Imperial College London
Dr Philippa C. May

Dr Philippa C. May

Faculty of Medicine, Department of Immunology and Inflammation

Visiting Researcher
 
 
 
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Contact

 

philippa.may

 
 
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Location

 

4N5Commonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Rowan:2021:10.1016/j.jviromet.2021.114174,
author = {Rowan, AG and May, P and Badhan, A and Herrera, C and Watber, P and Penn, R and Crone, MA and Storch, M and Garson, JA and McClure, M and Freemont, PS and Madona, P and Randell, P and Taylor, GP},
doi = {10.1016/j.jviromet.2021.114174},
journal = {Journal of Virological Methods},
pages = {1--7},
title = {Optimized protocol for a quantitative SARS-CoV-2 duplex RT-qPCR assay with internal human sample sufficiency control.},
url = {http://dx.doi.org/10.1016/j.jviromet.2021.114174},
volume = {294},
year = {2021}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - There is growing evidence that measurement of SARS-CoV-2 viral copy number can inform clinical and public health management of SARS-CoV-2 carriers and COVID-19 patients. Here we show that quantification of SARS-CoV-2 is feasible in a clinical setting, using a duplex RT-qPCR assay which targets both the E gene (Charité assay) and a human RNA transcript, RNase P (CDC assay) as an internal sample sufficiency control. Samples in which RNase P is not amplified indicate that sample degradation has occurred, PCR inhibitors are present, RNA extraction has failed or swabbing technique was insufficient. This important internal control reveals that 2.4% of nasopharyngeal swabs (15/618 samples) are inadequate for SARS-CoV-2 testing which, if not identified, could result in false negative results. We show that our assay is linear across at least 7 logs and is highly reproducible, enabling the conversion of Cq values to viral copy numbers using a standard curve. Furthermore, the SARS-CoV-2 copy number was independent of the RNase P copy number indicating that the per-swab viral copy number is not dependent on sampling- further allowing comparisons between samples. The ability to quantify SARS-CoV-2 viral copy number will provide an important opportunity for viral burden-guided public health and clinical decision making.
AU  - Rowan,AG
AU  - May,P
AU  - Badhan,A
AU  - Herrera,C
AU  - Watber,P
AU  - Penn,R
AU  - Crone,MA
AU  - Storch,M
AU  - Garson,JA
AU  - McClure,M
AU  - Freemont,PS
AU  - Madona,P
AU  - Randell,P
AU  - Taylor,GP
DO  - 10.1016/j.jviromet.2021.114174
EP  - 7
PY  - 2021///
SN  - 0166-0934
SP  - 1
TI  - Optimized protocol for a quantitative SARS-CoV-2 duplex RT-qPCR assay with internal human sample sufficiency control.
T2  - Journal of Virological Methods
UR  - http://dx.doi.org/10.1016/j.jviromet.2021.114174
UR  - https://www.ncbi.nlm.nih.gov/pubmed/33984396
UR  - https://www.sciencedirect.com/science/article/pii/S0166093421001130?via%3Dihub
UR  - http://hdl.handle.net/10044/1/88654
VL  - 294
ER  -
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