Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay.
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.
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China.
Peihu
P
Fan
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China.
Xiaojun
X
Li
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China.
Weiheng
W
Su
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China; Key Laboratory for Molecular Enzymology & Engineering, Jilin University, Changchun, Jilin, China.
Wei
W
Kong
State Key Laboratory of Luminescence and Applications, Changchun Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun, Jilin, China.
Xianggui
X
Kong
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin, China.
Zhenxin
Z
Wang
Division of Human Immunodeficiency Virus/Acquired Immune Deficiency Syndrome and Sex-transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
Youchun
Y
Wang
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China; Key Laboratory for Molecular Enzymology & Engineering, Jilin University, Changchun, Jilin, China.
Chunlai
C
Jiang
School of Life Sciences, Jilin University, Changchun, Jilin, China; National Engineering Laboratory for Acquired Immune Deficiency Syndrome Vaccine, Jilin University, Changchun, Jilin, China.
Feng
F
Gao
D005456:Fluorescent Dyes / Q000737:chemistry; D015200:Fluoroimmunoassay / Q000379:methods*; D015483:HIV Antibodies / Q000276:immunology; D016655:HIV Core Protein p24 / Q000032:analysis* / Q000276:immunology; D015658:HIV Infections / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D006801:Humans; D058726:Nanoconjugates / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry* / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism*; D012680:Sensitivity and Specificity
D005456:Fluorescent Dyes; D015483:HIV Antibodies; D016655:HIV Core Protein p24; D058726:Nanoconjugates; D009841:Oligonucleotides; D012260:Ribonucleases