Latticed Gold Nanoparticle Conjugation via Monomeric Streptavidin in Lateral Flow Assay for Detection of Autoantibody to Interferon-Gamma

Weeraya Thongkum; Umpa Yasamut; Koollawat Chupradit; Supachai Sakkhachornphop; Jiraprapa Wipasa; Kanokporn Sornsuwan; On-Anong Juntit; Rawiwan Pornprasit; Wanwisa Thongkamwitoon; Jirapan Chaichanan; Jaruwan Khaoplab; Chonnikarn Chanpradab; Watchara Kasinrerk; Chatchai Tayapiwatana

doi:10.3390/diagnostics11060987

Latticed Gold Nanoparticle Conjugation via Monomeric Streptavidin in Lateral Flow Assay for Detection of Autoantibody to Interferon-Gamma

Diagnostics (Basel). 2021 May 29;11(6):987. doi: 10.3390/diagnostics11060987.

Authors

Weeraya Thongkum^{1

2

3}, Umpa Yasamut^{1

2

3}, Koollawat Chupradit^{1

2}, Supachai Sakkhachornphop^{2

4}, Jiraprapa Wipasa⁴, Kanokporn Sornsuwan^{1

2}, On-Anong Juntit^{1

2}, Rawiwan Pornprasit⁵, Wanwisa Thongkamwitoon⁵, Jirapan Chaichanan⁵, Jaruwan Khaoplab⁵, Chonnikarn Chanpradab⁵, Watchara Kasinrerk^{1

3}, Chatchai Tayapiwatana^{1

2

3}

Affiliations

¹ Division of Clinical Immunology, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
² Center of Biomolecular Therapy and Diagnostic, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
³ Center of Innovative Immunodiagnostic Development, Department of Medical Technology, Faculty of Associated Medical Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
⁴ Research Institute for Health Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
⁵ Bio Innovation Building, Mahidol University, Nakhon Pathom 73170, Thailand.

Abstract

Adult-onset immunodeficiency syndrome (AOID) patients with autoantibodies (autoAbs) against interferon-gamma (IFN-γ) generally suffer from recurrent and recalcitrant disseminated non-tuberculous mycobacterial diseases. Since the early stages of AOID do not present specific symptoms, diagnosis and treatment of the condition are not practical. A simplified diagnostic method for differentiating AOID from other immunodeficiencies, such as HIV infection, was created. Anti-IFN-γ is generally identified using enzyme-linked immunosorbent assay (ELISA), which involves an instrument and a cumbersome process. Recombinant IFN-γ indirectly conjugated to colloidal gold was used in the modified immunochromatographic (IC) strips. The biotinylated-IFN-γ was incorporated with colloidal-gold-labeled 6HIS-maltose binding protein-monomeric streptavidin (^6HISMBP-mSA) and absorbed at the conjugate pad. The efficacy of the IC strip upon applying an anti-IFN-γ autoAb cut-off ELISA titer of 2500, the sensitivity and specificity were 84% and 90.24%, respectively. When a cut-off ELISA titer of 500 was applied, the sensitivity and specificity were 73.52% and 100%, respectively.

Keywords: anti-interferon-γ autoantibody; conventional indirect ELISA; immunochromatographic strip test; interferon-γ.

Abstract

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