Improved detection suggests all Merkel cell carcinomas harbor Merkel polyomavirus

Rodig, Scott J.; Cheng, Jingwei; Wardzala, Jacek; DoRosario, Andrew; Scanlon, Jessica J.; Laga, Alvaro C.; Martinez-Fernandez, A.; Barletta, Justine A.; Bellizzi, Andrew M.; Sadasivam, Subhashini; Holloway, Dustin T.; Cooper, Dylan J.; Kupper, Thomas S.; Wang, Linda C.; DeCaprio, James A.

doi:10.1172/jci64116

Cited by 194 publications

(193 citation statements)

References 48 publications

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“…DNA quantification was performed by Qubit 3.0 Fluorometer (Invitrogen, Carlsbad, CA) and diluted if necessary. 15 ng input DNA was analyzed by qPCR on the StepOne real-time PCR system (Applied Biosystems, Foster City, CA) using Taqman detection with the previously reported LT2 (LTAg) primer/probe set and SET9 (sTAg) primer/probe set (14). RNaseP TaqMan copy number reference assay (Thermo Fisher Scientific, Waltham, MA) was used to normalize results and confirm adequacy of DNA for PCR.…”

Section: Dna Extraction and Quantitative Pcrmentioning

confidence: 99%

“…qPCR is considered the most sensitive assay in common use for detection of MCPyV, but it has several limitations. Evaluation of multiple amplicons is necessary for maximal sensitivity, as neither LTAg nor sTAg detection alone is fully sensitive (14). In addition, low MCPyV qPCR signal may be detected due to infection by wild-type virus in normal tissues (15), and qPCR does not allow for direct correlation with morphology to confirm that the signal is originating in tumor cells.…”

Section: Introductionmentioning

confidence: 99%

“…The most commonly used antibody is CM2B4, which specifically detects MCPyV LTAg and not other polyomavirus T-antigens (9)(10)(11)(12)(13). Multiple reports have found IHC with CM2B4 antibody to be less sensitive than qPCR (14,16,17). Existing assays have limitations for routine clinical practice, warranting the need for development of new MCPyV detection methods.…”

Section: Introductionmentioning

confidence: 99%

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Age and Gender Associations of Virus Positivity in Merkel Cell Carcinoma Characterized Using a Novel RNA In Situ Hybridization Assay

Wang

Harms

Palanisamy

et al. 2017

Clinical Cancer Research

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Purpose-Merkel cell carcinoma (MCC) is a highly aggressive neuroendocrine tumor of the skin. Merkel cell polyomavirus (MCPyV) plays an oncogenic role in the majority of MCCs. Detection of MCPyV in MCCs has diagnostic utility and prognostic potential. We investigated whether RNAscope, an RNA in situ hybridization (ISH) assay for detection of RNA transcripts in tissues, is useful for MCPyV detection.Experimental Design-We applied an RNAscope probe targeting MCPyV T antigen transcripts on tissue microarrays (TMAs) and whole tissue sections encompassing 87 MCCs from 75 patients, 14 carcinomas of other types, and benign tissues. For comparison, quantitative PCR (qPCR) was performed on 57 cases of MCC from 52 patients.Results-RNA-ISH demonstrated the presence of MCPyV in 37/75 (49.3%) cases. Notably, tumors from younger patients (< 73 years) had a significantly higher virus positivity than those from elderly patients (≥ 73 years) (64.9% vs. 34.2%, P =0.011). Female patients had a higher positive rate of MCPyV than male patients (66.7% vs. 39.6%, P =0.032). Data from both RNA- HHMI Author Manuscript HHMI Author Manuscript HHMI Author Manuscriptdetermining MCPyV status, RNAscope had 100% sensitivity and 100% specificity. There was a strong correlation between qPCR copy number and RNA-ISH product score (Spearman's correlation coefficient R 2 = 0.932, P < 0.0001).Conclusions-RNA-ISH is comparably sensitive to qPCR for detection of MCPyV and allows for correlation with tissue morphology. This study also reveals a significant association between age, gender, and MCPyV positivity.

