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An open source and convenient method for the wide-spread testing of COVID-19 using deep throat sputum samples.
Huang, Sunny C; Pak, Thomas K; Graber, Cameron P; Searby, Charles C; Liu, Guanghao; Marcy, Jennifer; Yaszemski, Alexandra K; Bedell, Kurt; Bui, Emily; Perlman, Stanley; Zhang, Qihong; Wang, Kai; Sheffield, Val C; Carter, Calvin S.
  • Huang SC; Medical Scientist Training Program / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Pak TK; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Graber CP; Medical Scientist Training Program / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Searby CC; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Liu G; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Marcy J; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Yaszemski AK; Department of Neurology, University of Iowa Hospitals and Clinics, Iowa City, IA, United States of America.
  • Bedell K; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Bui E; Department of Neurology / Mayo Clinic Graduate School of Biomedical Sciences, Mayo Clinic, Rochester, MN, United States of America.
  • Perlman S; Department of Microbiology and Immunology / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Zhang Q; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Wang K; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Sheffield VC; Department of Microbiology and Immunology / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
  • Carter CS; Stead Family Department of Pediatrics/ Division of Medical Genetics and Genomic / Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, United States of America.
PeerJ ; 10: e13277, 2022.
Article in English | MEDLINE | ID: covidwho-1934568
ABSTRACT
Importance The rise of novel, more infectious SARS-CoV-2 variants has made clear the need to rapidly deploy large-scale testing for COVID-19 to protect public health. However, testing remains limited due to shortages of personal protective equipment (PPE), naso- and oropharyngeal swabs, and healthcare workers. Simple test methods are needed to enhance COVID-19 screening. Here, we describe a simple, and inexpensive spit-test for COVID-19 screening called Patient Self-Collection of Sample-CoV2 (PSCS-CoV2).

Objective:

To evaluate an affordable and convenient test for COVID-19.

Methods:

The collection method relies on deep throat sputum (DTS) self-collected by the subject without the use of swabs, and was hence termed the Self-Collection of Sample for SARS-CoV-2 (abbreviated PSCS-CoV2). We used a phenol-chloroform extraction method for the viral RNA. We then tested for SARS-CoV-2 using real-time reverse transcription polymerase chain reaction with primers against at least two coding regions of the viral nucleocapsid protein (N1 and N2 or E) of SARS-CoV-2. We evaluted the sensitivity and specificity of our protocol. In addition we assess the limit of detection, and efficacy of our Viral Inactivating Solution. We also evaluated our protocol, and pooling strategy from volunteers on a local college campus.

Results:

We show that the PSCS-CoV2 method accurately identified 42 confirmed COVID-19 positives, which were confirmed through the nasopharyngeal swabbing method of an FDA approved testing facility. For samples negative for COVID-19, we show that the cycle threshold for N1, N2, and RP are similar between the PSCS-CoV2 and nasopharynx swab collection method (n = 30). We found a sensitivity of 100% (95% Confidence Interval [CI], 92-100) and specifity of 100% (95% CI, 89-100) for our PSCS-CoV2 method. We determined our protocol has a limit of detection of 1/10,000 for DTS from a COVID-19 patient. In addition, we show field data of the PSCS-CoV2 method on a college campus. Ten of the twelve volunteers (N1 < 30) that we tested as positive were subsequently tested positive by an independent laboratory. Finally, we show proof of concept of a pooling strategy to test for COVID-19, and recommend pool sizes of four if the positivity rate is less than 15%. Conclusion and Relevance We developed a DTS-based protocol for COVID-19 testing with high sensitivity and specificity. This protocol can be used by non-debilitated adults without the assistance of another adult, or by non-debilitated children with the assistance of a parent or guardian. We also discuss pooling strategies based on estimated positivity rates to help conserve resources, time, and increase throughput. The PSCS-CoV2 method can be a key component of community-wide efforts to slow the spread of COVID-19.
Keywords

Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies Topics: Variants Language: English Journal: PeerJ Year: 2022 Document Type: Article Affiliation country: Peerj.13277

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Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies Topics: Variants Language: English Journal: PeerJ Year: 2022 Document Type: Article Affiliation country: Peerj.13277