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Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay.

Publication ,  Journal Article
Fan, P; Li, X; Su, W; Kong, W; Kong, X; Wang, Z; Wang, Y; Jiang, C; Gao, F
Published in: PLoS One
2015

The relatively high detection limit of the Enzyme-linked immunosorbent assay (ELISA) prevents its application for detection of low concentrations of antigens. To increase the sensitivity for detection of HIV-1 p24 antigen, we developed a highly sensitive nuclease-linked fluorescence oligonucleotide assay (NLFOA). Two major improvements were incorporated in NLFOA to amplify antibody-antigen interaction signals and reduce the signal/noise ratio; a large number of nuclease molecules coupled to the gold nanoparticle/streptavidin complex and fluorescent signals generated from fluorescent-labeled oligonucleotides by the nuclease. The detection limit of p24 by NLFOA was 1 pg/mL, which was 10-fold more sensitive than the conventional ELISA (10 pg/mL). The specificity was 100% and the coefficient of variation (CV) was 7.8% at low p24 concentration (1.5 pg/mL) with various concentrations of spiked p24 in HIV-1 negative sera. Thus, NLFOA is highly sensitive, specific, reproducible and user-friendly. The more sensitive detection of low p24 concentrations in HIV-1-infected individuals by NLFOA could allow detection of HIV-1 infections that are missed by the conventional ELISA at the window period during acute infection to further reduce the risk for HIV-1 infection due to the undetected HIV-1 in the blood products. Moreover, NLFOA can be easily applied to more sensitive detection of other antigens.

Duke Scholars

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Published In

PLoS One

DOI

EISSN

1932-6203

Publication Date

2015

Volume

10

Issue

4

Start / End Page

e0125701

Location

United States

Related Subject Headings

  • Sensitivity and Specificity
  • Ribonucleases
  • Oligonucleotides
  • Nanoconjugates
  • Humans
  • HIV Infections
  • HIV Core Protein p24
  • HIV Antibodies
  • General Science & Technology
  • Fluoroimmunoassay
 

Citation

APA
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MLA
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Fan, P., Li, X., Su, W., Kong, W., Kong, X., Wang, Z., … Gao, F. (2015). Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay. PLoS One, 10(4), e0125701. https://doi.org/10.1371/journal.pone.0125701
Fan, Peihu, Xiaojun Li, Weiheng Su, Wei Kong, Xianggui Kong, Zhenxin Wang, Youchun Wang, Chunlai Jiang, and Feng Gao. “Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay.PLoS One 10, no. 4 (2015): e0125701. https://doi.org/10.1371/journal.pone.0125701.
Fan P, Li X, Su W, Kong W, Kong X, Wang Z, et al. Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay. PLoS One. 2015;10(4):e0125701.
Fan, Peihu, et al. “Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay.PLoS One, vol. 10, no. 4, 2015, p. e0125701. Pubmed, doi:10.1371/journal.pone.0125701.
Fan P, Li X, Su W, Kong W, Kong X, Wang Z, Wang Y, Jiang C, Gao F. Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay. PLoS One. 2015;10(4):e0125701.

Published In

PLoS One

DOI

EISSN

1932-6203

Publication Date

2015

Volume

10

Issue

4

Start / End Page

e0125701

Location

United States

Related Subject Headings

  • Sensitivity and Specificity
  • Ribonucleases
  • Oligonucleotides
  • Nanoconjugates
  • Humans
  • HIV Infections
  • HIV Core Protein p24
  • HIV Antibodies
  • General Science & Technology
  • Fluoroimmunoassay