Ann Lab Med.  2018 Nov;38(6):578-584. 10.3343/alm.2018.38.6.578.

Development of a Rapid Automated Fluorescent Lateral Flow Immunoassay to Detect Hepatitis B Surface Antigen (HBsAg), Antibody to HBsAg, and Antibody to Hepatitis C

• Ryu JH
• Kwon M
• Moon JD
• Hwang MW
• Lee JM
• Park KH
• Yun SJ
• Bae HJ
• Choi A
• Lee H
• Jung B
• Jeong J
• Han K
• Kim Y
• Oh EJ

• KMID: 2429124
• DOI: http://doi.org/10.3343/alm.2018.38.6.578

Abstract

BACKGROUND
Accurate, rapid, and cost-effective screening tests for hepatitis B virus (HBV) and hepatitis C virus (HCV) infection may be useful in laboratories that cannot afford automated chemiluminescent immunoassays (CLIAs). We evaluated the diagnostic performance of a novel rapid automated fluorescent lateral flow immunoassay (LFIA).
METHODS
A fluorescent LFIA using a small bench-top fluorescence reader, Automated Fluorescent Immunoassay System (AFIAS; Boditech Med Inc., Chuncheon, Korea), was developed for qualitative detection of hepatitis B surface antigen (HBsAg), antibody to HBsAg (anti-HBs), and antibody to HCV (anti-HCV) within 20 minutes. We compared the diagnostic performance of AFIAS with that of automated CLIAs"”Elecsys (Roche Diagnostics GmbH, Penzberg, Germany) and ARCHITECT (Abbott Laboratories, Abbott Park, IL, USA)"”using 20 seroconversion panels and 3,500 clinical serum samples.
RESULTS
Evaluation with the seroconversion panels demonstrated that AFIAS had adequate sensitivity for HBsAg and anti-HCV detection. From the clinical samples, AFIAS sensitivity and specificity were 99.8% and 99.3% for the HBsAg test, 100.0% and 100.0% for the anti-HBs test, and 98.8% and 99.1% for the anti-HCV test, respectively. Its agreement rates with the Elecsys HBsAg, anti-HBs, and anti-HCV detection assays were 99.4%, 100.0%, and 99.0%, respectively. AFIAS detected all samples with HBsAg genotypes A-F and H and anti-HCV genotypes 1, 1a, 1b, 2a, 2b, 4, and 6. Cross-reactivity with other infections was not observed.
CONCLUSIONS
The AFIAS HBsAg, anti-HBs, and anti-HCV tests demonstrated diagnostic performance equivalent to current automated CLIAs. AFIAS could be used for a large-scale HBV or HCV screening in low-resource laboratories or low-to middle-income areas.

Keyword

HBsAg; Anti-HBs; Anti-HCV; Lateral flow immunoassay; Diagnostic performance

MeSH Terms

Fluorescence
Gangwon-do
Genotype
Hepacivirus
Hepatitis B Surface Antigens*
Hepatitis B virus
Hepatitis B*
Hepatitis C*
Hepatitis*
Immunoassay*
Mass Screening
Sensitivity and Specificity
Seroconversion
Hepatitis B Surface Antigens

Figure

• 

  Fig. 1 Schematic illustration of Automated Fluorescent Immunoassay System (AFIAS). (A) AFIAS-6 system and cartridge: AFIAS all-in-one cartridges are designed to optimize the structure and operating principle of the reader. The automated test process enables the performance of multiple simultaneous tests for six different samples. (B) Schematic representation of fluorescent signal detection by LFIA: AFIAS uses a sandwich immunodetection method. The cartridge contains a detection buffer including the dried reagent and monoclonal anti-chicken IgY as an internal control; both dried reagent and monoclonal anti-chicken IgY are labeled with europium chelate [Eu(III)]. When mixed with the sample, the detector binds to target molecules (viral antigens of antibodies) in the sample, forming antigen-antibody complexes. When the complexes migrate onto the nitrocellulose matrix, the other capture (antigen or antibodies) immobilized on the “Test line” forms a sandwich complex. The fluorescence-labeled anti-chicken IgY binds to the chicken IgY fixed to the “Control line.” The AFIAS scanner quantifies the analytes by measuring the fluorescence intensity on the test strip induced by a laser. The fluorescence detector has a fixed absorption wavelength of 333 nm and an emission wavelength of 613 nm, which are the standard wavelengths for the detection of europium [Eu(III)] chelate conjugates. Fluorescence intensity is proportional to the concentration of the target molecules in the samples. The result of the samples is given as “positive,” “negative,” or “indeterminate” in the form of a relative cut-off index (COI).

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