key: cord-0898385-gfpiyquk
authors: Hirotsu, Y.; Maejima, M.; Shibusawa, M.; Nagakubo, Y.; Hosaka, K.; Amemiya, K.; Sueki, H.; Hayakawa, M.; Mochizuki, H.; Omata, M.
title: Pooling RT-PCR test of SARS-CoV-2 for large cohort of 'healthy' and infection-suspected patients: A prospective and consecutive study on 1,000 individuals
date: 2020-05-06
journal: nan
DOI: 10.1101/2020.05.04.20088146
sha: dcfeebcf556f78c490de4ec0700eb6c36da796e1
doc_id: 898385
cord_uid: gfpiyquk

Background SARS-CoV-2 testing reagents are expected to become in short supply worldwide. However, little is unknown whether the pooling strategy detects SARS-CoV-2 with accuracy. Method To validate the feasibility of pooling samples, serial dilution analysis and spike-in experiment were conducted using synthetic DNA and nucleic acids extracted from SARS-CoV-2 positive and negative patients. Furthermore, we studied a total of 1,000 individuals, who were 667 'healthy' (195 healthcare workers and 472 hospitalized patients with other disorders than COVID-19 infection) individuals and 333 infection-suspected patients with cough and fever, were tested. Results Serial dilution analysis showed the limit of detection of around 10-100 copies according to National Institute of Infectious Diseases, Japan. Spike-in experiment demonstrated RT-qPCR detect positive signal in pooling samples of SARS-CoV-2 negative and positive patient at the 5-, 10-, 20-fold dilution. By screening with pooling strategy by the end of April, 2020, there are 12 COVID-19 patients in 333 infection suspected patients (3.6%) and zero in 667 'healthy'. We obtained these results with total running 538 times (instead of 1,000 times) by pooling strategy. Conclusion Pooling samples is feasible for saving test reagents and detecting SARS-CoV-2 in clinical setting to prevent the spread of the virus and nosocomial transmission.

to save time and reagents under short supply conditions and prevent delays in reporting results. This could provide the prevalence of ongoing infection in 667 healthy in our distinct, west of Tokyo.

We collected nasopharyngeal swabs between March 11 and April 28, 2020 at Yamanashi Central Hospital. All samples were obtained with cotton swab and universal transport media (Copan, Murrieta, CA). To screen whether medical staffs and hospitalized patients were infected with SARS-CoV-2, we tested a total of 1,000 samples from 1,000 individuals. By pooling strategy, we tested a total of 538 samples (445 individuals and 93 pools). One pooled batch was made of 5 to 10 samples. This study was approved by the Institutional Review Board at Yamanashi Central Hospital and complied with Declaration of Helsinki principles.

Total nucleic acid was automatically isolated from nasopharyngeal swabs using . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted May 6, 2020. . CC-BY-NC-ND 4.0 International license It is made available under a is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity.

The copyright holder for this preprint this version posted May 6, 2020. 

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Pooling of urine samples for