An In Silico Analysis of PCR-Based Monkeypox Virus Detection Assays: A Case Study for Ongoing Clinical Surveillance
, Hayden N. Brochu 1, Qimin Zhang 1, Jonathan D. Williams 2 and Lakshmanan K. Iyer 1,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis is a topical and relevant study with immediate implications for MPXV detection but also wider applications for primer evaluation for development of other PCR-based clinical tests.
The paper presents a data-rich topic clearly, supported by informative graphics. I have no specific comments to add or recommended amendments.
Author Response
Dear reviewer,
Thank you so much for your time and effort in reviewing our manuscript, we greatly appreciated it!
Reviewer 2 Report
Comments and Suggestions for AuthorsThe authors examined 5,210 Monkeypox virus (MPXV) genomes sourced from the NCBI Viral Genomes resource (accessed on 05/21/2023) and obtained lineage information from both the Nextstrain hMPXV (NC_063383.1) and MPXV (NC_003310.1) databases. The primary focus of the study was the evaluation of 18 PCR primer sets explicitly designed to target MPXV.
The results of the study revealed that only five primer sets yielded nearly perfect matches against the targeted MPXV lineages. Conversely, the remaining primer sets all exhibited 1-2 mismatches against almost all MPXV lineages.
I greatly appreciate your study, which is both interesting and well-described. Just one question: how much can the choice of sample type (skin lesions, throat, blood, stools) and the timing of collection (e.g., early infection versus recovery phase) impact the sensitivity and specificity of these primers?
Author Response
Please see the attachment. Thank you.
Author Response File: 
Author Response.docx
