Development of a lateral flow immunoassay with HRP enhancement for spiked SARS-CoV-2 protein N detection in human saliva

Authors

  • Minh Hieu Vu
  • Doan Hong Ngoc Tran
  • Minh-Anh Dang-Trinh
  • Huynh Chan Khon*

Abstract

The Coronavirus (SARS-CoV-2) is an enveloped RNA virus that caused a dangerous COVID-19 pandemic following an outbreak in Wuhan, China in 2019. In response to the pandemic, the development of lateral flow assays (LFAs) has been crucial for the detection of viruses, commonly targeting the spike (S) or nucleocapsid (N) protein in nasopharyngeal swab (NS) specimens. COVID-19, caused by the SARS-CoV-2 virus, predominantly manifests as a respiratory tract infection-like illness, characterized by symptoms such as fever, dry cough, upper airway congestion, runny nose, sore throat, myalgia, headache, and exhaustion. This study presents the development of an LFA targeting the N protein to detect the coronavirus in saliva specimens, using a sandwich format. The use of 80 nm gold nanoparticles (AuNPs) has been investigated, and the application of horseradish peroxidase (HRP) and TMB substrate has been employed to enhance the limit of detection (LOD). The results demonstrate a significant 200-fold improvement in LOD, from 10 to 50 pg/ml, for N protein spiked in saliva samples after the application of HRP-TMB. This finding highlights an important advancement towards the utilization of saliva samples in diagnostic applications.

Keywords:

horseradish peroxidase, lateral flow assay, mixed-sized gold nanoparticles, SARS-CoV-2 nucleocapsid protein

DOI:

https://doi.org/10.31276/VJSTE.66(1).111-119

Classification number

3.5, 3.6

Author Biographies

Minh Hieu Vu

School of Biotechnology, International University, Vietnam National University - Ho Chi Minh City, Quarter 6, Linh Trung Ward, Thu Duc City, Ho Chi Minh City, Vietnam

Institute of Malariology, Parasitology and Entomology, Ho Chi Minh City, 699 Tran Hung Dao Street, Ward 1, District 5, Ho Chi Minh City, Vietnam

Doan Hong Ngoc Tran

Institute of Malariology, Parasitology and Entomology, Ho Chi Minh City, 699 Tran Hung Dao Street, Ward 1, District 5, Ho Chi Minh City, Vietnam

School of Biomedical Engineering, International University, Vietnam National University - Ho Chi Minh City,
Quarter 6, Linh Trung Ward, Thu Duc City, Ho Chi Minh City, Vietnam

Minh-Anh Dang-Trinh

Institute of Malariology, Parasitology and Entomology, Ho Chi Minh City, 699 Tran Hung Dao Street, Ward 1, District 5, Ho Chi Minh City, Vietnam

Huynh Chan Khon

School of Biomedical Engineering, International University, Vietnam National University - Ho Chi Minh City, Quarter 6, Linh Trung Ward, Thu Duc City, Ho Chi Minh City, Vietnam

Research Centre for Infectious Diseases, International University, Vietnam National University - Ho Chi Minh City, Quarter 6, Linh Trung Ward, Thu Duc City, Ho Chi Minh City, Vietnam

Vietnam National University - Ho Chi Minh City, Linh Trung Ward, Thu Duc City, Ho Chi Minh City, Vietnam

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Published

2024-03-15

Received 22 March 2023; revised 26 May 2023; accepted 21 June 2023

How to Cite

Minh Hieu Vu, Doan Hong Ngoc Tran, Minh-Anh Dang-Trinh, & Huynh Chan Khon. (2024). Development of a lateral flow immunoassay with HRP enhancement for spiked SARS-CoV-2 protein N detection in human saliva. Vietnam Journal of Science, Technology and Engineering, 66(1), 111-119. https://doi.org/10.31276/VJSTE.66(1).111-119

Issue

Section

Life Sciences