Therapeutic efficacy of Chloroquine for the treatment of uncomplicated Plasmodium vivax infection in Shewa Robit, Northeast Ethiopia

Habtamu Belay; Megbaru Alemu; Tadesse Hailu; Hussein Mohammed; Heven Sime; Henok Hailegeorgies; Bokretsion Gidey; Mebrahtom Haile; Gudissa Assefa; Worku Bekele; Mihreteab Alebachew Reta; Andargachew Almaw Tamene; Geremew Tasew; Ashenafi Assefa

doi:10.1371/journal.pone.0277362

Peer Review History

Original SubmissionJune 26, 2022
26 Jul 2022 Decision Letter - José Luiz Fernandes Vieira, Editor PONE-D-22-18161Therapeutic efficacy of Chloroquine for the treatment of uncomplicated Plasmodium vivax infection in Shewa Robit, Northeast EthiopiaPLOS ONE Dear Dr. Temesgne 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. ACADEMIC EDITOR: The maniscript is interesting, but several issue related to the methodology section, inclusion and exclusion creiteria and sample effort should be adressed in the manuscript. Please follow the reccomendations of the reviewers. Please submit your revised manuscript by August 28. 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. 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If your ethics statement is written in any section besides the Methods, please move it to the Methods section and delete it from any other section. Please ensure that your ethics statement is included in your manuscript, as the ethics statement entered into the online submission form will not be published alongside your manuscript. [Note: HTML markup is below. Please do not edit.] 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: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: I Don't Know ****** 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 ****** 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 ****** 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: Dear, Congratulations on the study, it has great scientific relevance. Therefore, some questions need to be answered: When analyzing your study, both the abstract and the introduction are contemplating the necessary information; The methodology needs to be further explored. It is still not entirely clear what the inclusion and exclusion criteria for the recruited patients were. In addition, the sample number seems small to me. Wouldn't it have a chance to increase? Another important factor is that no analysis of the biochemical parameters of the patients was performed, these data would give you significant answers to your findings. Why wasn't it done? The results are well described, but at the same time I noticed that the patients did not present itching. Typically, patients who use CQ have these reactions. Was it not identified or did these patients have and/or abandoned the segment? Furthermore, I suggest that in figures 3 and 4 you put the corresponding values in each highlight point for a better understanding for the reader; The discussion presented includes the necessary information. However, by addressing minor limitations you could emphasize that the process of genetic polymorphism could account for drug resistance. This is constantly being discussed in several scientific articles in different locations worldwide. It is also necessary for you to present more accurate information to justify your hypotheses and/or findings; The conclusion is ok. Also, what makes your work different? I hope that this information has been essential for a better understanding of the study. Thank you very much in advance. Reviewer #2: Background 1. Put epidemiology data latest 2. When did the first case of resistance appear in Ethiopia? Insert this information 3. How was resistance studies carried out previously carried out? Methods 4. describe what were the inclusion criteria 5. describe what were the exclusion criteria? 6 describe what were the severe malaria exclusion criteria? 7. Where did this frequency of therapeutic failure come from? 8. What is the expected minimum and maximum percentage for sample calculation? 9. what number of malaria in the studied place for sample calculation? need your population number. 10. what definition of recurrence and clearance of parasitemia? 11. Drug quality analysis was performed? Enter chloroquine information? 12. Did all patients take the same batch of chloroquine? 13. Was it quality control carried out on this batch? 14. Why were these follow-up visits selected? if the patient had a return of malaria on other days or after D28 was it lost? 15. How was treatment on different days describe dosage on each day? 16. Were recorded Baseline data on socio-demographic and clinical characteristics? 17. Has quality assurance been performed for microscopy? 18. What were the criteria for establishing adverse effects? 19. How was security assessed? what criteria? 20. Primary Outcomes and secondary? Define each item of this study. 21. Was it not evaluated clearance and presence of gametocytes? Results 22. Make a flowchart describing how the selection was made and the number of participants included. 23. Were participants who did not take CQ as treatment included? 24. This phrase "couldn't complete the 28-day follow-up and were therefore excluded", needs to be included in the flowchart. 25. In figure 2, insert the correlation confidence interval? p value? 26. what purpose of making correlation? 27. Why was the dosage of chloroquine not performed for classification of recurrence? 28. Why were these data (K-M analysis) not presented through the survival curve? 29. what was the parasitemia of the recurrent case? Put in the figure 3. 30. This study cannot speak about resistance to CQ, as resistance was not confirmed in these participants. 31. What was done to confirm resistance? Discussion 32. Relapse, malabsorption of drug, poor drug quality and re-infection could be contributing factors to treatment failure of P. vivax? Insert into the discussion on this aspect. 33. drug quality control was not performed? 34. Better describe all limitations: sample size, not having performed chloroquine dosage, not performing drug quality control ****** 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? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No 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.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: renamed_058a5.pdf https://doi.org/10.1371/journal.pone.0277362.r001
