Era of Molecular Diagnostics Techniques before and after the COVID-19 Pandemic

Round 1

Reviewer 1 Report

Not much care/attention has been paid to the write-up of the paper. A review should be more than a compilation of the results reported in the literature. It should, in fact, be a critical assessment of the present knowledge with some clear conclusions. The authors have done a superficial exercise. Except table 1, none of the tables is supported by references. I don’t know, the images are self-made or taken from other publications. Authors have collected testing kit names and say it as molecular techniques. There should be a broad and clear-cut categorization and explanation of techniques in terms of merit and demerit. Although table 2 is discussing some techniques but they are not well categorized. I do not see any significant information or novelty in this manuscript. At this stage, publication would be premature. Therefore, I must reject it.  

Author Response

Dear Editor and Reviewers

Thank you so much for reviewing the manuscript. We believe that the questions you raised will improve our manuscript for a broad readership in the field.

We tried our best to answer your comments and incorporate them into the manuscript. All the recommended changes are highlighted in yellow in the manuscript.

Please see the one-by-one answers for comments below

 

Reviewer 1

Not much care/attention has been paid to the write-up of the paper. A review should be more than a compilation of the results reported in the literature. It should, in fact, be a critical assessment of the present knowledge with some clear conclusions. The authors have done a superficial exercise. Except table 1, none of the tables is supported by references. I don’t know, the images are self-made or taken from other publications. Authors have collected testing kit names and say it as molecular techniques. There should be a broad and clear-cut categorization and explanation of techniques in terms of merit and demerit. Although table 2 is discussing some techniques but they are not well categorized. I do not see any significant information or novelty in this manuscript. At this stage, publication would be premature. Therefore, I must reject it.  

 

Response: Dear reviewer, thank you for your time in reviewing the manuscript. However, we would like to convince you of some of the questions you raised.

 

Question 1: Except table 1, none of the tables is supported by references

Response: Table one has been made by a collection of various data from a number of sources, and the sources are mentioned in the last column, but the other tables are not made by a collection of data from variable sources. So, there was no need to make a column for reference, but the source from which the table was made has been mentioned in the paragraph with the reference highlighted. For instance, “Table 2.1. CoV-19 Advance Diagnostic Techniques in Development (Sheridan, 2020).” Another example is “Table 2.2. List of kits for SARS Cov-19 approved by FDA 2021” whose data is taken directly from website of FDA and the reference is mentioned in the above paragraph “In August, 2021, a list of FDA approved many categories of kits varying on the principle such as RT-PCR kit , antigen detection kit, and antibody detection kit for detection of COVID-19 virus as shown in table 2.2 (FDA, 2021).”

 

Question 2: I don’t know, the images are self-made or taken from other publications

 

Response: The images are self-made with data collected from publications, and they are free from plagiarism.

 

Question 3: Authors have collected testing kit names and say it as molecular techniques

Response: The only table which says,

“Table 2.1. CoV-19 Advance Diagnostic Techniques in Development (Sheridan, 2020)”

has an error in the title only. It was supposed to be “SARS-CoV-2 advanced diagnostics in development”. The word technique has been added by mistake, which creates confusion as this table is directly taken from the reference mentioned. One can cross-check. We have changed the error point in the title. All other tables have clearly mentioned the difference in kits.

 

Question 4: There should be a broad and clear-cut categorization and explanation of techniques in terms of merit and demerit.

 

Response: There were demerits in the techniques used in early COVID-19, which have been mentioned in lines 151-160 shortly. While talking about the demerits of the latest techniques, these techniques are still in process, so there is limited or no data available about the demerits of the latest technologies.

 

Question 5: I do not see any significant information or novelty in this manuscript

 

Response: There is no such review available so far, to the best of our knowledge, which compares the timeline of molecular technique before and after COVID-19 starting from the millennia of Hippocrates. To the FDA, 2021 statistics for new kits developed for COVID-19. Either there are reviews that only list the advancements in molecular techniques (names only) or explain the techniques for the detection of COVID-19 alone. But this review is written like a story starting from the Hippocrates era to techniques early used in COVID-19 and why their limitations led to the development of novel techniques for COVID-19.

 

Author Response File: Author Response.docx

Reviewer 2 Report

Authors have presented a good explanation of changes in molecular diagnostic techniques before and after covid-19 pandemic. They have  detailed a brief explanation of techniques from the past to the present. They have compiled the origin, history, and simple working mechanism for the molecular diagnostic tests. Rapid growth of molecular diagnostic techniques during and after covid-19 period have been emphasized.

 Some suggestions for authors:

 

 1) The paper contains a lot of abbreviations. It would be wise to provide a comprehensive list of abbreviations.

 

 2)  Line 201, …1fg/ml and 100fg/ml … might be a typo. It may be a range, 1fg/ml - 100fg/ml.

 

 3) Line 217, …confined in nanoparticles. Please specify what material nanoparticles are these. Gold nanoparticles?

 

 4) Line 225, LOD of 0.2pM. Space between the value and unit is missing. This has been seen in many other places as well.

 

 5)  A brief explanation of the working of MIP sensor (Line 317) would make this section clearer. As described for other techniques, add details on this as well.

 

 6)  All the rapid test kits are supposed to give the results fast. For the tables 3, 4 and 5, if authors could add a column with time the kit gives result, it would be a good comparison.

 

Author Response

Dear Editor and Reviewers

Thank you so much for reviewing the manuscript. We believe that the questions you raised will improve our manuscript for a broad readership in the field.

We tried our best to answer your comments and incorporate them into the manuscript. All the recommended changes are highlighted in yellow in the manuscript.

Please see the one-by-one answers for comments below.

 

Reviewer 2

Authors have presented a good explanation of changes in molecular diagnostic techniques before and after covid-19 pandemic. They have  detailed a brief explanation of techniques from the past to the present. They have compiled the origin, history, and simple working mechanism for the molecular diagnostic tests. Rapid growth of molecular diagnostic techniques during and after covid-19 period have been emphasized.

 Some suggestions for authors:

 

1) The paper contains a lot of abbreviations. It would be wise to provide a comprehensive list of abbreviations.

Response: A list of abbreviations has been added to the main manuscript.

 

 2)  Line 201, …1fg/ml and 100fg/ml … might be a typo. It may be a range, 1fg/ml - 100fg/ml.

Response: The Typo mistake corrected

 

 3) Line 217, …confined in nanoparticles. Please specify what material nanoparticles are these. Gold nanoparticles?

 Response: Yes, it is gold nanoparticles, edited.

 

 4) Line 225, LOD of 0.2pM. Space between the value and unit is missing. This has been seen in many other places as well.

Response: This kind of typo mistake made clear in the whole manuscript

 

 5)  A brief explanation of the working of MIP sensor (Line 317) would make this section clearer. As described for other techniques, add details on this as well.

Response:  The brief explanation of the MIP sensor has been added.

 

 6)  All the rapid test kits are supposed to give the results fast. For the tables 3, 4 and 5, if authors could add a column with time the kit gives result, it would be a good comparison

Response: For sure, it would have been a very good comparison. The data of these tables have been taken directly from FDA statistics for 2021, which doesn’t mention the time of these kits. Hence, we are unable to make the column of time as there is no data available on the internet.

 

Author Response File: Author Response.docx

Reviewer 3 Report

This is a review of COVID-19 molecular assays that have been developed. There are several unique technologies I was unaware of and should be interesting to readers.

Major comments:

1. The background on PCR seems a bit extensive and unnecessary, but is fine to include if the authors feel it necessary. If the authors are going to present the thorough recitation of previous advances, I would recommend they cite the actual reports and not just use "catch all" citations like Demidov 2003.

2. Some of the described technologies of antigen and antibody diagnostics aren't strictly molecular from the traditional sense, but as protein targets they are suitable for inclusion. In this respect, I would also recommend adding the studies that include a Nature Biotechnology paper that uses Mass Spec to detect SARS-CoV-2 as well as some other biosensors such as breath detection technology. 

3. Please be sure citations are correctly attributed to relevant studies. For example on line 196, the only citation for Biosensors is a 2018 article when they specifically say that another technology was applied to SARS-CoV-2 (not present at the time of 2018).

4. Line 226- please define 'high precision' this is a bit vague here.

 

Author Response

Dear Editor and Reviewers

Thank you so much for reviewing the manuscript. We believe that the questions you raised will improve our manuscript for a broad readership in the field.

We tried our best to answer your comments and incorporate them into the manuscript. All the recommended changes are highlighted in yellow in the manuscript.

Please see the one-by-one answers for comments below.

 

Reviewer 3

This is a review of COVID-19 molecular assays that have been developed. There are several unique technologies I was unaware of and should be interesting to readers.

Major comments:

1. The background on PCR seems a bit extensive and unnecessary, but is fine to include if the authors feel it necessary. If the authors are going to present the thorough recitation of previous advances, I would recommend they cite the actual reports and not just use "catch all" citations like Demidov 2003.

Response: We have only focused on actual reports for citations in this review; unrelated citations are removed.

2. Some of the described technologies of antigen and antibody diagnostics aren't strictly molecular from the traditional sense, but as protein targets they are suitable for inclusion. In this respect, I would also recommend adding the studies that include a Nature Biotechnology paper that uses Mass Spec to detect SARS-CoV-2 as well as some other biosensors such as breath detection technology. 

Response: Dear reviewer, this review has focused on summaries of the major molecular biology techniques before and after COVID-19. As this review focused on major biosensors to give an idea to the readers about the type of advancements in molecular techniques in this era. Many other minor biosensors are also present e,g. Piezoelectric immunosensor, thermal biosensors, Electrochemiluminescence-Based Biosensors, Photoluminescence-Based Biosensors, Nanoplasmonic-Based Biosensors, Quartz Crystal Microbalance Based Biosensors, Reactive Oxygen Species based biosensors, etc. If we add all the possible details techniques and their details, it will create huge data, which will be a little confusing/or frustrating to understand. We are planning to write an article covering all the types of biosensors for COVID-19 only.

3. Please be sure citations are correctly attributed to relevant studies. For example on line 196, the only citation for Biosensors is a 2018 article when they specifically say that another technology was applied to SARS-CoV-2 (not present at the time of 2018).

Response: There might be a misunderstanding in the way the biosensors have been explained, which is giving an impact that Biosensors were only innovated for Covid-19 in 2020. I am going to elaborate further and explain clearly in that paragraph so miscomprehension doesn’t occur. Biosensors have been innovated before corona for detecting other diseases, but after COVID-19, this technology was optimized for the detection of viral coronavirus, making that optimization as a novel. We have modified the lines in the given paragraph so the reader doesn’t get confused.

4. Line 226- please define 'high precision' this is a bit vague here.

Response: The higher precision here describes that the LSPR sensors are capable of differentiating the SARS-CoV and SARS-CoV2 with 96% precision accuracy.

Author Response File: Author Response.doc

Round 2

Reviewer 1 Report

I rejected this manuscript in the previous round of review. I do not see any significant information or novelty in this manuscript, therefore, my decision is same and I don't want to review it further.