleotides (UNs) to process the binary targets in 'AND gate' format and report fluorescence in 'off-on' mechanism via a formation of DNA four-way junction (4WJ). By manipulating composition of the UNs, the dynamic fluorescence difference between the binary targets-coexisting circumstance and any other scenario was maximized. Characteristic equilibrium constant (K), change of entropy (S), and association rate constant (k) between the association ('on') and dissociation ('off') states of the 4WJ were evaluated to understand unfolding behavior of MB in connection to its sensing capability. Favorable MB and UNs were furthermore designed towards analysis of genuine genetic sequences of hemagglutinin (HA) and neuraminidase (NA) in an influenza A H5N2 isolate. The MB-based sensor was demonstrated to yield a linear calibration range from 1.2 to 240 nM and detection limit of 120 pM. Furthermore, high-fidelity subtyping of influenza virus was implemented in sample of unpurified amplicons. The strategy opens an alternative avenue of MB-based sensors for dual targets towards applications in clinical diagnosis.
PMID: 25879394 [PubMed - as supplied by publisher]
[h=1]Recognition of dual targets by molecular beacon-based sensor: subtyping of influenza A virus.[/h] Lee CC, Liao YC, Lai YH, Chuang MC.
[h=3]Abstract[/h] A molecular beacon (MB)-based sensor to offer a decisive answer in combination with information originated from dual target inputs is designed. The system harnesses an assistant strand and thermodynamically favoured designation of unpaired nucleotides (UNs) to process the binary targets in 'AND gate' format and report fluorescence in 'off-on' mechanism via a formation of DNA four-way junction (4WJ). By manipulating composition of the UNs, the dynamic fluorescence difference between the binary targets-coexisting circumstance and any other scenario was maximized. Characteristic equilibrium constant (K), change of entropy (S), and association rate constant (k) between the association ('on') and dissociation ('off') states of the 4WJ were evaluated to understand unfolding behavior of MB in connection to its sensing capability. Favorable MB and UNs were furthermore designed towards analysis of genuine genetic sequences of hemagglutinin (HA) and neuraminidase (NA) in an influenza A H5N2 isolate. The MB-based sensor was demonstrated to yield a linear calibration range from 1.2 to 240 nM and detection limit of 120 pM. Furthermore, high-fidelity subtyping of influenza virus was implemented in sample of unpurified amplicons. The strategy opens an alternative avenue of MB-based sensors for dual targets towards applications in clinical diagnosis.
PMID: 25879394 [PubMed - as supplied by publisher]