TY - JOUR
T1 - Validation of a molecular assay to detect SARS-CoV-2 in saliva
AU - Pitman, Janet L.
AU - Morris, Arthur J.
AU - Grice, Stephen
AU - Walsh, Joseph T.
AU - Wang, Leyi
AU - Burke, Martin D.
AU - Dixon-McIver, Amanda
N1 - Funding Information:
Dr Walsh reports grants from National Institutes for Health (NIBIB) and from the Rockefeller Foundation during the conduct of the study. He also reports that he is on the Board of Managers for SHIELD T3, an LLC whose mission is to provide SARS-CoV-2 tests based upon the technology described in this manuscript, and that he oversees SHIELD Illinois, a group within the University of Illinois that provides SARS-CoV-2 testing across the state of Illinois based upon the technology described in this manuscript. His remuneration is not supplemented by either the SHIELD T3 or SHIELD Illinois activities. Dr Pitman reports other from Rako Science during the conduct of the study. Dr Dixon-McIver reports other from Rako Science outside the submitted work. Dr Morris reports other from IGENZ outside the submitted work. Dr Grice reports personal fees from Rako Science outside the submitted work, and that Rako Science has licensed trade secrets related to the covidSHIELD protocol. Dr Wang reports they have a patent saliva-based molecular testing for SARS-CoV-2 pending to Diana Rose E Ranoa, Robin L Holland, Fadi G Alnaji, Kelsie J Green, Leyi Wang, Christopher B Brooke, Martin D Burke, Timothy M Fan, Paul J Hergenrother. Acknowledgements: The authors acknowledge Leticia Castro (School of Biological Sciences, Victoria University of Wellington, New Zealand) and Rebecca Perwick, Bronwyn Neumann, Akshay Nandan Kumar and Lili Jiang (IGENZ, Auckland, New Zealand) for their technical expertise in performing the validation testing. We also acknowledge Dr Diana Ranoa for sample matching between the laboratories. The authors thank Dr Gary McAuliffe for reviewing the validation protocol. Author information: Janet L Pitman: Associate Professor, School of Biological Sciences, Victoria University of Wellington, Kelburn Parade, Wellington. Arthur J Morris: Clinical Microbiologist, Auckland. Stephen Grice: Director, Rako Science Ltd, Level 7, 76 Manners Street, Te Aro, Wellington. Joseph T Walsh: Office of the Vice President for Economic Development and Innovation, University of Illinois System, Urbana, IL, USA. Leyi Wang: Clinical Assistant Professor, Veterinary Diagnostic Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL, USA. Martin D Burke: Professor, Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA. Amanda Dixon-McIver: Laboratory Director, IGENZ Limited, Auckland. Corresponding author: Janet Pitman, School of Biological Sciences, Victoria University of Wellington, PO Box 600, Kelburn Parade, Wellington 6140 janet.pitman@vuw.ac.nz URL: x
Funding Information:
The authors acknowledge Leticia Castro (School of Biological Sciences, Victoria University of Wellington, New Zealand) and Rebecca Perwick, Bronwyn Neumann, Akshay Nandan Kumar and Lili Jiang (IGENZ, Auckland, New Zealand) for their technical expertise in performing the validation testing. We also acknowledge Dr Diana Ranoa for sample matching between the laboratories. The authors thank Dr Gary McAuliffe for reviewing the validation protocol.
Publisher Copyright:
© NZMA
PY - 2021/12/17
Y1 - 2021/12/17
N2 - AIM: To validate a reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) assay to detect SARS-CoV-2 in saliva in two independent Aotearoa New Zealand laboratories. METHODS: An RT-qPCR assay developed at University of Illinois Urbana-Champaign, USA, was validated in two New Zealand laboratories. Analytical measures, such as limit of detection (LOD) and cross-reactivity, were performed. One hundred and forty-seven saliva samples, each paired with a contemporaneously collected nasal swab, mainly of nasopharyngeal origin, were received. Positive (N=33) and negative (N=114) samples were tested blindly in each laboratory. Diagnostic sensitivity and specificity were then calculated. RESULTS: The LOD was <0.75 copy per µL and no cross-reactivity with MERS-CoV was detected. There was complete concordance between laboratories for all saliva samples with the quantification cycle values for all three genes in close agreement. Saliva had 98.7% concordance with paired nasal samples: and a sensitivity, specificity and accuracy of 97.0%, 99.1% and 99.1%, respectively. CONCLUSION: This saliva RT-qPCR assay produces reproducible results with a low LOD. High sensitivity and specificity make it a reliable option for SARS-CoV-2 testing, including for asymptomatic people requiring regular screening.
AB - AIM: To validate a reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) assay to detect SARS-CoV-2 in saliva in two independent Aotearoa New Zealand laboratories. METHODS: An RT-qPCR assay developed at University of Illinois Urbana-Champaign, USA, was validated in two New Zealand laboratories. Analytical measures, such as limit of detection (LOD) and cross-reactivity, were performed. One hundred and forty-seven saliva samples, each paired with a contemporaneously collected nasal swab, mainly of nasopharyngeal origin, were received. Positive (N=33) and negative (N=114) samples were tested blindly in each laboratory. Diagnostic sensitivity and specificity were then calculated. RESULTS: The LOD was <0.75 copy per µL and no cross-reactivity with MERS-CoV was detected. There was complete concordance between laboratories for all saliva samples with the quantification cycle values for all three genes in close agreement. Saliva had 98.7% concordance with paired nasal samples: and a sensitivity, specificity and accuracy of 97.0%, 99.1% and 99.1%, respectively. CONCLUSION: This saliva RT-qPCR assay produces reproducible results with a low LOD. High sensitivity and specificity make it a reliable option for SARS-CoV-2 testing, including for asymptomatic people requiring regular screening.
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M3 - Article
C2 - 35728108
AN - SCOPUS:85126646670
VL - 134
SP - 14
EP - 27
JO - New Zealand Medical Journal
JF - New Zealand Medical Journal
SN - 0028-8446
IS - 1547
ER -