論文

査読有り 国際誌
2021年

AuNP Coupled Rapid Flow-Through Dot-Blot Immuno-Assay for Enhanced Detection of SARS-CoV-2 Specific Nucleocapsid and Receptor Binding Domain IgG.

International journal of nanomedicine
  • Bijon Kumar Sil
  • Mohd Raeed Jamiruddin
  • Md Ahsanul Haq
  • Mohib Ullah Khondoker
  • Nowshin Jahan
  • Shahad Saif Khandker
  • Tamanna Ali
  • Mumtarin Jannat Oishee
  • Taku Kaitsuka
  • Masayasu Mie
  • Kazuhito Tomizawa
  • Eiry Kobatake
  • Mainul Haque
  • Nihad Adnan
  • 全て表示

16
開始ページ
4739
終了ページ
4753
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.2147/IJN.S313140

Background: Serological tests detecting severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) are widely used in seroprevalence studies and evaluating the efficacy of the vaccination program. Some of the widely used serological testing techniques are enzyme-linked immune-sorbent assay (ELISA), chemiluminescence immunoassay (CLIA), and lateral flow immunoassay (LFIA). However, these tests are plagued with low sensitivity or specificity, time-consuming, labor-intensive, and expensive. We developed a serological test implementing flow-through dot-blot assay (FT-DBA) for SARS-CoV-2 specific IgG detection, which provides enhanced sensitivity and specificity while being quick to perform and easy to use. Methods: SARS-CoV-2 antigens were immobilized on nitrocellulose membrane to capture human IgG, which was then detected with anti-human IgG conjugated gold nanoparticle (hIgG-AuNP). A total of 181 samples were analyzed in-house. Within which 35 were further evaluated in US FDA-approved CLIA Elecsys SARS-CoV-2 assay. The positive panel consisted of RT-qPCR positive samples from patients with both <14 days and >14 days from the onset of clinical symptoms. The negative panel contained samples collected from the pre-pandemic era dengue patients and healthy donors during the pandemic. Moreover, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of FT-DBA were evaluated against RT-qPCR positive sera. However, the overall efficacies were assessed with sera that seroconverted against either nucleocapsid (NCP) or receptor-binding domain (RBD). Results: In-house ELISA selected a total of 81 true seropositive and 100 seronegative samples. The sensitivity of samples with <14 days using FT-DBA was 94.7%, increasing to 100% for samples >14 days. The overall detection sensitivity and specificity were 98.8% and 98%, respectively, whereas the overall PPV and NPV were 99.6% and 99%. Moreover, comparative analysis between in-house ELISA assays and FT-DBA revealed clinical agreement of Cohen's Kappa value of 0.944. The FT-DBA showed sensitivity and specificity of 100% when compared with commercial CLIA kits. Conclusion: The assay can confirm past SARS-CoV-2 infection with high accuracy within 2 minutes compared to commercial CLIA or in-house ELISA. It can help track SARS-CoV-2 disease progression, population screening, and vaccination response. The ease of use of the assay without requiring any instruments while being semi-quantitative provides the avenue of its implementation in remote areas around the globe, where conventional serodiagnosis is not feasible.

リンク情報
DOI
https://doi.org/10.2147/IJN.S313140
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/34267520
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8277418
ID情報
  • DOI : 10.2147/IJN.S313140
  • PubMed ID : 34267520
  • PubMed Central 記事ID : PMC8277418

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