Dear Dr. Blacksell,

Thank you very much for submitting your manuscript "Diagnostic accuracy of DPP® Fever Panel II Asia tests for tropical fever diagnosis" for consideration at PLOS Neglected Tropical Diseases. As with all papers reviewed by the journal, your manuscript was reviewed by members of the editorial board and by several independent reviewers. The reviewers appreciated the attention to an important topic. Based on the reviews, we are likely to accept this manuscript for publication, providing that you modify the manuscript according to the review recommendations.

Please prepare and submit your revised manuscript within 30 days. If you anticipate any delay, please let us know the expected resubmission date by replying to this email.

Please include a paragraph about limitations of the assay in the discussion based on reviewer 1 comments.

When you are ready to resubmit, please upload the following:

[1] A letter containing a detailed list of your responses to all review comments, and a description of the changes you have made in the manuscript.

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[2] Two versions of the revised manuscript: one with either highlights or tracked changes denoting where the text has been changed; the other a clean version (uploaded as the manuscript file).

Important additional instructions are given below your reviewer comments.

Thank you again for your submission to our journal. We hope that our editorial process has been constructive so far, and we welcome your feedback at any time. Please don't hesitate to contact us if you have any questions or comments.

Sincerely,

Thomas C Darton

Academic Editor

PLOS Neglected Tropical Diseases

Shaden Kamhawi

Editor-in-Chief

PLOS Neglected Tropical Diseases

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Reviewer's Responses to Questions

Key Review Criteria Required for Acceptance?

As you describe the new analyses required for acceptance, please consider the following:

Methods

-Are the objectives of the study clearly articulated with a clear testable hypothesis stated?

-Is the study design appropriate to address the stated objectives?

-Is the population clearly described and appropriate for the hypothesis being tested?

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested?

-Were correct statistical analysis used to support conclusions?

-Are there concerns about ethical or regulatory requirements being met?

Reviewer #1: The authors should state how the samples used as "true positives" were confirmed to be so. Was it based on clinical presentation, some other diagnostic test or a combination of both ? When developing and testing a new assay it is crucial that the "known positives" are indeed "genuine true positives".

Reviewer #2: -Are the objectives of the study clearly articulated with a clear testable hypothesis stated? - Yes

-Is the study design appropriate to address the stated objectives? - Yes

-Is the population clearly described and appropriate for the hypothesis being tested? - Yes

-Is the sample size sufficient to ensure adequate power to address the hypothesis being tested? - Authors addressed in limitation

-Were correct statistical analysis used to support conclusions? - Yes

-Are there concerns about ethical or regulatory requirements being met? - Yes

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Results

-Does the analysis presented match the analysis plan?

-Are the results clearly and completely presented?

-Are the figures (Tables, Images) of sufficient quality for clarity?

Reviewer #1: Yes

What is the "CPS" antigen of B.pseudomallei ? Some readers (including me ) may not know what it is !

Reviewer #2: -Does the analysis presented match the analysis plan? - Suggested for Minor revision

-Are the results clearly and completely presented? - Yes

-Are the figures (Tables, Images) of sufficient quality for clarity? - Yes

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Conclusions

-Are the conclusions supported by the data presented?

-Are the limitations of analysis clearly described?

-Do the authors discuss how these data can be helpful to advance our understanding of the topic under study?

-Is public health relevance addressed?

Reviewer #1: This new assay is not all that impressive and I think this needs to be stated in the Conclusions. Its a bit misleading to just say "it is comparable to that of commonly used RDTs", although admittedly these aren't that good either !

Reviewer #2: Suggested for Minor revision

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Editorial and Data Presentation Modifications?

Use this section for editorial suggestions as well as relatively minor modifications of existing data that would enhance clarity. If the only modifications needed are minor and/or editorial, you may wish to recommend “Minor Revision” or “Accept”.

Reviewer #1: The comparison between the 2 readers is not that interesting to the general reader, although it may well be so to the sponsoring company. This could be summarised in the paper rather than gone into in detail.

"WB" as an abbreviation usually means "Western Blot", not "whole blood". Was it really "whole blood" or was it plasma ? Could another abbreviation be used instead of "WB" ?

Reviewer #2: (No Response)

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Summary and General Comments

Use this section to provide overall comments, discuss strengths/weaknesses of the study, novelty, significance, general execution and scholarship. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. If requesting major revision, please articulate the new experiments that are needed.

Reviewer #1: Interesting paper but apart from a few assays its not that impressive with only modest sensitivities and specificities for most of the tropical infections tested for.

line 103. "lack of non-specificity". Don't you mean "lack of specificity" ?

line 144 "will be evaluated" should be "was evaluated"

line 342 "...from severe disease if validated with further study" reads better.

line 360 "igG develops latently" ? Maybe "IgG develops slowly".

line 379 suggest change to "hospitalisation and this early sampling may have contributed to lower sensitivity and specificity for detecting antibody levels."

lines 387-8 suggest change to "Repeating the assay after a period sufficient to allow for seroconversion is recommended to provide greater confidence in the result."

line 414. What is "FIND" ?

line 649. change "compiled" to "combined"

Reviewer #2: The screening and diagnosis of many infectious syndromes using a single sample at the time of a patient visit to a hospital or at the time of a field visit by healthcare workers in the community is very important for timely diagnosis and starting appropriate therapy without delay. The study focuses on this aspect in febrile illness patients with multiplexing the point-of-care tests. I have added a few comments for revision for further comments or recommendations.

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Reviewer #1: No

Reviewer #2: Yes: Sivanantham Krishnamoorthi

Figure Files:

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References

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article's retracted status in the References list and also include a citation and full reference for the retraction notice.

Attachments

Attachment

Submitted filename: PNTD-D-23-00978 Reviewer Comment.docx

Attachment

Submitted filename: PNTD-D-23-00978_reviewer (1).pdf

Dear Dr. Blacksell,

We are pleased to inform you that your manuscript 'Diagnostic accuracy of DPP® Fever Panel II Asia tests for tropical fever diagnosis Subtitle: Validation of DPP® Fever Panel II' has been provisionally accepted for publication in PLOS Neglected Tropical Diseases.

Before your manuscript can be formally accepted you will need to complete some formatting changes, which you will receive in a follow up email. A member of our team will be in touch with a set of requests.

Please note that your manuscript will not be scheduled for publication until you have made the required changes, so a swift response is appreciated.

IMPORTANT: The editorial review process is now complete. PLOS will only permit corrections to spelling, formatting or significant scientific errors from this point onwards. Requests for major changes, or any which affect the scientific understanding of your work, will cause delays to the publication date of your manuscript.

Should you, your institution's press office or the journal office choose to press release your paper, you will automatically be opted out of early publication. We ask that you notify us now if you or your institution is planning to press release the article. All press must be co-ordinated with PLOS.

Thank you again for supporting Open Access publishing; we are looking forward to publishing your work in PLOS Neglected Tropical Diseases.

Best regards,

Thomas C Darton

Academic Editor

PLOS Neglected Tropical Diseases

Shaden Kamhawi

Editor-in-Chief

PLOS Neglected Tropical Diseases

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Many thanks for addressing the reviewer comments raised, which has been done in a clear and thorough way.