Jump to main content
Jump to site search

Issue 5, 2015
Previous Article Next Article

Real-time detection of H5N1 influenza virus through hyperbranched rolling circle amplification

Author affiliations

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.

Graphical abstract: Real-time detection of H5N1 influenza virus through hyperbranched rolling circle amplification

Back to tab navigation

Publication details

The article was received on 26 Oct 2014, accepted on 12 Jan 2015 and first published on 12 Jan 2015


Article type: Paper
DOI: 10.1039/C4AN01954G
Author version available: Download Author version (PDF)
Citation: Analyst, 2015,140, 1502-1509
  •   Request permissions

    Real-time detection of H5N1 influenza virus through hyperbranched rolling circle amplification

    S. V. Hamidi, H. Ghourchian and G. Tavoosidana, Analyst, 2015, 140, 1502
    DOI: 10.1039/C4AN01954G

Search articles by author

Spotlight

Advertisements