tetano
Editor, Senior Moderator
J Biomol Tech
. 2021 Sep;32(3):148-157.
doi: 10.7171/jbt.21-3203-010.
Rapid, Affordable, and Scalable SARS-CoV-2 Detection From Saliva
Andrew Hayden[SUP] 1 [/SUP], Marcy Kuentzel[SUP] 1 [/SUP], Sridar V Chittur[SUP] 1 2 [/SUP]
Affiliations
Abstract
Here we present an inexpensive, rapid, and robust reverse-transcription loop-mediated isothermal amplification (RT-LAMP)-based severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection method that is easily scalable, enabling point-of-care facilities and clinical labs to determine results from patients' saliva directly in 30 minutes for less than $2 per reaction. The method uses a novel combination of widely available reagents that can be prepared in bulk, plated, and frozen and remain stable until samples are received. This innovation dramatically reduces preparation time, enabling high-throughput automation and testing with time to results (including setup) in less than 1 hour for 96 patient samples simultaneously when using a 384-well format. By using a dual reporter (phenol red pH indicator for end-point detection and SYTO-9 fluorescent dye for real time), the assay also provides internal validation of results and redundancy in the event of an instrument malfunction.
Keywords: Fluorescent LAMP; RT-LAMP; extraction-free detection; saliva detection.
. 2021 Sep;32(3):148-157.
doi: 10.7171/jbt.21-3203-010.
Rapid, Affordable, and Scalable SARS-CoV-2 Detection From Saliva
Andrew Hayden[SUP] 1 [/SUP], Marcy Kuentzel[SUP] 1 [/SUP], Sridar V Chittur[SUP] 1 2 [/SUP]
Affiliations
- PMID: 35027872
- PMCID: PMC8730515
- DOI: 10.7171/jbt.21-3203-010
Abstract
Here we present an inexpensive, rapid, and robust reverse-transcription loop-mediated isothermal amplification (RT-LAMP)-based severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection method that is easily scalable, enabling point-of-care facilities and clinical labs to determine results from patients' saliva directly in 30 minutes for less than $2 per reaction. The method uses a novel combination of widely available reagents that can be prepared in bulk, plated, and frozen and remain stable until samples are received. This innovation dramatically reduces preparation time, enabling high-throughput automation and testing with time to results (including setup) in less than 1 hour for 96 patient samples simultaneously when using a 384-well format. By using a dual reporter (phenol red pH indicator for end-point detection and SYTO-9 fluorescent dye for real time), the assay also provides internal validation of results and redundancy in the event of an instrument malfunction.
Keywords: Fluorescent LAMP; RT-LAMP; extraction-free detection; saliva detection.