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Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source Pipetting Robots
JoVE Journal
Imunologia e Infecção
This content is Free Access.
JoVE Journal Imunologia e Infecção
Efficient SARS-CoV-2 Quantitative Reverse Transcriptase PCR Saliva Diagnostic Strategy utilizing Open-Source Pipetting Robots
DOI:

11:11 min

February 11, 2022

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Capítulos

  • 00:04Introduction
  • 00:56Sample Intake, Heat Treatment, and Sample Assignment
  • 01:52Operating Sample Loading Robots
  • 04:09Manual Sample Loading
  • 04:52Addition of Controls to Testing Plates
  • 05:54Performing RT-qPCR and Determining Plate Validity
  • 07:00Interpreting Sample Results
  • 08:08Results: RT-qPCR Detection of N1 (SARS-CoV-2) Synthetic RNA and P1 (Hs_RPP30) Synthetic DNA, Comparison Between Manual and Automated Saliva Transfer N1 Ct Values, and Evaluation of Heat Treatment Methods for Viscosity Reduction in Saliva
  • 10:26Conclusion

Summary

Tadução automática

The protocol describes a SARS-CoV-2 diagnostic method that utilizes open-source automation to perform RT-qPCR molecular testing of saliva samples. This scalable approach can be applied to clinical public health surveillance as well as to increase the capacity of smaller university laboratories.

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