Evaluation of culture- and PCR-based methods for detecting Burkholderia pseudomallei in soil samples in Thailand

Didtawa Suttisak; Charlene Mae Salao Cagape; Rathanin Seng; Natnaree Saiprom; Wongwarut Boonyanugomol; Kamolchanok Rukseree; Paul J. Brett; Mary N. Burtnick; Narisara Chantratita

doi:10.1371/journal.pntd.0013840

Peer Review History

Original SubmissionJuly 8, 2025
25 Aug 2025 Decision Letter - Elsio Wunder Jr, Editor PNTD-D-25-01107 Evaluation of culture- and PCR-based methods for detecting Burkholderia pseudomallei in soil samples in Thailand PLOS Neglected Tropical Diseases Dear Dr. Chantratita, Thank you for submitting your manuscript to PLOS Neglected Tropical Diseases. After careful consideration, we feel that it has merit but does not fully meet PLOS Neglected Tropical Diseases'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. Please pay attention to the comments provided by both reviewers who are the experts in the field of environmental surveys of Burkholderia pseudomallei. Please note that the review report from Reviewer 2 is provided in the attachment. Please submit your revised manuscript within 60 days Oct 24 2025 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 plosntds@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pntd/ 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 editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. This file does not need to include responses to any formatting updates and technical items listed in the 'Journal Requirements' section below. * 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, competing interests statement, or data availability statement, please make these updates within the submission form at the time of resubmission. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. We look forward to receiving your revised manuscript. Kind regards, Apichai Tuanyok, Ph.D. Guest Editor PLOS Neglected Tropical Diseases Elsio Wunder Jr Section Editor PLOS Neglected Tropical Diseases Shaden Kamhawi co-Editor-in-Chief PLOS Neglected Tropical Diseases orcid.org/0000-0003-4304-636XX Paul Brindley co-Editor-in-Chief PLOS Neglected Tropical Diseases orcid.org/0000-0003-1765-0002 Additional Editor Comments (if provided): Journal Requirements: If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. [Note: HTML markup is below. Please do not edit.] Reviewers' Comments: 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: (No Response) Reviewer #2: The global distribution of the soil pathogen B. pseudomallei is unclear. Among other reasons, this is also due to methodological challenges in detecting this pathogen in the environment. This manuscript addresses this important issue by validating previously described culture and PCR methods using spiked soil samples. In addition, culture and PCR methods were used to test the hypothesis that seropositivity in healthy individuals is associated with B. pseudomallei-positive soil samples of the respective households. The study design is appropriate to adress the stated objectives ******** 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: (No Response) Reviewer #2: The results are generally clearly presented. Table and figures are of sufficient quality. ****** 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: (No Response) Reviewer #2: The conclusions are generally justified. However the following point should be clarified and discussed: Presumably, the soil samples for DNA extraction and for the cultures were separate sub-samples of the original 200 g samples? Given that the distribution of B. pseudomallei can vary greatly even within a small volume, the large differences in the soil quantities used for the PCR and culture protocols could at least partly explain the low PCR positivty rate of culture-positive samples (?). ****** 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: (No Response) Reviewer #2: Line 490 Reference not correct? Line 512 to 513 Meaning 20% higher among younger people than the average? Line 556 to 561 The authors only refer to previous studies that detected B. pseudomallei DNA after culture enrichment. However, there are a number of studies which also applied PCR directly to soil extracts. These studies should be cited and briefly discussed. Line 559 ...from soil ( ) Reference should be added. Abstract line 39 to 43 The culture results of samples from seronegative and seropositive households should be provided ****** 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: (No Response) Reviewer #2: (No Response) ****** 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: Yes: Jeffrey Warner 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.] Figure resubmission: Reproducibility:** To enhance the reproducibility of your results, we recommend that authors of applicable studies deposit laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. Additionally, PLOS ONE offers an option to publish peer-reviewed clinical study protocols. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols Attachments Attachment Submitted filename: Suttisak et al review.docx https://doi.org/10.1371/journal.pntd.0013840.r001
Revision 1
23 Oct 2025 Author Response Attachments Attachment Submitted filename: Point by point response for reviewer_Final.docx https://doi.org/10.1371/journal.pntd.0013840.r002
8 Dec 2025 Decision Letter - Elsio Wunder Jr, Editor Dear Dr. Chantratita, We are pleased to inform you that your manuscript 'Evaluation of culture- and PCR-based methods for detecting Burkholderia pseudomallei in soil samples in Thailand' 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, Apichai Tuanyok, Ph.D. Guest Editor PLOS Neglected Tropical Diseases Elsio Wunder Jr Section Editor PLOS Neglected Tropical Diseases Shaden Kamhawi co-Editor-in-Chief PLOS Neglected Tropical Diseases orcid.org/0000-0003-4304-636XX Paul Brindley co-Editor-in-Chief PLOS Neglected Tropical Diseases orcid.org/0000-0003-1765-0002 ********************************************************* p.p1 {margin: 0.0px 0.0px 0.0px 0.0px; line-height: 16.0px; font: 14.0px Arial; color: #323333; -webkit-text-stroke: #323333}span.s1 {font-kerning: none 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: Objectives clear Reviewer #2: (No Response) ****** 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: Results are complete Reviewer #2: (No Response) ****** 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: Good conclusions Reviewer #2: (No Response) ****** 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: (No Response) Reviewer #2: (No Response) ****** 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: (No Response) Reviewer #2: (No Response) ****** 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: Yes:** Jeff Warner Reviewer #2: No https://doi.org/10.1371/journal.pntd.0013840.r003
Formally Accepted
Acceptance Letter - Elsio Wunder Jr, Editor Dear Dr. Chantratita, We are delighted to inform you that your manuscript, "Evaluation of culture- and PCR-based methods for detecting Burkholderia pseudomallei in soil samples in Thailand," has been formally accepted for publication in PLOS Neglected Tropical Diseases. We have now passed your article onto the PLOS Production Department who will complete the rest of the publication process. All authors will receive a confirmation email upon publication. The corresponding author will soon be receiving a typeset proof for review, to ensure errors have not been introduced during production. Please review the PDF proof of your manuscript carefully, as this is the last chance to correct any scientific or type-setting errors. Please note that major changes, or those which affect the scientific understanding of the work, will likely cause delays to the publication date of your manuscript. Note: Proofs for Front Matter articles (Editorial, Viewpoint, Symposium, Review, etc...) are generated on a different schedule and may not be made available as quickly. Soon after your final files are uploaded, the early version of your manuscript will be published online unless you opted out of this process. The date of the early version will be your article's publication date. The final article will be published to the same URL, and all versions of the paper will be accessible to readers. For Research Articles, you will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. Thank you again for supporting open-access publishing; we are looking forward to publishing your work in PLOS Neglected Tropical Diseases. Best regards, Shaden Kamhawi co-Editor-in-Chief PLOS Neglected Tropical Diseases Paul Brindley co-Editor-in-Chief PLOS Neglected Tropical Diseases https://doi.org/10.1371/journal.pntd.0013840.r004

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