TY - JOUR
T1 - Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer
AU - Erster, Oran
AU - Shkedi, Omer
AU - Benedek, Gil
AU - Zilber, Eyal
AU - Varkovitzky, Itay
AU - Shirazi, Rachel
AU - Shorka, Dorit Oriya
AU - Cohen, Yuval
AU - Bar, Tzahi
AU - Yechieli, Rafi
AU - Oikawa, Michal Tepperberg
AU - Venkert, Dana
AU - Linial, Michal
AU - Oiknine-Djian, Esther
AU - Mandelboim, Michal
AU - Livneh, Zvi
AU - Shenhav-Saltzman, Gilat
AU - Mendelson, Ella
AU - Wolf, Dana
AU - Szwarcwort-Cohen, Moran
AU - Mor, Orna
AU - Lewis, Yair
AU - Zeevi, Danny
N1 - Publisher Copyright:
© 2021 Erster et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2021/3
Y1 - 2021/3
N2 - Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preservative, thus inactivating the virus immediately after sampling. We tested 79 COVID-19 patients and 20 healthy controls. We collected two samples (nasopharyngeal swabs) from each participant: One swab was inserted into a test tube with Viral Transport Medium (VTM), following the standard guideline used as the recommended method for sample collection; the other swab was inserted into a lysis buffer supplemented with nucleic acid stabilization mix (coined NSLB). We found that RT-qPCR tests of patients were significantly more sensitive with NSLB sampling, reaching detection threshold 2.1±0.6 (Mean±SE) PCR cycles earlier then VTM samples from the same patient. We show that this improvement is most likely since NSLB samples are not diluted in lysis buffer before RNA extraction. Re-extracting RNA from NSLB samples after 72 hours at room temperature did not affect the sensitivity of detection, demonstrating that NSLB allows for long periods of sample preservation without special cooling equipment. We also show that swirling the swab in NSLB and discarding it did not reduce sensitivity compared to retaining the swab in the tube, thus allowing improved automation of COVID-19 tests. Overall, we show that using NSLB instead of VTM can improve the sensitivity, safety, and rapidity of COVID-19 tests at a time most needed.
AB - Conducting numerous, rapid, and reliable PCR tests for SARS-CoV-2 is essential for our ability to monitor and control the current COVID-19 pandemic. Here, we tested the sensitivity and efficiency of SARS-CoV-2 detection in clinical samples collected directly into a mix of lysis buffer and RNA preservative, thus inactivating the virus immediately after sampling. We tested 79 COVID-19 patients and 20 healthy controls. We collected two samples (nasopharyngeal swabs) from each participant: One swab was inserted into a test tube with Viral Transport Medium (VTM), following the standard guideline used as the recommended method for sample collection; the other swab was inserted into a lysis buffer supplemented with nucleic acid stabilization mix (coined NSLB). We found that RT-qPCR tests of patients were significantly more sensitive with NSLB sampling, reaching detection threshold 2.1±0.6 (Mean±SE) PCR cycles earlier then VTM samples from the same patient. We show that this improvement is most likely since NSLB samples are not diluted in lysis buffer before RNA extraction. Re-extracting RNA from NSLB samples after 72 hours at room temperature did not affect the sensitivity of detection, demonstrating that NSLB allows for long periods of sample preservation without special cooling equipment. We also show that swirling the swab in NSLB and discarding it did not reduce sensitivity compared to retaining the swab in the tube, thus allowing improved automation of COVID-19 tests. Overall, we show that using NSLB instead of VTM can improve the sensitivity, safety, and rapidity of COVID-19 tests at a time most needed.
UR - http://www.scopus.com/inward/record.url?scp=85103533668&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0249149
DO - 10.1371/journal.pone.0249149
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C2 - 33784369
AN - SCOPUS:85103533668
SN - 1932-6203
VL - 16
JO - PLoS ONE
JF - PLoS ONE
IS - 3 March
M1 - e0249149
ER -
