Amplified parallel antigen rapid test for point-of-care salivary detection of SARS-CoV-2 with improved sensitivity

Mikrochim Acta. 2021 Dec 6;189(1):14. doi: 10.1007/s00604-021-05113-4.


In the ongoing COVID-19 pandemic, simple, rapid, point-of-care tests not requiring trained personnel for primary care testing are essential. Saliva-based antigen rapid tests (ARTs) can fulfil this need, but these tests require overnight-fasted samples; without which independent studies have demonstrated sensitivities of only 11.7 to 23.1%. Herein, we report an Amplified Parallel ART (AP-ART) with sensitivity above 90%, even with non-fasted samples. The virus was captured multimodally, using both anti-spike protein antibodies and Angiotensin Converting Enzyme 2 (ACE2) protein. It also featured two parallel flow channels. The first contained spike protein binding gold nanoparticles which produced a visible red line upon encountering the virus. The second contained signal amplifying nanoparticles that complex with the former and amplify the signal without any linker. Compared to existing dual gold amplification techniques, a limit of detection of one order of magnitude lower was achieved (0.0064 ng·mL-1). AP-ART performance in detecting SARS-CoV-2 in saliva of COVID-19 patients was investigated using a case-control study (139 participants enrolled and 162 saliva samples tested). Unlike commercially available ARTs, the sensitivity of AP-ART was maintained even when non-fasting saliva was used. Compared to the gold standard reverse transcription-polymerase chain reaction testing on nasopharyngeal samples, non-fasting saliva tested on AP-ART showed a sensitivity of 97.0% (95% CI: 84.7-99.8); without amplification, the sensitivity was 72.7% (95% CI: 83.7-94.8). Thus, AP-ART has the potential to be developed for point-of-care testing, which may be particularly important in resource-limited settings, and for early diagnosis to initiate newly approved therapies to reduce COVID-19 severity.

Keywords: Antigen rapid test; COVID-19; Gold nanoparticles; Parallel flow amplification; Point-of-care detection; SARS-CoV-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / analysis*
  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • Case-Control Studies
  • Gold / chemistry
  • Immunoassay / instrumentation
  • Immunoassay / methods
  • Metal Nanoparticles / chemistry
  • Point-of-Care Testing*
  • SARS-CoV-2 / isolation & purification
  • SARS-CoV-2 / metabolism
  • Saliva / virology*
  • Sensitivity and Specificity


  • Antigens
  • Gold