PONE-D-19-32764

Towards low cost, multiplex clinical genotyping: 4 fluorescent Kompetitive Allele-Specific PCR and its application on pharmacogenetics

PLOS ONE

Dear Mrs. Lin,

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We look forward to receiving your revised manuscript.

Kind regards,

Ruslan Kalendar, PhD

Academic Editor

PLOS ONE

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https://doi.org/10.1373/clinchem.2008.115295

https://doi.org/10.1016/j.jmoldx.2017.05.007

In your revision ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the Methods section. Further consideration is dependent on these concerns being addressed.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Yes

Reviewer #2: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: No

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3. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: No

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #2: No

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5. Review Comments to the Author

Reviewer #1:

This paper describes an incremental advance that is unlikely to be adopted because there are many better and more accessible methods for genotyping. While the work demonstrates multiplexed genotyping, the level of multiplexing is very limited.

Specific comments:

Too few samples with rare genotypes are tested. It is important to show that there are no errors with rare genotypes.

The statement in the materials and methods "Sequence of used primers is available upon request" is not acceptable for PLOS One. Authors should include all sequence information in the paper or Supplementary Information section. It is not optional information that should have to be requested.

English language needs to be reviewed by a native English speaker.

Reviewer #2:

In the current study, a modified Kompetitive Allele Specific PCR, in which 4 fluorescents were used, was described. Further, this modified KASP was used and evaluated. Overall, this job is an improvement for the routine KASP that can be a cost-effective high-throughput SNP genotyping alternative for pharmacogene variants. There are many mistakes in the text concerning English wording and grammar. It is suggested to authors to co-opt a native English speaker when revising the manuscript.

Specific Comments

1. Why did rs1057910 and rs4986893 were selected to evaluated the modified KASP?

2. the result of modified KASP for clinical samples was in 100% agreement with that from hospital report. The authors should describe what kind of method the hospital reports based on. Moreover, the number of the samples (n=40) that were carried out to evaluate the mothed was extremely limited. The authors should increase sample size.

3. Three-line table should be used in the manuscript.

4. The format of reference in the text doesn’t meet the standard of PLOS ONE.

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Reviewer #1: No

Reviewer #2: No

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Towards low cost, multiplex clinical genotyping: 4-fluorescent Kompetitive Allele-Specific PCR and its application on pharmacogenetics

PONE-D-19-32764R1

Dear Dr. Lin,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

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With kind regards,

Ruslan Kalendar, PhD

Academic Editor

PLOS ONE