tetano
Editor, Senior Moderator
Biosens Bioelectron. 2015 Apr 25;72:121-126. doi: 10.1016/j.bios.2015.04.078. [Epub ahead of print]
[h=1]Colorimetric monitoring of rolling circle amplification for detection of H5N1 influenza virus using metal indicator.[/h] Hamidi SV[SUP]1[/SUP], Ghourchian H[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] A new colorimetric method for monitoring of rolling circle amplification was developed. At first H[SUB]5[/SUB]N[SUB]1[/SUB] target hybrids with padlock probe (PLP) and then PLP is circularized upon the action of T[SUB]4[/SUB] ligase enzyme. Subsequently, the circular probe is served as a template for hyperbranched rolling circle amplification (HRCA) by utilizing Bst DNA polymerase enzyme. By improving the reaction, pyrophosphate is produced via DNA polymerization and chelates the Mg[SUP]2+[/SUP] in the buffer solution. This causes change in solution color in the presence of hydroxy naphthol blue (HNB) as a metal indicator. By using pH shock instead of heat shock and isothermal RCA reaction not only the procedure becomes easier, but also application of HNB for colorimetric detection of RCA reaction further simplifies the assay. The responses of the biosensor toward H[SUB]5[/SUB]N[SUB]1[/SUB] were linear in the concentration range from 0.16 to 1.20pM with a detection limit of 28 fM.
Copyright ? 2015 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] Bst DNA polymerase; H(5)N(1) virus; Hydroxy naphthol blue; Rolling circle amplification; pH shock
PMID: 25974174 [PubMed - as supplied by publisher]
[h=1]Colorimetric monitoring of rolling circle amplification for detection of H5N1 influenza virus using metal indicator.[/h] Hamidi SV[SUP]1[/SUP], Ghourchian H[SUP]2[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] A new colorimetric method for monitoring of rolling circle amplification was developed. At first H[SUB]5[/SUB]N[SUB]1[/SUB] target hybrids with padlock probe (PLP) and then PLP is circularized upon the action of T[SUB]4[/SUB] ligase enzyme. Subsequently, the circular probe is served as a template for hyperbranched rolling circle amplification (HRCA) by utilizing Bst DNA polymerase enzyme. By improving the reaction, pyrophosphate is produced via DNA polymerization and chelates the Mg[SUP]2+[/SUP] in the buffer solution. This causes change in solution color in the presence of hydroxy naphthol blue (HNB) as a metal indicator. By using pH shock instead of heat shock and isothermal RCA reaction not only the procedure becomes easier, but also application of HNB for colorimetric detection of RCA reaction further simplifies the assay. The responses of the biosensor toward H[SUB]5[/SUB]N[SUB]1[/SUB] were linear in the concentration range from 0.16 to 1.20pM with a detection limit of 28 fM.
Copyright ? 2015 Elsevier B.V. All rights reserved.
[h=4]KEYWORDS:[/h] Bst DNA polymerase; H(5)N(1) virus; Hydroxy naphthol blue; Rolling circle amplification; pH shock
PMID: 25974174 [PubMed - as supplied by publisher]