PONE-D-20-23819

Borrelia burgdorferi and Borrelia miyamotoi Seroprevalence in California Blood Donors

PLOS ONE

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"The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Dr. Bloch is a member of the U.S. Food and Drug Administration (FDA) Blood Products Advisory Committee. Any views or opinions expressed in this manuscript are Dr. Bloch's and are based on his own scientific expertise and professional judgment; they do not necessarily represent the views of the Blood Products Advisory Committee or the formal position of the FDA and also do not bind or otherwise obligate or commit either the Advisory Committee or the FDA to the views expressed. The views and opinions expressed herein are those of the authors alone and do not represent the official position of the Centers for Disease Control and Prevention."

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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: I Don't Know

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: Review: Borrelia burgdorferi and Borrelia miyamotoi Seroprevalence in California Blood Donors.

This study assessed the seroprevalence of B. burgdorferi and B. miyamotoi in 1700 blood donors in California. Donors were selected from counties with reported high prevalence of B. burgdorferi infection as well as those with low prevalence. Of the 1700 donors, 91 were positive (or equivocal) by a standalone C6 ELISA assay for B. burgdorferi. When two tier (STTT or MTTT) assay was applied only 8 of the 91 donors were positive for B. burgdorferi antibodies. Two donors were positive for B. miyamotoi antibodies. No major differences in seroprevalence were observed between high and low prevalence areas. In total 0.47% of patients had serological evidence of B. burgdorferi infection and 0.12% had serological evidence of B. miyamotoi infection. The findings demonstrate low prevalence of B. burgdorferi and B. miyamotoi infection in California.

A key strength of this study is the assessment of seroprevalence for both agents at two independent testing sites (UCD and CDC) and the use of 2 testing methodologies (STTT and MTTT) for B. burgdorferi. The results are largely consistent with previous findings. The major comment is regarding the data presentation in the Results, which could be improved.

1) Figure 1 should include the number of samples that came from each county as well as the percentage. Figure Legends should be provided.

2) The 2nd paragraph of the Results (C6 ELISA Screen) provides information on how the 91 patients were selected for downstream analyses, but this information is not presented anywhere. My suggestion is to present this in a form of a Table or a flow chart since all subsequent analyses in the manuscript stem from this selection/screen.

3) In the second paragraph of the C6 ELISA Screen section, the authors discuss the distribution of Hispanics and non-Hispanics among low and high prevalence areas in California. They make a point that individuals with positive / equivocal result were usually non-Hispanic however one would assume this based on the lower prevalence of Hispanics in this cohort. The seroprevalence to C6 peptide should be assessed directly in Hispanics vs non-Hispanics taking into account the sample size for each.

4) The authors selected randomly 9 negative samples by standalone C6 ELISA, two thirds of which were female. What is the rationale for selecting 9 samples (vs more) and why were they not gender matched with the C6 seropositive populations?

5) Table 2 should include a row that lists the C6 findings for all 1700 patients. It should also include data for all 941 samples from high endemic area, as well as for 759 patients from low enedemic area; followed by the data for each county in that area. The data need to be clearly labeled (what are the data in parenthesis? Do the data represent positive results?). It would help to present the number of positives in addition to percentages.

6) The paragraph on B. miyamotoi GlpQ seroreactivity discusses seroprevalence in high vs low endemic regions for B. burgdorferi. This discussion is based on only two positive samples, and it is difficult to conclude much in the difference in prevalence between the two regions, except that the seroprevalence is relatively low. One could also postulate that the infection could have been acquired elsewhere and not in the counties/state where the samples were obtained.

7) A similar comment holds true for the paragraph on B. burgdorferi IgG western blot. The authors provide percentages and CI for seroprevalence in Marin county, but only one donor was seropositive there. With such small numbers it is difficult to assess the significance of seroprevalence and one should be careful to not overinterpret the data. For example, is it fair to state that San Luis Obispo county had the highest prevalence in the low endemic area since it had the only positive case (out of 759) in this region?

8) Line 369 in the Discussion: is it low specificity or high sensitivity of C6 ELISA that would over-estimate true exposure?

Reviewer #2: This is a very well written paper from several experienced investigators. They report a comprehensive sero-survey of B. burgdorferi and B. miyamotoi in counties of high and low endemicity for LD in California. The study design is straightforward and the results confirm previous observations of low prevalence of LD in CA (<0.5%). They found an even lower prevalence of antibodies to B. miyamotoi (0.1%). It is interesting that they found that the two positive miyamotoi cases were positive in C6 and Qlp. The following paper should be discussed: Borrelia miyamotoi infection leads to cross-reactive antibodies to the C6 peptide in mice and men. Koetsveld J, Platonov AE, Kuleshov K, Wagemakers A, Hoornstra D, Ang W, Szekeres S, van Duijvendijk GLA, Fikrig E, Embers ME, Sprong H, Hovius JW. Clin Microbiol Infect. 2020 Apr;26(4):513.e1-513.e6. doi: 10.1016/j.cmi.2019.07.026. Epub 2019 Aug 9. PMID: 31404672

The results are well discussed and limitations of the study have be identified. I have no further major or minor suggestions to improve this paper.

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Reviewer #1: No

Reviewer #2: No

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Attachments

Attachment

Submitted filename: PlosOne 2020 Brummitt etal.doc

Borrelia burgdorferi and Borrelia miyamotoi Seroprevalence in California Blood Donors

PONE-D-20-23819R1

Dear Mrs. Brummitt,

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.

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Kind regards,

R. Mark Wooten, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

**********

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: The revised manuscript addresses the reviewer’s concerns. It brings to light important issue regarding seroprevalence of B. burgdorferi and B. miyamotoi in California which caries public health implications.

Reviewer #2: (No Response)

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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