PONE-D-23-37922One-instrument, objective microsatellite instability analysis using high-resolution meltPLOS ONE

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Academic Editor

PLOS ONE

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Additional Editor Comments:

This manuscript was carefully reviewed by 2 experts. Both of them highly evaluated this study, but there are several minor issues which need to be addressed before acceptance. Please respond to each of the reviewer comments.

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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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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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: Yes

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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: Yes

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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: 1. I thought the author's point in this paper was that MicroSight® MSI 1-Step HRM Analysis, which can evaluate MSI without fragment analysis, could be as good as MSI testing using traditional fragment analysis. This point is not clear in this paper, and I would recommend that the paper be revised. Similarly, the authors should present the results of the investigation of whether there is a difference in the instability of microsatellite markers (BAT-25, BAT-26, NR-22, NR-24, MONO27) between conventional fragment analysis methods and MicroSight® MSI 1-Step HRM Analysis. It would be trivial to compare MicroSight® MSI 1-Step HRM Analysis with other MSI testing or to investigate whether differences in DNA extraction methods affect the MicroSight® MSI 1-Step HRM Analysis.

2. Many reports have evaluated microsatellite instability using BAT-25, BAT-26, NR-21, NR-24, and NR-27 (e.g., Ann Oncol. 2019 Aug 1;30(8):1232-1243. doi: 10.1093). Please describe why the authors used NR-22 instead of NR-21.

Reviewer #2: The manuscript entitled "One-instrument, objective microsatellite instability analysis using high-resolution melt" highlighted that the MicroSight® MSI assay showed a high repeatability regardless of DNA extraction method and PCR platform, and a 100% agreement of the MSI status with the PCR fragment analysis method.

- The AUthors should provide the expand forms for all acronyms, including gene acronyms, through the text when they first appear.

- Gene acronyms should be written in italics.

- The Authors should consider to cite the role of pre-analitics in the discussion (PMID: 34440647, PMID: 32887373)

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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

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One-instrument, objective microsatellite instability analysis using high-resolution melt

PONE-D-23-37922R1

Dear Dr. Petersen,

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.

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Kind regards,

Hiromu Suzuki, M.D., Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The authors addressed all issues raised by the reviewer.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. 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: Yes

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

Reviewer #1: Yes

Reviewer #2: Yes

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4. 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: Yes

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5. 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: Yes

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6. 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: (No Response)

Reviewer #2: The authors have addressed all my concerns and I have no further comments.

The authors have addressed all my concerns and I have no further comments.

The authors have addressed all my concerns and I have no further comments.

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7. 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

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