Reviewer ’s report  
 
Title: Detecti on of bacterial DNA by in situ hybridization in patients with decompensated liver 
cirrhosis  
 
Version: 0 Date: 18 May 2017  
 
Reviewer: Koji Uchino  
 
Reviewer's report:  
 
In this study, the authors explored the usefulness of in situ hybridization (ISH) for detecting 
bacterial DNA among patients with decompensated cirrhosis. There are several problems in this 
manuscript for publication as described below.  
 
Major Comments  
1) Rationale of this study is unclear. The authors should discriminate symptomatic bacteremia 
and asymptomatic transient bacteremia which is not clinically important. Among patients 
who were not diagnosed as SBP by PMN counts, only one patients was positive for ISH 
(Figure 2). Thus, PMN count is adequate test for diagnosis of SBP. The usefulness of ISH in 
clinical setting should be clarify.  
2) It could be clarify how to select the patients who were tested by ISH. Of 60 patients who 
were positive for ISH, thirty patients were afebrile (Table 3). Were asymptomatic patients 
included in this study? Asymptomatic patients should be excluded because  asymptomatic 
transient bacteremia is not clinically important.  
 
Minor Comments  
1) The author should clearly describe that this study was designed retrospectively.  
2) Diagnostic criteria of decompensated LC and SBP should be described.  
3) The author sh ould be described whether blood samples were taken before and after injection 
of antibiotics. The timing of blood test affects the sensitivity of blood culture.  
4) Presence of ascites should be included in the factors of baseline characteristics.  
5) Six patients who were positive for ISH and blood culture should be excluded in the analysis 
of Table 2b because these patients overlapped between the two groups.  
Are the methods appropri ate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Unable to assess  
 
Are the c onclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistica l review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary 
from an  organisation that may in any way gain  or lose financially from the 
publication of this  manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the fut ure? 
 
3. Do you hold or are you currently applying for any patents relating to the content of 
the manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of  the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is acc epted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommon s.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title: Detecti on of bacterial DNA by in situ hybridization in patients with decompensated liver 
cirrhosis  
 
Version: 0 Date: 18 May 2017  
 
Reviewer: Koji Uchino  
 
Reviewer's report:  
 
In this study, the authors explored the usefulness of in situ hybridization (ISH) for detecting 
bacterial DNA among patients with decompensated cirrhosis. There are several problems in this 
manuscript for publication as described below.  
 
Major Comments  
1) Rationale of this study is unclear. The authors should discriminate symptomatic bacteremia 
and asymptomatic transient bacteremia which is not clinically important. Among patients 
who were not diagnosed as SBP by PMN counts, only one patients was positive for ISH 
(Figure 2). Thus, PMN count is adequate test for diagnosis of SBP. The usefulness of ISH in 
clinical setting should be clarify.  
2) It could be clarify how to select the patients who were tested by ISH. Of 60 patients who 
were positive for ISH, thirty patients were afebrile (Table 3). Were asymptomatic patients 
included in this study? Asymptomatic patients should be excluded because  asymptomatic 
transient bacteremia is not clinically important.  
 
Minor Comments  
1) The author should clearly describe that this study was designed retrospectively.  
2) Diagnostic criteria of decompensated LC and SBP should be described.  
3) The author sh ould be described whether blood samples were taken before and after injection 
of antibiotics. The timing of blood test affects the sensitivity of blood culture.  
4) Presence of ascites should be included in the factors of baseline characteristics.  
5) Six patients who were positive for ISH and blood culture should be excluded in the analysis 
of Table 2b because these patients overlapped between the two groups.  
Are the methods appropri ate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Unable to assess  
 
Are the c onclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistica l review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary 
from an  organisation that may in any way gain  or lose financially from the 
publication of this  manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the fut ure? 
 
3. Do you hold or are you currently applying for any patents relating to the content of 
the manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of  the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is acc epted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommon s.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Detecti on of bacterial DNA by in situ hybridization in patients with decompensated liver 
cirrhosis  
 
Version: 0 Date: 18 May 2017  
 
Reviewer: Koji Uchino  
 
Reviewer's report:  
 
In this study, the authors explored the usefulness of in situ hybridization (ISH) for detecting 
bacterial DNA among patients with decompensated cirrhosis. There are several problems in this 
manuscript for publication as described below.  
 
Major Comments  
1) Rationale of this study is unclear. The authors should discriminate symptomatic bacteremia 
and asymptomatic transient bacteremia which is not clinically important. Among patients 
who were not diagnosed as SBP by PMN counts, only one patients was positive for ISH 
(Figure 2). Thus, PMN count is adequate test for diagnosis of SBP. The usefulness of ISH in 
clinical setting should be clarify.  
2) It could be clarify how to select the patients who were tested by ISH. Of 60 patients who 
were positive for ISH, thirty patients were afebrile (Table 3). Were asymptomatic patients 
included in this study? Asymptomatic patients should be excluded because  asymptomatic 
transient bacteremia is not clinically important.  
 
Minor Comments  
1) The author should clearly describe that this study was designed retrospectively.  
2) Diagnostic criteria of decompensated LC and SBP should be described.  
3) The author sh ould be described whether blood samples were taken before and after injection 
of antibiotics. The timing of blood test affects the sensitivity of blood culture.  
4) Presence of ascites should be included in the factors of baseline characteristics.  
5) Six patients who were positive for ISH and blood culture should be excluded in the analysis 
of Table 2b because these patients overlapped between the two groups.  
Are the methods appropri ate and well described?  
If not, please specify what is required in your comments to the authors.  
 
No 
 
Does the work include the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Unable to assess  
 
Are the c onclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistica l review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have you in the past five years received reimbursements, fees, funding, or salary 
from an  organisation that may in any way gain  or lose financially from the 
publication of this  manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the fut ure? 
 
3. Do you hold or are you currently applying for any patents relating to the content of 
the manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of  the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial competing interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests.  
 
I agree to the open peer review policy of the journal. I understand that my name will be included  
on my report to the authors and, if the manuscript is acc epted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommon s.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant