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DEVELOPMENT AND TESTING OF A LOW-COST INACTIVATION BUFFER THAT ALLOWS DIRECT SARS-COV-2 DETECTION IN SALIVA
Brandon Bustos-Garcia; Sylvia Garza-Manero; Nallely Cano-Dominguez; Dulce Maria Lopez Sanchez; Gonzalo Salgado-Montes de Oca; Alfonso Salgado-Aguayo; Felix Recillas-Targa; Santiago Avila-Rios; Julian Valdes.
  • Brandon Bustos-Garcia; Institute of Cellular Physiology (IFC), National Autonomous University of Mexico (UNAM)
  • Sylvia Garza-Manero; Institute of Cellular Physiology (IFC), National Autonomous University of Mexico (UNAM)
  • Nallely Cano-Dominguez; Institute of Cellular Physiology (IFC), National Autonomous University of Mexico
  • Dulce Maria Lopez Sanchez; Centre for Research in Infectious Diseases of the National Institute of Respiratory Diseases (CIENI/INER)
  • Gonzalo Salgado-Montes de Oca; Centre for Research in Infectious Diseases of the National Institute of Respiratory Diseases (CIENI/INER)
  • Alfonso Salgado-Aguayo; Centre for Research in Infectious Diseases of the National Institute of Respiratory Diseases (CIENI/INER)
  • Felix Recillas-Targa; Institute of Cellular Physiology (IFC), National Autonomous University of Mexico
  • Santiago Avila-Rios; National Institute of Respiratory Diseases
  • Julian Valdes; Institute of Cellular Physiology (IFC), National Autonomous University of Mexico
Preprint in English | medRxiv | ID: ppmedrxiv-21266918
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ABSTRACT
Massive testing is a cornerstone in efforts to effectively track infections and stop COVID-19 transmission, including places where good vaccination coverage has been achieved. However, SARS-CoV-2 testing by RT-qPCR requires specialized personnel, protection equipment, commercial kits, and dedicated facilities, which represent significant challenges for massive testing implementation in resource-limited settings. It is therefore important to develop testing protocols that facilitate implementation and are inexpensive, fast, and sufficiently sensitive. In this work, we optimized the composition of a buffer (PKTP) containing a protease, a detergent, and an RNase inhibitor, that is compatible with the RT-qPCR chemistry, allowing for direct testing of SARS-CoV-2 from saliva in an RNA extraction-independent manner. This buffer is compatible with heat-inactivation reducing the biohazard risk of handling the samples. We assessed the PKTP buffer performance in comparison to the RNA-extraction-based protocol of the US Centers for Disease Control and Prevention in saliva samples from 70 COVID-19 patients finding a good sensitivity (82.2% for the N1 and 84.4% for the N2 target, respectively) and correlations (R=0.77, p<0.001 for N1, and R=0.78, p<0.001 for N2). We also propose an auto-collection protocol for saliva samples and a multiplex reaction to reduce the number of PCR reactions per patient and further reduce overall costs while maintaining diagnostic standards in favor of massive testing.
Full text: Available Collection: Preprints Database: medRxiv Document Type: Preprint Language: English Year: 2021

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Full text: Available Collection: Preprints Database: medRxiv Document Type: Preprint Language: English Year: 2021
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