Abstract
Hepatitis B is a globally prevalent viral infectious disease caused by the hepatitis B virus (HBV). In this study, an immunochromatographic assay (ICA) for the rapid detection of hepatitis B preS2 antigen (preS2Ag) was established. The magnetic nanoparticles (MNPs) indirectly labelled with goat anti-mouse (GAM) secondary antibody were applied as a nanoprobe for free preS2 antibody (preS2Ab) capturing and signal amplification. By employing sample pre-incubation processing as well, preS2Ag-preS2Ab was sufficiently caught by the GAM-MNPs probe in 5 min. A qualitative sensitivity of 625 ng/mL was obtained by naked-eye observation within 15–20 min. A standard curve (0–5000 ng/mL) was established, with a quantitative limit of detection (LOD) of 3.6 ng/mL, based on the stability and penetrability of the magnetic signal characteristics. The proposed method for preS2Ag was rapid (~25 min, cf. ELISA ~4 h) and had a good accuracy, which was verified using an ELISA kit (relative error < 15%). Large equipment and skilled technicians were not required. The sensitivity and specificity of the developed GAM-MNPs-ICA method were 93.3% and 90% in clinical serum samples (n = 25), respectively. A good detection consistency (84%) was observed between the developed ICA method and 2 types of commercial ELISA kits, indicating that the GAM-MNPs-ICA has a potential application in large-scale screening for and point-of-care diagnosis of hepatitis B or other infectious diseases.
Highlights
Hepatitis B remains a great threat to public health
It is estimated that more than 257 million people have suffered from chronic hepatitis B infection worldwide, and almost 9 million people have died from liver failure, cirrhosis, hepatocellular carcinoma, and other related diseases caused by hepatitis B virus (HBV) [1]
In order to ensure a sufficient combination of antigen and antibody and capture a trace amount of order to ensure a sufficient combination of antigen and antibody and capture a trace amount of preS2 antigen (preS2Ag), a sample pre-incubation of preS2Ag, preS2 antibody (preS2Ab), and goat anti-mouse (GAM)-magnetic nanoparticles (MNPs) probes was applied to preS2Ag, a sample pre-incubation of preS2Ag, preS2Ab, and GAM-MNPs probes was applied form a complex of GAM-MNPs-preS2Ab-preS2Ag (Figure 1b)
Summary
Hepatitis B remains a great threat to public health. It is estimated that more than 257 million people have suffered from chronic hepatitis B infection worldwide, and almost 9 million people have died from liver failure, cirrhosis, hepatocellular carcinoma, and other related diseases caused by HBV [1].Hepatitis B surface antigen (HBsAg) is the envelope protein of HBV, which is the most conventional serological marker for the screening, diagnosis, and antiviral therapy evaluation of hepatitis B.The overexpression (10–100,000 folds excess) of HBV genome-free subviral particles (SVPs) was found in acute and chronic hepatitis B patients [2], making HBsAg an detected biomarker. It is estimated that more than 257 million people have suffered from chronic hepatitis B infection worldwide, and almost 9 million people have died from liver failure, cirrhosis, hepatocellular carcinoma, and other related diseases caused by HBV [1]. Hepatitis B surface antigen (HBsAg) is the envelope protein of HBV, which is the most conventional serological marker for the screening, diagnosis, and antiviral therapy evaluation of hepatitis B. The overexpression (10–100,000 folds excess) of HBV genome-free subviral particles (SVPs) was found in acute and chronic hepatitis B patients [2], making HBsAg an detected biomarker. HBsAg serological tests for HBV include radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA), and electrochemiluminescence immunoassay. These methods are complex, time consuming, and require large equipment and professional technicians. Given the complex and dynamic variations of HBV, a single
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