Improved sensitivity, safety, and rapidity of COVID-19 tests by replacing viral storage solution with lysis buffer.


Journal

PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081

Informations de publication

Date de publication:
2021
Historique:
received: 08 12 2020
accepted: 12 03 2021
entrez: 30 3 2021
pubmed: 31 3 2021
medline: 10 4 2021
Statut: epublish

Résumé

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.

Identifiants

pubmed: 33784369
doi: 10.1371/journal.pone.0249149
pii: PONE-D-20-38567
pmc: PMC8009371
doi:

Substances chimiques

Buffers 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

e0249149

Déclaration de conflit d'intérêts

The authors have declared that no competing interests exist.

Références

Behav Res Methods. 2009 Nov;41(4):1149-60
pubmed: 19897823
Euro Surveill. 2020 Apr;25(14):
pubmed: 32290902
J Med Virol. 2020 Oct;92(10):1755-1756
pubmed: 32270882
Lancet Infect Dis. 2020 Nov;20(11):1231-1232
pubmed: 32530425
PLoS One. 2014 Sep 02;9(9):e105786
pubmed: 25181250
Euro Surveill. 2020 Aug;25(32):
pubmed: 32794447
Euro Surveill. 2020 Dec;25(50):
pubmed: 33334398
Nat Commun. 2020 Sep 23;11(1):4812
pubmed: 32968075

Auteurs

Oran Erster (O)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.

Omer Shkedi (O)

Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.

Gil Benedek (G)

Hadassah Medical Center, Jerusalem, Israel.

Eyal Zilber (E)

Department of Internal Medicine, Sheba Medical Center, Tel Hashomer, Ramat-Gan, Israel.

Itay Varkovitzky (I)

Directorate of Defense Research & Development, Israeli Ministry of Defense, Tel Aviv, Israel.

Rachel Shirazi (R)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.

Dorit Oriya Shorka (D)

Hadassah Medical Center, Jerusalem, Israel.

Yuval Cohen (Y)

Directorate of Defense Research & Development, Israeli Ministry of Defense, Tel Aviv, Israel.

Tzahi Bar (T)

Independent Researcher, Israel.

Rafi Yechieli (R)

Independent Researcher, Israel.

Michal Tepperberg Oikawa (M)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.

Dana Venkert (D)

Department of Neurobiology, Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.

Michal Linial (M)

Department of Biological Chemistry, Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, Israel.

Esther Oiknine-Djian (E)

Hadassah Medical Center, Jerusalem, Israel.

Michal Mandelboim (M)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.
School of Public Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.

Zvi Livneh (Z)

Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.

Gilat Shenhav-Saltzman (G)

Department of Internal Medicine, Sheba Medical Center, Tel Hashomer, Ramat-Gan, Israel.
Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.

Ella Mendelson (E)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.
School of Public Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.

Dana Wolf (D)

Hadassah Medical Center, Jerusalem, Israel.

Moran Szwarcwort-Cohen (M)

Virology Laboratory, Rambam Health Care Campus, Haifa, Israel.

Orna Mor (O)

Central Virology Laboratory, Ministry of Health, Chaim Sheba Medical Center, Tel-Hashomer, Ramat-Gan, Israel.
School of Public Health, Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.

Yair Lewis (Y)

Maccabi Healthcare Services, Tel Aviv, Israel.

Danny Zeevi (D)

Department of Biotechnology, Hadassah Academic College, Jerusalem, Israel.

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