Multiplex detection of “Candidatus Liberibacter asiaticus” and Spiroplasma citri by qPCR and droplet digital PCR

Yogita Maheshwari; Vijayanandraj Selvaraj; Kristine Godfrey; Subhas Hajeri; Raymond Yokomi

doi:10.1371/journal.pone.0242392

Peer Review History

Original SubmissionOctober 30, 2020
17 Feb 2021 Decision Letter - Zonghua Wang, Editor PONE-D-20-34072 Multiplex detection of “Candidatus Liberibacter asiaticus” and Spiroplasma citri by qPCR and droplet digital PCR PLOS ONE Dear Dr. Yokomi, 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. Please submit your revised manuscript by Apr 03 2021 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 plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 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: The manuscript presented by Maheshwari et al presents the multiplex detection of “Candidatus Liberibacter asiaticus ” and Spiroplasma citri by qPCR and droplet digital PCR. This study contains major results that deserve to be published and made available to the scientific community. The writing needs to be better focused on the main objectives instead of being descriptive. In discussion section, one additional paragraph could be added to explain any new parameter or technical improvement by comparison with previous researches, as a number of papers reporting sensitive detection techniques for Clas and S. citri. Reviewer #2: I think that qPCR and droplet digital are good methods for Candidatus Liberibacter asiaticus and Spiroplasma citri detection. If they were used to detect in the field, do you compare the cost of droplet and qPCR? Which one is more economic? The primer of COXf was lost 3 bases as the reference, does the amplification efficiency is the same? ****** 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? 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Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0242392.r001
Revision 1
25 Feb 2021 Author Response Reviewer 1 • The writing needs to be better focused on the main objectives instead of being descriptive. The research objective is stated more clearly in Abstract Page 2, Lines 29-31. To improve focus, clarity was added in the Discussion Page 13, Lines 270-271; Lines 279-282; Page 14, Lines 287-295; Lines 298-302. The major data presented, however, needs to stay as written as documentation for the efficacy and reliability of the assays which are needed if the procedures are to be used for regulatory samples. • In discussion section, one additional paragraph could be added to explain any new parameter or technical improvement by comparison with previous researches, as a number of papers reporting sensitive detection techniques for Clas and S. citri. Although there are numerous non- PCR-based assays developed for CLas and S. citri detection, the regulatory standard is still qPCR. ddPCR was added to serve as secondary test for more precise pathogen detection, albeit, more costly and less high throughput. Some text was added in Discussion, Page 14, Lines 293-295; Lines 298-302. Reviewer 2 • If they were used to detect in the field, do you compare the cost of droplet and qPCR? Which one is more economic? Comparative costs and time to complete ddPCR versus qPCR was added in discussion, Page 14, Lines 287-292. • The primer of COXf was lost 3 bases as the reference, does the amplification efficiency is the same? Thank you for pointing out this disparity. It was a typo and page 6 Table 1 has been corrected. Attachments Attachment Submitted filename: Response to Reviewers.pdf https://doi.org/10.1371/journal.pone.0242392.r002
3 Mar 2021 Decision Letter - Zonghua Wang, Editor Multiplex detection of “Candidatus Liberibacter asiaticus” and Spiroplasma citri by qPCR and droplet digital PCR PONE-D-20-34072R1 Dear Dr. Yokomi, 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 http://www.editorialmanager.com/pone/, 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 authorbilling@plos.org. 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 onepress@plos.org. Kind regards, Zonghua Wang, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0242392.r003
Formally Accepted
9 Mar 2021 Acceptance Letter - Zonghua Wang, Editor PONE-D-20-34072R1 Multiplex detection of “Candidatus Liberibacter asiaticus” and Spiroplasma citri by qPCR and droplet digital PCR Dear Dr. Yokomi: 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 onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Prof. Zonghua Wang Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0242392.r004

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