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Pretreatment Methods for Human Nasopharyngeal Swabs to Increase the Signal to Noise Ratio of High Sensitivity Immunoassays.
Chen, Hui; Feng, Sheng; Zhou, Wan; Li, Zhao; Richard-Greenblatt, Melissa; Wang, Ping.
  • Chen H; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • Feng S; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • Zhou W; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • Li Z; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • Richard-Greenblatt M; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
  • Wang P; Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, United States.
ACS Meas Sci Au ; 2(5): 414-421, 2022 Oct 19.
Article in English | MEDLINE | ID: covidwho-2185512
ABSTRACT
Mucous samples collected through nasopharyngeal (NP) swabs are considered gold standard specimens for the detection of respiratory pathogens. Matrices of these highly viscous samples often cause significant background noises in immunoassays, especially immunoassays with high sensitivity. We demonstrated such nonspecific background signals in both a chemiluminescence enzyme-linked immunosorbent assay (ELISA) and a novel highly sensitive immunoassay called Microbubbling SARS-CoV-2 Antigen Assay (MSAA). We developed and demonstrated the effectiveness of two quick sample pretreatment methods, filtration and preadsorption, to decrease nonspecific signals and increase the signal-to-noise ratio (SNR). Using these pretreatment methods, the SNR (at 3.6 × 104 copies/mL of inactivated SARS-CoV-2) was increased by 42.4-fold (95% CI 41.0-43.8) and 67.1-fold (95% CI 57.9-76.3) in the MSAA, and 1.3-fold (95% CI 0.9-1.7) and 1.8-fold (95% CI 1.6-2.0) in the chemiluminescence ELISA assay. Sample pretreatment methods developed in this study are broadly adaptable for the development of immunoassays for highly viscous samples.

Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study Language: English Journal: ACS Meas Sci Au Year: 2022 Document Type: Article Affiliation country: Acsmeasuresciau.2c00024

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study Language: English Journal: ACS Meas Sci Au Year: 2022 Document Type: Article Affiliation country: Acsmeasuresciau.2c00024