Double-Antigen Lateral Flow Immunoassay for the Detection of Anti-HIV-1 and -2 Antibodies Using Upconverting Nanoparticle Reporters

Abstract

Rapid diagnostic tests (RDTs) are often used for the detection of anti-human immunodeficiency virus (HIV) antibodies in remote locations in low- and middle-income countries (LMIC) with low or limited access to central laboratories. The typical format of an RDT is a lateral flow assay (LFA) with visual interpretation prone to subjectivity. This risk of misinterpretation can be overcome with luminescent upconverting nanoparticle reporters (UCNPs) measured with a miniaturized easy-to-use reader instrument. An LFA with UCNPs for anti-HIV-1/2 antibodies was developed and the assay performance was evaluated extensively with challenging patient sample panels. Sensitivity (n = 145) of the UCNP-LFA was 96.6% (95% CI: 92.1–98.8%) and specificity (n = 309) was 98.7% (95% CI: 96.7–99.7%). Another set of samples (n = 200) was used for a comparison between the UCNP-LFA and a conventional visual RDT. In this comparison, the sensitivities for HIV-1 were 96.4% (95% CI: 89.8–99.3%) and 97.6% (95% CI: 91.6–99.7%), for the UCNP-LFA and conventional RDT, respectively. The specificity was 100% (95% CI: 96.4–100%) for both assays. The developed UCNP-LFA demonstrates the applicability of UCNPs for the detection of anti-HIV antibodies. The signal measurement is done by a reader instrument, which may facilitate automated result interpretation, archiving and transfer of data from de-centralized locations.

Toward Next Generation Lateral Flow Assays: Integration of Nanomaterials

“…Integrated filters are also commonly used in LFAs that analyze whole blood samples, primarily for the removal of red blood cells. Two commercially available plasma separation membranes are Cytosep and Vivid plasma; , examples for their use in LFAs include the detection of the capsid protein VP72 from the African swine fever virus and the diagnosis of SARS-CoV-2 and antihuman immunodeficiency virus (HIV) …”

“…Two commercially available plasma separation membranes are Cytosep and Vivid plasma; 161,166 examples for their use in LFAs include the detection of the capsid protein VP72 from the African swine fever virus 167 and the diagnosis of SARS-CoV-2 168 and antihuman immunodeficiency virus (HIV). 169 One of the drawbacks to these types of filters is that the small pore sizes of the membranes (<9 μm) can promote blood clotting (Figure 12Ai), which limits the plasma/serum separation yield to just 10−30%. 170 To this end, Lu et al have developed a low-cost (<$0.10) cross-flow filtration pad that increases the separation yield by up to 86%.…”

Lateral flow assays (LFA) as an alternative medical diagnosis method for detection of virus species: The intertwine of nanotechnology with sensing strategies

“…An Upcon reader device detected the emitted signals from excited UC NPs, and SAS JMP Pro 14 statistics software interpreted the signals to detect Abs of HIV-1 and HIV-2 quantitatively ( Figure 4 B). This device was evaluated and reached a sensitivity of 96.6% and a specificity of 98.7% [ 110 ].…”

Challenges in the Detection of SARS-CoV-2: Evolution of the Lateral Flow Immunoassay as a Valuable Tool for Viral Diagnosis

“…LFIA test detects the target molecule on an absorbent membrane with antibodies aligned to form the test and control lines; the signal is analyzed qualitatively in a visual or semi-quantitative reading [ 18 , 20 ] ( Figure 1 ). LFIA is a valuable tool that has been used for detecting, diagnosing, and monitoring various viruses such as human immunodeficiency virus (HIV), human adenovirus, influenza A H1N1 virus, and SARS-CoV-2 [ 21 , 22 , 23 ].…”