Progression of LAMP as a Result of the COVID-19 Pandemic: Is PCR Finally Rivaled?
Round 1
Reviewer 1 Report
1. I would suggest including better resolution images for figures 3-7, if possible.
2. As shown in Table 1, the sensitivity of the LAMP reaction is described, and related descriptions are also given below the table, in which a comparison with PCR is mentioned. If possible, it is suggested to add a relevant comparison between the existing LAMP and PCR methods of the Convid-19 detection, which can be presented in table form.
3. In section 3.4, the author have a brief review of the all-in-one LAMP system, However, the all-in-one system based on microfluidic technology (such as the digital microfluidic system) in the current study has also been widely applied in the detection of COVID-19 based on LAMP method, and has shown obvious advantages. It is suggested that the author add relevant brief discussion to the review.
4. In figure 3, such as figure C also include the original figure number B), and figure D also can see the original name fig 1 and fig 2. The original name will make a misunderstanding of the figure, if possible, please revised it.
Author Response
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
The reviewed manuscript is decicated to LAMP in the aspect of COVID-19 pandemic. Authors described the state of affairs in the application and development of new LAMP-based tests for SARS-CoV-2 testing. The review is timely and provides an interesting survey for researchers in the field of molecular diagnostics. However, a few comments need to be made concerning several issues:
1. Page 1 lanes 35-36 “Since most viruses are RNA viruses, including SARS-CoV-2, RNA must be converted to complementary DNA” – it would be better to reformulate that statement as RNA viruses are not the most prevalent for all domains of life.
2. Page 1 lanes 43 “while PCR typically requires three temperatures” – commonly using TaqMan-based PCR tests are based on cycling of 2 temperatures.
3. Page 2, lane 73 “LAMP is far more prone to “false positives.”” – while LAMP is prone to false positive results, it also have several other limitations, e.g. the need for conservative sites for 4-6 primers whisch is tricky for highly diverse viruses.
4. Page 4, lane 127 “In terms of point-of-care diagnostics, LAMP holds the most potential for portability” – other methods of isothermal amplification also could be use for lab-on-chip devices.
5. Page 6, lane 202 “Numerous fluorescent microplate readers and even real-time LightCyclers originally” – it would be better to change “LightCyclers” to “PCR machines”.
6. Tables with specs of various LAMP-based devices would improve the readability of the manuscript.
7. A paragraph describing various approaches for the detection of LAMP results would be highly appreciated.
Author Response
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Author Response File: Author Response.pdf
Reviewer 3 Report
See the file
Comments for author File: Comments.pdf
Author Response
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Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report
The authors improved the article a lot. However, I would still suggest some minor to major revisions on the resubmitted article.
Introduction:
* "and cost of the RT-PCR test -" (add the -)
* 'at the peak... some areas' --> RT-PCR has a relative long assay time as is stated in the previous sentence, but the time-toresult being up to two weeks is caused by other things. Elaborate on this.
* 'two to three' and 'four to six' (remove the -'s)
* combined with simple visual detection methods (instead of paired)
* 'proven to increase LAMP specificity and sensitivity' --> clinical or analytical sensitivity in this case?
Section 2
* Fraction of cases detected is not (exactly) the same as the clinical sensitivity. Please clarify/adjust.
Section 3
* Split figure 3 in multiple figures. The subfigures are too small to be read properly.
* When multiple sentences after each other have the same reference, the reference can be put there once (at the end of the last sentence).
* SYBR Green is only inhibitory when the concentration is too high.
* What is the drawback of colorimetric detection? What about the sensitivity compared to other methods?
Later on in the article the sensitivy is discussed, which is good. But I miss an "introduction" sentence which explains what is going to be discussed in this section.
* Also figure 4 is quite crowded. Only keep the parts of the subfigures that are of interest. Also refer to the subfigures in the text.
* Figure 5: Again a very crowded picture; the steps are not readable. Please adapt the figure.
* There are multiple sources that state that the ID NOW makes use of the NEAR technique.
- https://pubs.rsc.org/en/content/articlelanding/2021/AY/D1AY00947H
- https://www.cell.com/iscience/fulltext/S2589-0042(20)30596-4?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS2589004220305964%3Fshowall%3Dtrue
- https://www.sciencedirect.com/science/article/pii/S259013702200053X
* Table 2 is a nice addition
* Figure 6 is also too crowded. Please adapt.
Section 4:
* 'RT-PCR yelds a decent overall clinical sensitivity' --> for which test? or mention according to whom. The statement as it is now is too bold. Also rephrase 'LAMP... yielded a clinical sensitivity of 65%'
Section 5:
* Some headers are still quite long. Only conclusion would be fine by me.
* Why are antigen tests unreliable?
* Do not introduce new information in the conclusion (e.g. the work of Pu et al.)
* What is meant with 'false rate'. More information is needed to place the 6-12% rate. In comparison to what for instance? Which tests?
Author Response
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Author Response File: Author Response.pdf
Round 3
Reviewer 3 Report
* check the sentence: has also been proven to increase LAMP
analytical specificity and sensitivity and specificity [33] and clinical specificity and sensitivity [34-35].
* pictures are still quite crowded
Author Response
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Author Response File: Author Response.pdf