Revision 1
16 Sep 2022 Author Response Response to reviewers 1. Point by point response to reviewer #1 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 ________________________________________ 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes ________________________________________ 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 ________________________________________ 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 5.Review Comments to the Author Reviewer #1: Dear, Congratulations on the study, it has great scientific relevance. Therefore, some questions need to be answered: When analyzing your study, both the abstract and the introduction are contemplating the necessary information; The methodology needs to be further explored. It is still not entirely clear what the inclusion and exclusion criteria for the recruited patients were. In addition, the sample number seems small to me. Wouldn't it have a chance to increase? Another important factor is that no analysis of the biochemical parameters of the patients was performed, these data would give you significant answers to your findings. Why wasn't it done? The results are well described, but at the same time I noticed that the patients did not present itching. Typically, patients who use CQ have these reactions. Was it not identified or did these patients have and/or abandoned the segment? Furthermore, I suggest that in figures 3 and 4 you put the corresponding values in each highlight point for a better understanding for the reader; The discussion presented includes the necessary information. However, by addressing minor limitations you could emphasize that the process of genetic polymorphism could account for drug resistance. This is constantly being discussed in several scientific articles in different locations worldwide. It is also necessary for you to present more accurate information to justify your hypotheses and/or findings; The conclusion is ok. Also, what makes your work different? I hope that this information has been essential for a better understanding of the study. Thank you very much in advance. Response: Thank you for your constructive comments, we accepted all your suggestions and tried to correct them accordingly. 1. We have followed strictly the WHO guideline “Methods for the surveillance of antimalarial drug efficacy (2009)” accordingly we have followed strict inclusion and exclusion criteria as well as sample size calculation and corrected accordingly in the revised manuscript. The 73-size is the minimum required for such a study. 2. We strongly agree that additional biomedical analysis may strengthen the study however due to strictly following the guideline and limited resources, we were not able to do the biomedical analysis. 3. Itching can be manifested in patients taking CQ. Unfortunately, we observed participants physically and didn’t see any itchy behavior. The participants may not have noticed and didn’t explain it to us. 4. Thank you, comments on figure 3 & 4 corrected accordingly on the revised manuscript. 5. We agree that genetic polymorphism may contribute to drug resistance however, it was beyond the scope of this study, and we have indicated it as a limitation thank you for raising it. 6. Our study may not be totally unique but we produce evidence from a common location and the information is relevant for local and global malaria control efforts. Point by point response to reviewer#2 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 #2: Partly ________________________________________ 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: I Don't Know ________________________________________ 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 #2: Yes ________________________________________ 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 #2: Yes ________________________________________ 5. Review Comments to the Author Reviewer#2: Background 1. Put epidemiology data latest Response: thank you for your suggestion. We have updated the epidemiology data. 2. When did the first case of resistance appear in Ethiopia? Insert this informatioitn Response: thank you, we have corrected accordingly. 3. How was resistance studies carried out previously carried out? Response: Thank you, WHO recommends routine monitoring of drug resistance using its guideline every other year for two to six sites based on country size. There have been continuous but inconsistent studies of malaria drug resistance for the past three decades in Ethiopia. Methods 4. describe what were the inclusion criteria Response: Thank you, corrected accordingly. 5. describe what were the exclusion criteria? Response: Thank you and accepted. 6 describe what were the severe malaria exclusion criteria? Response: Thank you. The severe malaria exclusion criteria are described in the WHO study protocol that we followed. We want to minimize the size of the MS and keep it as a reference. 7. Where did this frequency of therapeutic failure come from? Response: thank you for your question, the frequency of failure was obtained from a previously published study. 8. What is the expected minimum and maximum percentage for sample calculation? Response: Thank you. According to WHO guidelines, in the TES study, the sample size is characterized and calculated to be the minimum number of individuals required for an expected 5% failure is 73, and a maximum of >88 patients. 9. what number of malaria in the studied place for sample calculation? need your population number. Response: Thank you. The population may be important but all self-presenting patients suspected of malaria were diagnosed and if P.vivax infection is confirmed with the parasitemia load, were enrolled in the study team. The study population is the catchment area for Shewa Robit HC, it is estimated to be 60,234. 