Merkel cell polyomavirus is implicated in a subset of Merkel cell carcinomas, in the Indian subcontinent

Arora, Reety; Rekhi, Bharat; Chandrani, Pratik; Krishna, Sudhir; Dutt, Amit

doi:10.1016/j.micpath.2019.103778

Cited by 6 publications

(2 citation statements)

References 51 publications

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“…For examples, CM2B4 often gives non-specific staining in both tonsillar and lymphoid tissues (Leroux-Kozal et al, 2015;Moshiri et al, 2017). Other antibodies that detect MCV T antigens, such as Ab3 and Ab5 have been reported, however none have been used as frequently as CM2B4 for detecting viral proteins in the clinic (Cheng et al, 2013;Harms et al, 2018;Arora et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

“…Both viral positive and viral negative Merkel tumors show identical phenotypes, immunohistochemical (IHC) staining patterns and occur at similar sun-exposed locations on the human body (Hodgson, 2005;Arora et al, 2012Arora et al, , 2019DeCaprio, 2017;Moore and Chang, 2017;Moshiri et al, 2017;Harms et al, 2018). A pathologist is unable to distinguish between the two types of tumors without the aid of Next-Gen-Sequencing (Goh et al, 2016;Harms et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

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DETECTing Merkel Cell Polyomavirus in Merkel Tumors

Arora

Gupta

Vijaykumar

et al. 2020

Front. Mol. Biosci.

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Merkel cell carcinoma (MCC) is a rare, aggressive skin cancer caused either by Merkel cell polyomavirus (MCV) T antigen expression, post-integration (∼80% cases), or by UV-mediated DNA damage. Interestingly, overall survival of MCV-positive Merkel cell carcinoma patients is better, making this differential information of significant diagnostic and prognostic value. Also, MCV provides a direct target for therapy in MCC patients. Currently, the methods used for diagnosis of MCV in tumors are often discordant and unreliable. Here we used a guided molecular scissors based-DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) technique to develop an in vitro molecular diagnostic tool for MCV-positive MCC. DETECTR couples recombinase polymerase based amplification of target MCV DNA with Cas12a mediated detection. CRISPR diagnostics couple specific detection followed by cutting of the pathogenic DNA by the Cas enzyme-gRNA complex, with non-specific cutting of ssDNA that provides a measurable visual cue. To detect MCV DNA in MCC, we designed Cas12a gRNAs targeting the MCV DNA and tested their targeting efficiency, and sensitivity using a fluorophore quencher labeled reporter assay. We show that MCV DETECTR system can detect MCV integrated in Merkel tumor rapidly, specifically and with femto-molar sensitivity. Our study is a preliminary, proof-of-principle analysis showing the use of CRISPR for MCV diagnosis. Further validation in human tumor samples is needed for its clinical use in the near future. This new system is promising and we hope it can be coupled with immunohistochemical studies to diagnose the viral status of MCC in clinics soon.

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