Visual detection of cucumber green mottle mosaic virus based on terminal deoxynucleotidyl transferase coupled with DNAzymes amplification

Wang, Ying, Liu, Jing and Zhou, Hong 2019, Visual detection of cucumber green mottle mosaic virus based on terminal deoxynucleotidyl transferase coupled with DNAzymes amplification, Sensors, vol. 19, no. 6, doi: 10.3390/s19061298.

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Title Visual detection of cucumber green mottle mosaic virus based on terminal deoxynucleotidyl transferase coupled with DNAzymes amplification
Author(s) Wang, Ying
Liu, Jing
Zhou, Hong
Journal name Sensors
Volume number 19
Issue number 6
Article ID 1298
Total pages 12
Publisher MDPI
Place of publication Basel, Switzerland
Publication date 2019-03-14
ISSN 1424-8220
Keyword(s) DNAzyme
cascade amplification
cucumber green mottle mosaic virus
terminal deoxynucleotidyl transferase
visual detection
Summary A simple, rapid, and sensitive visual detection method for observing cucumber green mottle mosaic virus was reported based on the template-independent polymerization activity of terminal deoxynucleotidyl transferase (TdT), coupled with the cascade amplification of Mg2+-dependent DNAzyme and hemin/G-quadruplex DNAzyme. Briefly, the hybridized dsDNA of T1/P1 was cut into two parts at its position of 5'-AA↓CG↑TT-3' by the restricted enzyme AcII. The longer, newborn fragment originating from P1 was tailed at its 3'-end by oligo dG, and an intact enzymatic sequence of Mg2+-dependent DNAzyme was generated. The substrate sequence in the loop segment of the hairpin probe (HP) hybridized with the newborn enzymatic sequence and was cleaved into two parts in the presence of Mg2+. The locked G-quadruplex sequence in the stem segment of the HP was released, which catalyzed the oxidation of ABTS2- in the presence of H₂O₂, and the resulting solution turned green. A correlation between the absorbance and concentration of T1 was obtained in a range from 0.1 pM to 2 nM, with a detection limit of 0.1 pM. In addition to promoting a lower detection limit and shorter monitoring time, this method also demonstrated an excellent selectivity to single or double nucleotide changes. Therefore, the designed strategy provided a rapid and efficient platform for viral inspection and plant protection.
Language eng
DOI 10.3390/s19061298
Field of Research 0301 Analytical Chemistry
0906 Electrical and Electronic Engineering
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2019, The Authors
Persistent URL http://hdl.handle.net/10536/DRO/DU:30120756

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