Upconverting nanoparticle reporter–based highly sensitive rapid lateral flow immunoassay for hepatitis B virus surface antigen

Abstract

Detection of hepatitis B Virus surface antigen (HBsAg) is an established method for diagnosing both acute and chronic hepatitis B virus (HBV) infection. In addition to enzyme immunoassays (EIAs), rapid diagnostic tests (RDTs) are available for the detection of HBsAg in resource-poor settings. However, the available RDTs have inadequate sensitivity and therefore are not suitable for diagnosis of patients with low levels of HBsAg and for blood screening. To provide a high-sensitivity RDT, we developed a lateral flow immunoassay (LFIA) for HBsAg utilizing upconverting nanoparticle (UCNP) reporter. The UCNP-LFIA can use whole blood, serum, or plasma and the results can be read in 30 min using a reader device. When compared with a commercial conventional visually read LFIA, the developed UCNP-LFIA had a Limit of Detection (LoD) of 0.1 IU HBsAg/ml in spiked serum, whereas the LoD of the conventional LFIA was 3.2 IU HBsAg/ml. The developed UCNP-LFIA fulfills the WHO criterion for blood screening (LoD ≤ 0.13 IU HBsAg/ml) in terms of LoD. The UCNP-LFIA and conventional LFIA were evaluated with well-characterized sample panels. The UCNP-LFIA detected 20/24 HBsAg-positive samples within the HBsAg Performance Panel and 8/10 samples within the Mixed Titer Performance Panel, whereas the conventional LFIA detected 8/24 and 4/10 samples in these panels, respectively. The performance of the assays was further evaluated with HBsAg-positive (n = 108) and HBsAg-negative (n = 315) patient samples. In comparison with a central laboratory test, UCNP-LFIA showed 95.4% (95% CI: 89.5–98.5%) sensitivity whereas sensitivity of the conventional LFIA was 87.7% (95%CI: 79.9–93.3%).

Latest Trends in Lateral Flow Immunoassay (LFIA) Detection Labels and Conjugation Process

“…Upconverting nanoparticles have been used as reporters on rapid lateral flow immunoassay in a few applications. Martiskainen et al (2021) [5] developed a rapid test for the diagnosis of hepatitis B virus surface antigen, using upconverting nanoparticles as reporter. The assay reported improved specifications than the conventional visually read LFIA, the limit of detection being 0.1 IU HBsAg/ml in spiked serum, compared to 3.2 IU HBsAg/ml.…”

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

“…This UC NP-LFA was evaluated and compared with a conventional LFA. UC NP-LFA reached a sensitivity of 95.4%, while the conventional LFA's sensitivity was 87.7% [ 62 ].…”

Lateral Flow Immunoassay Based on Time-Resolved Fluorescence Microspheres for Rapid and Quantitative Screening CA199 in Human Serum

“…It has been widely used because of its advantages of simplicity, rapidity, convenience, low cost, and high efficacy. The mature LFIA has become a commonly used POCT technique in clinical diagnosis of various diseases, such as early-stage cancer [ 14 ], sexually transmitted diseases [ 15 ], AIDS [ 16 ], COVID-19 [ 17 ], and so forth. Our group have successfully developed LFIAs to rapidly detect SARS-CoV-2 nucleocapsid protein [ 18 ], IgG/IgM [ 19 ] and neutralizing antibody [ 20 ].…”