tetano
Editor, Senior Moderator
Talanta. 2019 Jan 15;192:189-196. doi: 10.1016/j.talanta.2018.09.049. Epub 2018 Sep 15.
[h=1]Self-enzyme chemiluminescence immunoassay capable of rapidly diagnosing the infection of influenza A (H1N1) virus.[/h] Kyme H[SUP]1[/SUP], Lee CT[SUP]1[/SUP], Kim YT[SUP]2[/SUP], Lee JH[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] A highly sensitive self-enzyme immunoassay with 1,1'-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed for the first time to quantify influenza A (H1N1) virus. A specific capture antibody, capable of capturing hemagglutinin (HA) subtypes of H1N1, was immobilized on the surface of the magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposite. Neuraminidase (NA) subtype of H1N1 was acts as an enzyme in the self-enzyme immunoassay. A sample mixed with HA antibodies immobilized on the surface of magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposites was incubated for 1 h at 37 ?C. After the incubation, magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposites separated with a magnetic bar were washed 3 times using phosphate buffered saline with Tween-20 (PBST). Then, 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA), a fluorogenic substrate of NA, was added and incubated for 10 min to produce 4-Methylumbelliferone (4-MU) from the enzyme reaction between MUNANA and NA of H1N1. After the incubation, the solution containing 4-MU emitted bright light with the addition of ODI-CL reagents (e.g., H[SUB]2[/SUB]O[SUB]2[/SUB] and ODI). The relative CL intensity of 4-MU was proportionally enhanced with the increase of H1N1. Also, the brightness and color of 4-MU light emitted from the self-enzyme immunoassay was dependent on pH. The self-enzyme immunoassay was able to analyze trace levels of influenza A (H1N1) virus with good accuracy, precision, reproducibility and excellent selectivity. The limit of detection (LOD = 3σ/slope) was as low as 0.19 U/ml. We expect that the self-enzyme immunoassay can be applied as a cost-effective and rapid method capable of selectively quantifying a specific influenza A virus such as H1N1, H3N2, and H5N1.
[h=4]KEYWORDS:[/h] 1,1′-Oxalyldiimidazole (ODI); 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA); Chemiluminescence; Enzyme immunoassay; Influenza A virus; Nanocomposites; Neuraminidase
PMID: 30348377 DOI: 10.1016/j.talanta.2018.09.049
[h=1]Self-enzyme chemiluminescence immunoassay capable of rapidly diagnosing the infection of influenza A (H1N1) virus.[/h] Kyme H[SUP]1[/SUP], Lee CT[SUP]1[/SUP], Kim YT[SUP]2[/SUP], Lee JH[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] A highly sensitive self-enzyme immunoassay with 1,1'-oxalyldiimidazole chemiluminescence (ODI-CL) detection was developed for the first time to quantify influenza A (H1N1) virus. A specific capture antibody, capable of capturing hemagglutinin (HA) subtypes of H1N1, was immobilized on the surface of the magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposite. Neuraminidase (NA) subtype of H1N1 was acts as an enzyme in the self-enzyme immunoassay. A sample mixed with HA antibodies immobilized on the surface of magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposites was incubated for 1 h at 37 ?C. After the incubation, magnetic Au-Fe[SUB]3[/SUB]O[SUB]4[/SUB] nanocomposites separated with a magnetic bar were washed 3 times using phosphate buffered saline with Tween-20 (PBST). Then, 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA), a fluorogenic substrate of NA, was added and incubated for 10 min to produce 4-Methylumbelliferone (4-MU) from the enzyme reaction between MUNANA and NA of H1N1. After the incubation, the solution containing 4-MU emitted bright light with the addition of ODI-CL reagents (e.g., H[SUB]2[/SUB]O[SUB]2[/SUB] and ODI). The relative CL intensity of 4-MU was proportionally enhanced with the increase of H1N1. Also, the brightness and color of 4-MU light emitted from the self-enzyme immunoassay was dependent on pH. The self-enzyme immunoassay was able to analyze trace levels of influenza A (H1N1) virus with good accuracy, precision, reproducibility and excellent selectivity. The limit of detection (LOD = 3σ/slope) was as low as 0.19 U/ml. We expect that the self-enzyme immunoassay can be applied as a cost-effective and rapid method capable of selectively quantifying a specific influenza A virus such as H1N1, H3N2, and H5N1.
[h=4]KEYWORDS:[/h] 1,1′-Oxalyldiimidazole (ODI); 4-Methylumbelliferyl-N-acetyl-α-D-neuraminic acid (MUNANA); Chemiluminescence; Enzyme immunoassay; Influenza A virus; Nanocomposites; Neuraminidase
PMID: 30348377 DOI: 10.1016/j.talanta.2018.09.049