10. what definition of recurrence and clearance of parasitemia? Response: we have appreciated your question, but the terms clearance and recurrence are well stated in the study protocol that clearance is parasitemia disappearing from the patient sample when examined by microscopy. Recurrence is the reappearance of the parasite in the patient’s blood during the follow-up period. 11. Drug quality analysis was performed? Enter chloroquine information? Response: Thank you. The quality of the study drug was checked and provided by WHO. Information (drugs’ name, manufacturer, lot number, and expired date) are described in the manuscript. No additional quality assessment has been made. 12. Did all patients take the same batch of chloroquine? Response: Thank you. All drugs were in one batch. 13.Was it quality control carried out on this batch? Response: Thank you. The quality concern was done by WHO, as drug provided by. 14. Why were these follow-up visits selected? if the patient had a return of malaria on other days or after D28 was it lost? Response: Thank you, follow-up visits were scheduled based on WHO therapeutic efficacy study protocol and all participants were encouraged to come to the health center if they fill any symptoms in non-scheduled day, therefore, they were not lost. 15. How was treatment on different days describing dosage on each day? Response: Thank you and corrected accordingly. 16. Were recorded Baseline data on socio-demographic and clinical characteristics? Response: Thank you and corrected. The baseline data were recorded on the case screening form and a summary is presented in the manuscript. 17. Has quality assurance been performed for microscopy? Response: Thank you, yes. Experienced microscopists with refresher training participated in the study, the microscopy results were re-read by WHO certified microscopy experts and discrepancies were judged by a third reader. 18. What were the criteria for establishing adverse effects? Response: Thank you, your comment is well appreciated. Adverse effects are established by WHO and clearly stated in the study protocol, and we defined the adverse events in the revised manuscript. Your comment is well appreciated. 19. How was security assessed? What criteria? Response: Thank you. If I got you mean by security assessed? The study site was selected as the sentinel site by the selection criteria of therapeutic study sites previously. This is a regular follow-up study for monitoring the frontline drugs in the country. 20. Primary Outcomes and secondary? Define each item of this study. Response: Thank you, your suggestion is well appreciated, primary outcome can be described in different studies as the cure rate of the drug, and secondary outcomes can be defined as the clearance of fever and parasitemia. But we followed the WHO standard therapeutic study protocol and mentioned the response of the study drug one by one as cure rate, parasite, and fever clearance. 21. Was it not evaluated clearance and presence of gametocytes? Response: Thank you, The parasite clearance and presence of gametocytes were evaluated and presented in Figure 3. Results 22. Make a flowchart describing how the selection was made and the number of participants included. Response: thank you for your suggestion. We thought the total screened, enrolled, and excluded participants are well described on the flowchart (Figure 1). 23. Were participants who did not take CQ as treatment included? Response: Thank you. No, we enrolled participants who were diagnosed as P.vivax infected and treated by only CQ for follow-up. 24. This phrase "couldn't complete the 28-day follow-up and were therefore excluded", needs to be included in the flowchart. Response: The reason to exclude from the study is developing hypersensitivity to the drug. this comment is rephrased and appreciated on the revised manuscript and, on the flow chart stated as “hypersensitive with drug”. 25. In figure 2, insert the correlation confidence interval? p value? Response: thank you and accepted. 26. What purpose of making correlation? Response: Thank you for your question. Making correlations to know the relation between the age of participant and parasitemia, and measure the strength of the linear relationship. 27. Why was the dosage of chloroquine not performed for classification of recurrence? Response: Thank you, dosage of treatment was according to national treatment guidelines. Doing the drug blood level is one of the important things to classify the true treatment failure, but even though only had one treatment failure we didn’t perform the test due to lack of resources and we make it a limitation. 28. Why were these data (K-M analysis) not presented through the survival curve? Response: Thank you, we have made the survival curve but, the curve do not show considerable information due to only one treatment failure on day 28, as explained in table form. 29. what was the parasitemia of the recurrent case? Put in the figure 3. Response: Thank you for your suggestion, the parasitemia of the recurrent case is 1760/µ and included in figure 3 as suggestion. 30. This study cannot speak about resistance to CQ, as resistance was not confirmed in these participants. Response: Thank you, yes this study reported the resistance monitoring activity results in the study area. 31. What was done to confirm resistance? Response: Thank you, we have reported clinical failure resistance was not confirmed due to unable to measure the drug’s blood level stated as a limitation. Discussion 32. Relapse, malabsorption of drug, poor drug quality and re-infection could be contributing factors to treatment failure of P. vivax? Insert into the discussion on this aspect. Response: thank you, we have appreciated your suggestion, and corrected it accordingly. 33. drug quality control was not performed? Response: Thank you, The study drug was provided by the world health organization (WHO) as quality checked with responsibility and we have not been mandated to do drug quality. 34. Better describe all limitations: sample size, not having performed chloroquine dosage, not performing drug quality control Response: Thank you. Thank you, comments are accommodated where appropriate. https://doi.org/10.1371/journal.pone.0277362.r002
