Colorimetric monitoring of rolling circle amplification for detection of H5N1 influenza virus using metal indicator
作者： Seyed Vahid Hamidi , Hedayatollah Ghourchian
DOI: 10.1016/J.BIOS.2015.04.078
关键词:
摘要: Abstract A new colorimetric method for monitoring of rolling circle amplification was developed. At first H 5 N 1 target hybrids with padlock probe (PLP) and then PLP is circularized upon the action T 4 ligase enzyme. Subsequently, circular served as a template hyperbranched (HRCA) by utilizing Bst DNA polymerase By improving reaction, pyrophosphate produced via polymerization chelates Mg 2+ in buffer solution. This causes change solution color presence hydroxy naphthol blue (HNB) metal indicator. using pH shock instead heat isothermal RCA reaction not only procedure becomes easier, but also application HNB detection further simplifies assay. The responses biosensor toward were linear concentration range from 0.16 to 1.20 pM limit 28 fM.
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