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Detection of influenza virus by a biosensor based on the method combining electrochemiluminescence on binary SAMs
Yumi Katayama1, Takayuki Ohgi1, Yoshiharu Mitoma1
1Department of Environmental Sciences, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, 562 Nanatsuka, Shobara, Hiroshima, 727-0023, Japan.
Insights
This study presents a novel electrochemiluminescence immunosensor for detecting influenza A (H1N1). The developed biosensor offers significantly improved sensitivity and accuracy compared to existing methods for influenza virus detection.
Area of Science:
- Biosensors
- Immunosensors
- Electrochemiluminescence (ECL)
- Influenza Virus Detection
Background:
- Current point-of-care testing (POCT) for influenza diagnosis suffers from low sensitivity, leading to false negatives.
- There is a critical need for highly sensitive and accurate diagnostic methods for influenza virus A (H1N1).
Purpose of the Study:
- To develop and validate an electrochemiluminescence (ECL) immunosensor for sensitive and accurate detection of influenza virus A (H1N1).
- To improve the sensitivity and accuracy of influenza virus detection by optimizing sensor surface chemistry.
Main Methods:
- An immunosensor was designed using immunoliposomes encapsulating a tris(2,2'-bipyridyl)ruthenium(II) complex.
- Detection involved a competitive immunoassay utilizing a hemagglutinin (HA) peptide immobilized on self-assembled monolayers (SAMs).
- Binary SAMs, composed of 3,3'-dithiodipropionic acid and 1-hexanethiol, were employed to enhance SAMs coverage on a gold electrode.
Main Results:
- The use of binary SAMs significantly improved electrode surface coverage and reduced background ECL signals.
- The optimized method successfully detected influenza virus A (H1N1) with enhanced sensitivity and accuracy.
- Detection range achieved was from 2.7 × 10^2 to 2.7 × 10^3 PFU/mL, outperforming POCT and ELISA.
Conclusions:
- The developed ECL immunosensor demonstrates superior performance for influenza virus A (H1N1) detection.
- This advanced biosensor offers a promising alternative for rapid, sensitive, and accurate influenza diagnostics.
Abstract:
Recently, point of care testing (POCT) used for diagnosis of influenza infection has a problem showing false negative diagnosis because of the low sensitivity. We would like to report detection of influenza virus A (H1N1) by an immunosensor based on electrochemiluminescence (ECL) that uses an immunoliposome encapsulating tris(2,2'-bipyridyl)ruthenium(II) complex. By using the sensor, we could detect the virus that competed with hemagglutinin (HA) peptide immobilized on self-assembled monolayers (SAMs) in immunoreaction of the antibody bound on the surface of liposome. The HA peptide was 19 mer (TGLRNGITNKVNSVIEKAA). We demonstrated great improvement of sensitivity and accuracy by introducing binary SAMs instead of mono SAMs. The binary SAMs was prepared from 3,3'-dithiodipropionic acid and 1-hexanethiol. Use of the binary SAMs enabled to increase the SAMs coverage on Au electrode; the fact was confirmed by observation of the cathodic desorption currents. By using such an electrode, first the detection method of BSA was optimized to lower ECL background signal. Then we applied the method to the detection of influenza virus. We could successfully detect the virus with higher sensitivity compared with that by POCT and ELISA. The detection range was from a concentration of 2.7 × 10(2) to 2.7 × 10(3) PFU/mL.
