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

Fan, Peihu; Li, Xiaojun; Su, Weiheng; Kong, Wei; Kong, Xianggui; Wang, Youchun; Jiang, Chunlai; Gao, Feng

doi:10.1371/journal.pone.0125701

Cited by 14 publications

(14 citation statements)

References 51 publications

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“…However, a low number of antigens will only accumulate a low number of detection antibodies, which will produce a correspondingly small signal and this will therefore affect the sensitivity of the immunoassays. This physical limitation can cause antigens to become undetectable in the window period of diagnostic tests 2 , 4 . For example, p24 antigen, a biomarker for early HIV infection, is only detectable by ELISA 14 days after the initial infection 4 .…”

Section: Introductionmentioning

confidence: 99%

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Hao

Chen

et al. 2017

Sci Rep

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The sensitivities of solid-phase immunoassays are limited by the quantity of detection antibodies bound to their antigens on the solid phase. Here, we developed a poly-protein G-expressing bacterium as an antibody-trapping microparticle to enhance the signals of immunoassays by increasing the accumulation of detection antibodies on the given antigen. Eight tandemly repeated fragment crystallisable (Fc) binding domains of protein G were stably expressed on the surface of Escherichia coli BL21 cells (termed BL21/8G). BL21/8G cells showed a higher avidity for trapping antibodies on their surface than monomeric protein G-expressing BL21 (BL21/1G) cells did. In the sandwich enzyme-linked immunosorbent assay (ELISA), simply mixing the detection antibody with BL21/8G provided a detection limit of 6 pg/mL for human interferon-α (IFN-α) and a limit of 30 pg/mL for polyethylene glycol (PEG)-conjugated IFN-α (Pegasys), which are better than that of the traditional ELISA (30 pg/mL for IFN-α and 100 pg/mL for Pegasys). Moreover, the sensitivity of the Western blot for low-abundance Pegasys (0.4 ng/well) was increased by 25 folds upon mixing of an anti-PEG antibody with BL21/8G cells. By simply being mixed with a detection antibody, the poly-protein G-expressing bacteria can provide a new method to sensitively detect low-abundance target molecules in solid-phase immunoassays.

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Section: Introductionmentioning

confidence: 99%

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Hao

Chen

et al. 2017

Sci Rep

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“…[132] In addition, this approach reduces the cost by 20-fold in comparison to RT-PCR. In this assay, named NLFOA (nuclease-linked fluorescence oligonucleotide assay), nanoparticles coupled with several streptavidin molecules interacts specifically with the biotinylated TurboNuclease for detection of the HIV-1 p24 antigen.…”

Section: Applications Of Nanoscale Avidin Systems In Diagnosis Andmentioning

confidence: 99%

“…The NLFOA provides a highly sensitive assay system for the clinical diagnosis of HIV or other infectious diseases. [132]…”

Section: Applications Of Nanoscale Avidin Systems In Diagnosis Andmentioning

confidence: 99%

The principles and applications of avidin-based nanoparticles in drug delivery and diagnosis

Jain

Cheng

2017

Journal of Controlled Release

220

286

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Avidin-biotin interaction is one of the strongest non-covalent interactions in the nature. Avidin and its analogues have therefore been extensively utilized as probes and affinity matrices for a wide variety of applications in biochemical assays, diagnosis, affinity purification, and drug delivery. Recently, there has been a growing interest in exploring this non-covalent interaction in nanoscale drug delivery systems for pharmaceutical agents, including small molecules, proteins, vaccines, monoclonal antibodies, and nucleic acids. Particularly, the ease of fabrication without losing the chemical and biological properties of the coupled moieties makes the avidin-biotin system a versatile platform for nanotechnology. In addition, avidin-based nanoparticles have been investigated as diagnosis systems for various tumors and surface antigens. In this review, we will highlight the various fabrication principles and biomedical applications of avidin-based nanoparticles in drug delivery and diagnosis. The structures and biochemical properties of avidin, biotin and their respective analogues will also be discussed.

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“…RSD of 2.36% reveals the high precision of the measurement by this biosensor. According to the literature, most of the HIV biosensors are based on p24 antigen immunoassay [46,47] or HIV protease [48] measurement or enzyme and protein biosensor modifications which require special care and conditions [49]. A recent research has been made on HIV RNA target measurement which also suffer from RNA decomposition [50].…”

Section: The Analytical Performances Of the Biosensor For Hiv1 Detectionmentioning

confidence: 99%

Closed Bipolar Electrochemistry for the Detection of Human Immunodeficiency Virus Short Oligonucleotide

Poorghasem

Saberi

Shayan

et al. 2016

Electrochimica Acta

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Although a monumental progress has been made in case of Human Immunodeficiency Virus researches, there are still more than 34 million people living with this viral disease. Therefore, the development of a fast and cost-effective protocol that can sensitively and selectively detect HIV gene sequence is essential. In the present study, a protocol for the molecular detection of HIV genome taking advantage of closed bipolar electrochemistry integrated with electrogenerated chemiluminescence detection is described. In this case, a closed split bipolar electrochemical cell is applied to detect the short specific oligonucleotide target of HIV1. Anthraquinone disulfonic acid, as a negative electroactive reporter, intercalates with hybridized double-strand DNA on the sensing site. By reduction of redox reporter (AQDS) on the cathodic pole, luminol is oxidized on the anodic pole and subsequent oxidized diazo product generates light that is followed using a photomultiplier tube. The experimental condition is optimized to obtain the highest sensitivity. The results demonstrate the high sensitivity and selectivity of this fast and economical method toward Human Immunodeficiency Virus genome detection in the range of 0.1 to 300 nM with the detection limit of 30 pM (S/N = 3).

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

Cited by 14 publications

References 51 publications

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

The principles and applications of avidin-based nanoparticles in drug delivery and diagnosis

Closed Bipolar Electrochemistry for the Detection of Human Immunodeficiency Virus Short Oligonucleotide

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