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Robotic RNA extraction for SARS-CoV-2 surveillance using saliva samples.
Hamilton, Jennifer R; Stahl, Elizabeth C; Tsuchida, Connor A; Lin-Shiao, Enrique; Tsui, C Kimberly; Pestal, Kathleen; Gildea, Holly K; Witkowsky, Lea B; Moehle, Erica A; McDevitt, Shana L; McElroy, Matthew; Keller, Amanda; Sylvain, Iman; Hirsh, Ariana; Ciling, Alison; Ehrenberg, Alexander J; Ringeisen, Bradley R; Huberty, Garth; Urnov, Fyodor D; Giannikopoulos, Petros; Doudna, Jennifer A.
Afiliação
  • Hamilton JR; University of California, Berkeley, Berkeley, CA, USA.
  • Stahl EC; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Tsuchida CA; University of California, Berkeley, Berkeley, CA, USA.
  • Lin-Shiao E; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Tsui CK; University of California, Berkeley, Berkeley, CA, USA.
  • Pestal K; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Gildea HK; University of California, Berkeley, Berkeley, CA, USA.
  • Witkowsky LB; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Moehle EA; University of California, Berkeley, Berkeley, CA, USA.
  • McDevitt SL; University of California, Berkeley, Berkeley, CA, USA.
  • McElroy M; University of California, Berkeley, Berkeley, CA, USA.
  • Keller A; University of California, Berkeley, Berkeley, CA, USA.
  • Sylvain I; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Hirsh A; University of California, Berkeley, Berkeley, CA, USA.
  • Ciling A; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Ehrenberg AJ; University of California, Berkeley, Berkeley, CA, USA.
  • Ringeisen BR; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Huberty G; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Urnov FD; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • Giannikopoulos P; University of California, Berkeley, Berkeley, CA, USA.
  • Doudna JA; Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
medRxiv ; 2021 Jan 29.
Article em En | MEDLINE | ID: mdl-33532798
ABSTRACT
Saliva is an attractive specimen type for asymptomatic surveillance of COVID-19 in large populations due to its ease of collection and its demonstrated utility for detecting RNA from SARS-CoV-2. Multiple saliva-based viral detection protocols use a direct-to-RT-qPCR approach that eliminates nucleic acid extraction but can reduce viral RNA detection sensitivity. To improve test sensitivity while maintaining speed, we developed a robotic nucleic acid extraction method for detecting SARS-CoV-2 RNA in saliva samples with high throughput. Using this assay, the Free Asymptomatic Saliva Testing (IGI-FAST) research study on the UC Berkeley campus conducted 11,971 tests on supervised self-collected saliva samples and identified rare positive specimens containing SARS-CoV-2 RNA during a time of low infection prevalence. In an attempt to increase testing capacity, we further adapted our robotic extraction assay to process pooled saliva samples. We also benchmarked our assay against the gold standard, nasopharyngeal swab specimens. Finally, we designed and validated a RT-qPCR test suitable for saliva self-collection. These results establish a robotic extraction-based procedure for rapid PCR-based saliva testing that is suitable for samples from both symptomatic and asymptomatic individuals.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Risk_factors_studies / Screening_studies Idioma: En Ano de publicação: 2021 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Tipo de estudo: Risk_factors_studies / Screening_studies Idioma: En Ano de publicação: 2021 Tipo de documento: Article