Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification

Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification Publisher Pubmed

Hamidi SV¹ ; Ghourchian H^{1, 2} ; Tavoosidana G³

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1. Laboratory of Microanalysis, Institute of Biochemistry and Biophysics, University of Tehran, P.O. Box 131451384, Tehran, Iran
2. Nanobiomedicine Center of Excellence, Nanoscience and Nanotechnology Research Center, University of Tehran, P.O. Box 131451384, Tehran, Iran
3. Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, P.O. Box 1417755469, Tehran, Iran

Source: Analyst Published:2015

Abstract

An isothermal amplification method was developed for the sensitive detection of the H5N1 influenza virus. The padlock probe specifically bound to the H5N1 target and circularized with T4 DNA ligase enzyme. Then this circular probe was amplified by hyperbranched rolling circle amplification (HRCA) using Phi29 DNA polymerase. The fluorescence intensity was recorded at different intervals by intercalation of SYBR green molecules into the double-stranded product of the HRCA reaction. At an optimum time of 88 min, a calibration plot with fine linearity was obtained. Using HRCA based on a padlock probe and Phi29 DNA polymerase, high selectivity and sensitivity were achieved. The biosensor response was linear toward H5N1 in the concentration range from 10 fM to 0.25 pM, with a detection limit of 9 fM at a signal/noise ratio of 3. By replacing the heat shock with pH shock, not only was the procedure for detection of H5N1 influenza simplified, but also the DNA molecules were protected from possible breaking at high temperature. This journal is © The Royal Society of Chemistry.

Style	Citing Format
MLA	Hamidi SV, Ghourchian H, Tavoosidana G. "Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification." Analyst, vol. 140, no. 5, 2015, pp. 1502-1509.
APA	Hamidi SV, Ghourchian H, Tavoosidana G (2015). Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification. Analyst, 140(5), 1502-1509.
Chicago	Hamidi SV, Ghourchian H, Tavoosidana G. "Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification." Analyst 140, no. 5 (2015): 1502-1509.
Harvard	Hamidi SV, Ghourchian H, Tavoosidana G (2015) 'Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification', Analyst, 140(5), pp. 1502-1509.
Vancouver	Hamidi SV, Ghourchian H, Tavoosidana G. Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification. Analyst. 2015;140(5):1502-1509.
BibTex	@article{ author = {Hamidi SV and Ghourchian H and Tavoosidana G}, title = {Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification}, journal = {Analyst}, volume = {140}, number = {5}, pages = {1502-1509}, year = {2015} }
RIS	TY - JOUR AU - Hamidi SV AU - Ghourchian H AU - Tavoosidana G TI - Real-Time Detection of H5n1 Influenza Virus Through Hyperbranched Rolling Circle Amplification JO - Analyst VL - 140 IS - 5 SP - 1502 EP - 1509 PY - 2015 ER -

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