Pooling RT-qPCR testing for SARS-CoV-2 in 1000 individuals of healthy and infection-suspected patients View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2020-11-03

AUTHORS

Yosuke Hirotsu, Makoto Maejima, Masahiro Shibusawa, Yuki Nagakubo, Kazuhiro Hosaka, Kenji Amemiya, Hitomi Sueki, Miyoko Hayakawa, Hitoshi Mochizuki, Toshiharu Tsutsui, Yumiko Kakizaki, Yoshihiro Miyashita, Masao Omata

ABSTRACT

Severe acute respiratory coronavirus 2 (SARS-CoV-2) testing reagents are expected to become scarce worldwide. However, little is known regarding whether pooling of samples accurately detects SARS-CoV-2. To validate the feasibility of pooling samples, serial dilution analysis and spike-in experiments were conducted using synthetic DNA and nucleic acids extracted from SARS-CoV-2-positive and -negative patients. Furthermore, we studied 1000 individuals, 667 of whom were "healthy" individuals (195 healthcare workers and 472 hospitalized patients with disorders other than COVID-19 infection), and 333 infection-suspected patients with cough and fever. Serial dilution analysis showed a limit of detection of around 10-100 viral genome copies according to the protocol of the National Institute of Infectious Diseases, Japan. Spike-in experiments demonstrated that RT-qPCR detected positive signals in pooled samples with SARS-CoV-2-negative and -positive patients at 5-, 10-, 20-fold dilutions. By screening with this pooling strategy, by the end of April 2020 there were 12 SARS-CoV-2-positive patients in 333 infection-suspected patients (3.6%) and zero in 667 "healthy" controls. We obtained these results with a total of 538 runs using the pooling strategy, compared with 1000 standard runs. In a prospective study, we successfully detected SARS-CoV-2 using 10- to 20-fold diluted samples of nasopharyngeal swabs from eighteen COVID-19 patients with wide ranges of viral load. Pooling sample is feasible for conserving test reagents and detecting SARS-CoV-2 in clinical settings. This strategy will help us to research the prevalence infected individuals and provide infected-status information to prevent the spread of the virus and nosocomial transmission. More... »

PAGES

18899

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41598-020-76043-z

DOI

http://dx.doi.org/10.1038/s41598-020-76043-z

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https://app.dimensions.ai/details/publication/pub.1132260155

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/33144632


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69 pooling
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73 positive signals
74 prevalence
75 prospective study
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77 range
78 reagents
79 respiratory coronavirus 2 (SARS-CoV-2) testing reagents
80 results
81 run
82 samples
83 serial dilution analysis
84 setting
85 signals
86 spikes
87 spread
88 standard run
89 strategies
90 study
91 swabs
92 synthetic DNA
93 test reagents
94 testing
95 testing reagents
96 total
97 transmission
98 viral genome copies
99 viral load
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