Peer Review History

Original SubmissionMay 10, 2020
Decision Letter - Oscar Millet, Editor


Boosting test-efficiency by pooled testing strategies for



Dear Dr. Hanel,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Specifically, one of the reviewers raised a number of issues that need to be addressed before the manuscript becomes acceptable. Of particular significance is the inclusion of COVID-19 related data to support the findings.

Please submit your revised manuscript by Aug 13 2020 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at When you're ready to submit your revision, log on to and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
  • A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
  • An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in to enhance the reproducibility of your results. assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see:

We look forward to receiving your revised manuscript.

Kind regards,

Oscar Millet

Academic Editor


Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at and

2. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. PLOS defines a study's minimal data set as the underlying data used to reach the conclusions drawn in the manuscript and any additional data required to replicate the reported study findings in their entirety. All PLOS journals require that the minimal data set be made fully available. For more information about our data policy, please see

Upon re-submitting your revised manuscript, please upload your study’s minimal underlying data set as either Supporting Information files or to a stable, public repository and include the relevant URLs, DOIs, or accession numbers within your revised cover letter. For a list of acceptable repositories, please see Any potentially identifying patient information must be fully anonymized.

Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access.

We will update your Data Availability statement to reflect the information you provide in your cover letter.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Partly


2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: N/A


3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: No


4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: No


5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: excellent and important practical manuscript. citation which arrives at similar conclusion and provides direct experimental evidence : Lancet Infect Dis 2020

Published Online

April 28, 2020


might bbe cited

A limitation which might be briefly discussed is the differential sensitivity and specificity of specific "PCR" tests which use one or two) target sequences, and there are small differences between viral target genes and the abundance of target RNA species. Efficiency of the specific methods used for RNA extraction and reverse transcription are variables that may effect sensitivity and ultimately limit pooling.

Reviewer #2: 1. Is it reasonable to assume the false positive/negative rate does not change when switched from the regular testing to pooled testing? Does it apply to COVID-19 screening? It is hard to believe using a pool of size 32 or 64 can have the same false rates as testing the individuals one-by-one.

2. What is the choice of r in practice? How does it affect test efficiency? Following the majority rule, suppose r=5, then in practice, once you observed 3 of them were positive, there is no need to test the remaining 2 replicates. How does this affect the calculation of number of tests needed?

3. Line 47 on Page 2, I believe it should be "If the pooled sample is declared positive, we test each individual in the group separately" because of the majority rule you proposed.

4. The derivation of (3-5) in-explicitly assumed that given the true infection statues, the test results are mutually independent. Is this assumption supported by COVID-19 tests? If COVID-19 tests declare a sample as positive if the measured viral loads exceed a predetermined threshold, then this assumption does not hold.

5. There is no COVID-19 data supporting the methodology.

6. Literature review is not sufficient. See Kim et al. (2009, Biometrics).


6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at Please note that Supporting Information files do not need this step.

Revision 1

Dear Editor and Reviewer.

Thank you for you efforts and the points you raised, which we believe has lead to a significant improvement of our manuscript.

A detailed response to the points you raised are included in both the Cover letter and the Response Letter (which up to a heading are identical)

We hope that the improvements of the manuscript now make it acceptable for publication.

Looking forward to hearing from you,

With kind regards

Rudolf Hanel

Submitted filename: Response to Reviewers 14.08.2020.pdf
Decision Letter - Oscar Millet, Editor

Boosting test-efficiency by pooled testing for SARS-CoV-2 --

formula for optimal pool size


Dear Dr. Hanel,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact

Kind regards,

Oscar Millet

Academic Editor


Additional Editor Comments (optional):

Reviewers' comments:

Formally Accepted
Acceptance Letter - Oscar Millet, Editor


Boosting test-efficiency by pooled testing for SARS-CoV-2 – formula for optimal pool size

Dear Dr. Hanel:

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact

If we can help with anything else, please email us at

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Oscar Millet

Academic Editor


Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .