Biomarkers in the Diagnosis of Aspergillosis

Round 1

Reviewer 1 Report

This review article provides a detailed overview of established and emerging biomarkers for the diagnosis of aspergillosis. The authors covered major clinical subtypes of aspergillosis and their diagnostic tools. Please find my comments below.

Major comments

1. The authors should discuss why the detection of biomarkers differs from one patient population to another. They can also add a summary paragraph at the end of each biomarker paragraph, which includes the strengths and weaknesses of biomarkers and how to prioritize them in clinical settings.
2. The paragraphs on Triacetylfusarinine C and volatile organic compounds are very informative. However, the authors did not address the challenges of biomarkers, such as cost, technical difficulty, lack of standardization, and regulatory issues. The authors can describe these biomarkers as experimental or emerging and discuss what is needed for them to be used clinically.
3. Although antifungal resistance is mentioned as a health challenge, the authors did not fully explore the role of biomarkers in resistance detection, especially azole resistance using CYP51A mutations. They can expand the PCR paragraph to include resistance detection, its clinical implications, and current challenges.

 

 

The paragraph (lines 253-261) should be revised to remove repeated sentences.

Author Response

Reviewer #1

This review article provides a detailed overview of established and emerging biomarkers for the diagnosis of aspergillosis. The authors covered major clinical subtypes of aspergillosis and their diagnostic tools. Please find my comments below.

Major comments

1. The authors should discuss why the detection of biomarkers differs from one patient population to another. They can also add a summary paragraph at the end of each biomarker paragraph, which includes the strengths and weaknesses of biomarkers and how to prioritize them in clinical settings.
2. Thank you for your suggestion; it is very important. We have created the figure based on your recommendations.
3. The paragraphs on Triacetylfusarinine C and volatile organic compounds are very informative. However, the authors did not address the challenges of biomarkers, such as cost, technical difficulty, lack of standardization, and regulatory issues. The authors can describe these biomarkers as experimental or emerging and discuss what is needed for them to be used clinically.
4. We have added a statement following your suggestion.
5. Although antifungal resistance is mentioned as a health challenge, the authors did not fully explore the role of biomarkers in resistance detection, especially azole resistance using CYP51A They can expand the PCR paragraph to include resistance detection, its clinical implications, and current challenges.
6. We have added a statement following your suggestion.

 

Detailed Comments:

The paragraph (lines 253-261) should be revised to remove repeated sentences.

1. Thank, removed the duplicate sentence.

Reviewer 2 Report

The manuscript entitled “Biomarkers in the Diagnosis of Aspergillosis” addresses an important and clinically relevant topic. However, the manuscript contains numerous inaccuracies in the extraction and interpretation of information from the cited references. These issues significantly undermine the scientific reliability of the review and limit its value to readers. Overall, a meaningful and balanced comparison between diagnostic methods is lacking, and several biomarkers are discussed with inaccurate or misleading conclusions.

Detailed Comments:

1. Lines 173–186:
   This section consists of a very long paragraph that relies entirely on a single reference and is not sufficiently specific to the topic. Several key studies and review articles are missing. This section should be substantially revised to clearly present and compare the diagnostic importance, sensitivity, and specificity of each antibody-based assay.
2. Repeated paragraph (Lines 253–257 and 257–261):
   The same paragraph is duplicated while citing two different references (references 10 and 39). This must be corrected. The repeated text is:

“GM detection has also been evaluated in patients with CPA. Serum ELISA has shown low sensitivity and specificity, even when the index cutoff value is reduced from 1.5 to 0.5 (the recommended cutoff for the Platelia Aspergillus Ag assay, Bio-Rad). Studies have reported a BAL sensitivity and specificity of 77%, slightly higher than those observed in serum (10).”

1. Lines 280–283:
   The manuscript refers to reference (29) as a meta-analysis on mannoproteins; however, this reference is neither a meta-analysis nor does it discuss mannoproteins. This represents a major error.
2. Lines 331–341:
   This long paragraph discusses the limitations of the BDG biomarker while citing only a single reference that does not contain the stated information. This is a significant mistake and requires correction with appropriate supporting references.
3. Lines 407–409:
   The authors say:

“In an initial study, a sensitivity of 81% and a specificity of 90% were reported in urine samples from patients with hematological malignancies.”
However, the cited study was conducted in mice, not in patients, and does not support these results.

1. Lines 437–439:
   The authors mention sensitivity and specificity values of 80–100% for a diagnostic technique but cite only a review article, rather than original studies. Primary research articles should be cited to support such claims.
2. Lines 443–446:
   The claim regarding increased sensitivity from 60% to 83% in hematological patients through combined GM and BDG detection is not accurately supported by the cited references and should be revised and corrected.
3. The manuscript does not mention the PNA-FISH method for Aspergillus identification, which is a relevant diagnostic approach and should be discussed.
4. The page numbers are incorrect and should be carefully revised.

Author Response

Reviewer #2

The manuscript entitled “Biomarkers in the Diagnosis of Aspergillosis” addresses an important and clinically relevant topic. However, the manuscript contains numerous inaccuracies in the extraction and interpretation of information from the cited references. These issues significantly undermine the scientific reliability of the review and limit its value to readers. Overall, a meaningful and balanced comparison between diagnostic methods is lacking, and several biomarkers are discussed with inaccurate or misleading conclusions.

R. Dear Reviewer. We sincerely apologize. The reference management software we used became corrupted, which caused most of citations to shift by one number. As a result, the cited references did not correspond correctly with the text. This issue has now been corrected, and all comments related to the references have been fully addressed.

Detailed Comments:

Lines 173–186:
This section consists of a very long paragraph that relies entirely on a single reference and is not sufficiently specific to the topic. Several key studies and review articles are missing. This section should be substantially revised to clearly present and compare the diagnostic importance, sensitivity, and specificity of each antibody-based assay.

R. Reference corrected.

Repeated paragraph (Lines 253–257 and 257–261):
The same paragraph is duplicated while citing two different references (references 10 and 39). This must be corrected. The repeated text is: “GM detection has also been evaluated in patients with CPA. Serum ELISA has shown low sensitivity and specificity, even when the index cutoff value is reduced from 1.5 to 0.5 (the recommended cutoff for the Platelia Aspergillus Ag assay, Bio-Rad). Studies have reported a BAL sensitivity and specificity of 77%, slightly higher than those observed in serum (10).”

R. Reference corrected.

Lines 280–283:
The manuscript refers to reference (29) as a meta-analysis on mannoproteins; however, this reference is neither a meta-analysis nor does it discuss mannoproteins. This represents a major error.

R. Reference corrected.

Lines 331–341:
This long paragraph discusses the limitations of the BDG biomarker while citing only a single reference that does not contain the stated information. This is a significant mistake and requires correction with appropriate supporting references.

R. Reference corrected.

Lines 407–409:
The authors say: “In an initial study, a sensitivity of 81% and a specificity of 90% were reported in urine samples from patients with hematological malignancies.”
However, the cited study was conducted in mice, not in patients, and does not support these results.

R. Reference corrected.

Lines 437–439:
The authors mention sensitivity and specificity values of 80–100% for a diagnostic technique but cite only a review article, rather than original studies. Primary research articles should be cited to support such claims.

R. Reference corrected.

Lines 443–446:
The claim regarding increased sensitivity from 60% to 83% in hematological patients through combined GM and BDG detection is not accurately supported by the cited references and should be revised and corrected.

1. Reference corrected.

The manuscript does not mention the PNA-FISH method for Aspergillus identification, which is a relevant diagnostic approach and should be discussed.

R. Thank you for your comment. We have now added a paragraph describing the PNA-FISH method

The page numbers are incorrect and should be carefully revised.

Revised.

Reviewer 3 Report

The review is generally comprehensive regarding the biomarkers used in Aspergillus infections. However, the authors could include a discussion of more useful protocols that clinicians could request in cases of invasive aspergillosis. The authors did not include a discussion section, which would improve the review by recommending specific biomarkers for each type of Aspergillus infection.

 

The authors could add a figure or image showing the types of Aspergillus infections, and it could also indicate which biomarkers might be most useful for each case.

Author Response

Reviewer #3

The review is generally comprehensive regarding the biomarkers used in Aspergillus infections. However, the authors could include a discussion of more useful protocols that clinicians could request in cases of invasive aspergillosis. The authors did not include a discussion section, which would improve the review by recommending specific biomarkers for each type of Aspergillus infection.

Detailed Comments:

The authors could add a figure or image showing the types of Aspergillus infections, and it could also indicate which biomarkers might be most useful for each case.

R. Thank you for the suggestion. We have added a figure summarizing the most relevant biomarkers and their clinical application.

Round 2

Reviewer 2 Report

The updated version of the manuscript continues to misextract information from the cited papers, which significantly affects the conclusion. 

The updated version of the manuscript continues to misextract information from the cited papers, which significantly affects the conclusion. 

Author Response

We conducted a detailed review. We identified that the Mendeley citation manager was disorganizing the references; therefore, we disabled it and inserted all citations manually.

Round 3

Reviewer 2 Report

I think the authors addressed the raised points 

I think the authors addressed the raised points 

Author Response

Thank you for your review！