14 Oct 2022 Decision Letter - José Luiz Fernandes Vieira, Editor PONE-D-22-18161R1Therapeutic efficacy of Chloroquine for the treatment of uncomplicated Plasmodium vivax infection in Shewa Robit, Northeast EthiopiaPLOS ONE Dear Dr Temesgen, 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 few points raised during the review process. Some few points of the reviewer 2 are necessary, including the exclusion and inclusion criteria; the method used to calculate the previous recurrence, the dose administered to patients was the same or was adjusted by body weight? Please submit your revised manuscript by Nov 28 2022 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. 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, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols. We look forward to receiving your revised manuscript. Kind regards, José Luiz Fernandes Vieira Academic Editor PLOS ONE Journal Requirements: 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. [Note: HTML markup is below. Please do not edit.] 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: (No Response) ******** 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: Partly ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: No ****** 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: No ****** 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: Dear This study made the appropriate corrections recommended during the correction process. Therefore, I suggest accepting the study. Reviewer #2: 1. Inclusion criteria not clear: needs to be made clearer, especially like other WHO criteria 2. Exclusion criteria: needs to be made clearer, especially like other WHO criter 3. Make it clear that previous recurrence calculation 4. State that all participants take the same treatment dose 5. How safe was the treatment? What assessment for this security? 6. Primary and secondary definition. 7. Why were these data (K-M analysis) not presented through the survival curve? The survival curve serves exactly to see the recurrence time because in the table it is not clear. Table 3 is not adequate. ****** 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 ******** [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.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0277362.r003
Revision 2
24 Oct 2022 Author Response Response to reviewers Point-by-point response to reviewer #2 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: (No Response) ________________________________________ 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: Partly ________________________________________ 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: No ________________________________________ 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: No ________________________________________ 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: Dear This study made the appropriate corrections recommended during the correction process. Therefore, I suggest accepting the study. Response: Thank you Reviewer #2: 1. Inclusion criteria not clear: needs to be made clearer, especially like other WHO criteria Response: thank you, we have corrected it accordingly. 2. Exclusion criteria: needs to be made clearer, especially like other WHO criter Response: thank you, corrected accordingly. 3. Make it clear that previous recurrence calculation Response: thank you for your suggestion, we didn’t do a recurrence determination (recrudescence vs reinfection) that we tried to describe as a limitation of the study. 4. State that all participants take the same treatment dose Response: thank you, the dose administered to the study patients was adjusted by body weight, which we made clear on the MS. 5. How safe was the treatment? What assessment for this security? Response: thank you, CQ is a drug used on the national P.vivax treatment guideline and the drug was provided by WHO for the study, no need of doing additional analysis. On the follow-up visit, we assessed the adverse events by direct questioning and using adverse event recording formats. 6. Primary and secondary definition. Response: thank you, primary and secondary treatment outcomes are defined based on WHO treatment outcome definition and we defined the outcomes in the MS on the study endpoint by considering your suggestion. 7. Why were these data (K-M analysis) not presented through the survival curve? The survival curve serves exactly to see the recurrence time because in the table it is not clear. Table 3 is not adequate. Response: thank you, corrected accordingly. https://doi.org/10.1371/journal.pone.0277362.r004
26 Oct 2022 Decision Letter - José Luiz Fernandes Vieira, Editor Therapeutic efficacy of Chloroquine for the treatment of uncomplicated Plasmodium vivax infection in Shewa Robit, Northeast Ethiopia PONE-D-22-18161R2 Dear Dr. HABTAMU Belay Temesgen, 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, José Luiz Fernandes Vieira Academic Editor PLOS Reviewers' comments: All the suggestions were done by authors. https://doi.org/10.1371/journal.pone.0277362.r005
Formally Accepted
22 Nov 2022 Acceptance Letter - José Luiz Fernandes Vieira, Editor PONE-D-22-18161R2 Therapeutic efficacy of Chloroquine for the treatment of uncomplicated Plasmodium vivax infection in Shewa Robit, Northeast Ethiopia Dear Dr. Temesgen: 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 Dr. José Luiz Fernandes Vieira Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0277362.r006

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