RT Journal Article
T1 A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection
A1 Reijns, Martin A. M.
A1 Thompson, Louise
A1 Acosta, Juan Carlos
A1 Black, Holly A.
A1 Sanchez-Luque, Francisco J.
A1 Diamond, Austin
A1 Parry, David A.
A1 Daniels, Alison
A1 O'Shea, Marie
A1 Uggenti, Carolina
A1 Sanchez, Maria C.
A1 O'Callaghan, Alan
A1 McNab, Michelle L. L.
A1 Adamowicz, Martyna
A1 Friman, Elias T.
A1 Hurd, Toby
A1 Jarman, Edward J.
A1 Chee, Frederic Li Mow
A1 Rainger, Jacqueline K.
A1 Walker, Marion
A1 Drake, Camilla
A1 Longman, Dasa
A1 Mordstein, Christine
A1 Warlow, Sophie J.
A1 McKay, Stewart
A1 Slater, Louise
A1 Ansari, Morad
A1 Tomlinson, Ian P. M.
A1 Moore, David
A1 Wilkinson, Nadine
A1 Shepherd, Jill
A1 Templeton, Kate
A1 Johannessen, Ingolfur
A1 Tait-Burkard, Christine
A1 Haas, Jürgen G.
A1 Gilbert, Nick
A1 Adams, Ian R.
A1 Jackson, Andrew P.
K1 COVID-19
K1 SARS-CoV-2
K1 Multiplex polymerase chain reaction
K1 Reacción en cadena de la polimerasa multiplex
K1 Reacción en cadena de la polimerasa de transcriptasa inversa
K1 Reverse transcriptase polymerase chain reaction
K1 ARN, viral
K1 RNA, viral
K1 Immunologic tests
K1 Pruebas inmunológicas
K1 Polymerase chain reaction
K1 Reacción en cadena de la polimerasa
K1 PCR
AB With the ongoing COVID-19 (Coronavirus Disease 2019) pandemic, caused by the novel coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), there is a need for sensitive, specific, and affordable diagnostic tests to identify infected individuals, not all of whom are symptomatic. The most sensitive test involves the detection of viral RNA using RT-qPCR (quantitative reverse transcription PCR), with many commercial kits now available for this purpose. However, these are expensive, and supply of such kits in sufficient numbers cannot always be guaranteed. We therefore developed a multiplex assay using well-established SARS-CoV-2 targets alongside a human cellular control (RPP30) and a viral spike-in control (Phocine Herpes Virus 1 [PhHV-1]), which monitor sample quality and nucleic acid extraction efficiency, respectively. Here, we establish that this test performs as well as widely used commercial assays, but at substantially reduced cost. Furthermore, we demonstrate >1,000-fold variability in material routinely collected by combined nose and throat swabbing and establish a statistically significant correlation between the detected level of human and SARS-CoV-2 nucleic acids. The inclusion of the human control probe in our assay therefore provides a quantitative measure of sample quality that could help reduce false-negative rates. We demonstrate the feasibility of establishing a robust RT-qPCR assay at approximately 10% of the cost of equivalent commercial assays, which could benefit low-resource environments and make high-volume testing affordable.
PB PLOS
YR 2020
FD 2020-12-15
LK http://hdl.handle.net/10668/3427
UL http://hdl.handle.net/10668/3427
LA en
NO Reijns MAM, Thompson L, Acosta JC, Black HA, Sanchez-Luque FJ, Diamond A, et al. A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection. PLoS Biol. 2020 Dec 15;18(12):e3001030.
DS RISalud
RD Apr 8, 2025