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Section: Dna Extraction and Quantitative Pcrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Age and Gender Associations of Virus Positivity in Merkel Cell Carcinoma Characterized Using a Novel RNA In Situ Hybridization Assay

Wang

Harms

Palanisamy

et al. 2017

Clinical Cancer Research

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“…In 2012, Rodig et al 17 was able to detect the presence of Merkel cell polyomavirus in nearly all cutaneous Merkel cell carcinomas using a novel monoclonal antibody (Ab3) and quantitative PCR. 49 The increased sensitivity of detection is likely related to shorter amplicons of PCR reactions, more primer sets, and utilization of quantitative PCR, which is more sensitive than the agarose-gel-based detection method.…”

Section: Possible Histogenesis Of Nodal Merkel Cell Carcinomamentioning

confidence: 99%

Merkel cell carcinoma of lymph node with unknown primary has a significantly lower association with Merkel cell polyomavirus than its cutaneous counterpart

Pan

Chen

et al. 2014

Modern Pathology

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Rare cases of Merkel cell carcinoma have been encountered in lymph nodes with unknown extranodal primary, which exhibit similar morphologic and immunophenotypic features to those in primary cutaneous Merkel cell carcinomas. However, it is uncertain whether the nodal Merkel cell carcinoma is a primary tumor of the lymph node or represents a metastasis from an occult or regressed extranodal lesion. To establish an accurate diagnosis of the nodal Merkel cell carcinoma can be challenging because of significant morphologic mimics, including lymphoblastic lymphoma and metastatic small cell carcinoma. Moreover, there is no consensus for a diagnostic term, and many different terms have been used, which can be confusing and may not fully reflect the nature of nodal Merkel cell carcinoma. In this study, we investigated the detailed clinicopathologic features of 22 nodal Merkel cell carcinomas, with comparison to 763 primary cutaneous cases retrieved from the literature. Overall, the nodal and cutaneous Merkel cell carcinomas shared similar clinical presentations, morphologic spectrum, and immunophenotype; both were mostly seen in elderly male with a typical neuroendocrine morphology. Most of cases expressed CK20, synaptophysin, and chromogranin A; and PAX5 and TdT were also positive in majority of cases. However, nodal Merkel cell carcinomas had a significantly lower association with Merkel cell polyomavirus than cutaneous cases (31% vs 76%, P ¼ 0.001). Therefore, these two entities may arise from overlapping but not identical biological pathways. We also recommend the use of the diagnostic term 'Merkel cell carcinoma of lymph node' to replace many other names used.

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“…5,18 Although numerous assays for Merkel cell polyomavirus detection have been reported, PCR targeting small T antigen and/or the 5' region of large T antigen is commonly used and highly sensitive. [19][20][21] Although present in the majority of Merkel cell carcinoma, Merkel cell polyomavirus has not been commonly observed in other malignancies. In particular, the presence of Merkel cell polyomavirus is highly specific for Merkel cell carcinoma relative to small cell lung carcinoma, 12,[22][23][24] and hence might be considered as a diagnostic marker in challenging cases including cytokeratin 20-negative Merkel cell carcinoma.…”

mentioning

confidence: 99%

Cytokeratin 20-negative Merkel cell carcinoma is infrequently associated with the Merkel cell polyomavirus

et al. 2015

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Merkel cell carcinoma is a rare, highly aggressive cutaneous neuroendocrine carcinoma most commonly seen in sun-damaged skin. Histologically, the tumor consists of primitive round cells with fine chromatin and numerous mitoses. Immunohistochemical stains demonstrate expression of neuroendocrine markers. In addition, cytokeratin 20 (CK20) is expressed in B95% of cases. In 2008, Merkel cell carcinoma was shown to be associated with a virus now known as Merkel cell polyomavirus in B80% of cases. Prognostic and mechanistic differences between Merkel cell polyomavirus-positive and Merkel cell polyomavirus-negative Merkel cell carcinoma may exist. There has been the suggestion that CK20-negative Merkel cell carcinomas less frequently harbor Merkel cell polyomavirus, but a systematic investigation for Merkel cell polyomavirus incidence in CK20-negative Merkel cell carcinoma has not been done. To test the hypothesis that Merkel cell polyomavirus is less frequently associated with CK20-negative Merkel cell carcinoma, we investigated 13 CK20-negative Merkel cell carcinomas from the files of the Cleveland Clinic and the University of Michigan for the virus. The presence or absence of Merkel cell polyomavirus was determined by quantitative PCR performed for Large T and small T antigens, with sequencing of PCR products to confirm the presence of Merkel cell polyomavirus. Ten of these (77%) were negative for Merkel cell polyomavirus and three (23%) were positive for Merkel cell polyomavirus. Merkel cell polyomavirus is less common in CK20-negative Merkel cell carcinoma. Larger series and clinical follow-up may help to determine whether CK20-negative Merkel cell carcinoma is mechanistically and prognostically unique.

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Improved detection suggests all Merkel cell carcinomas harbor Merkel polyomavirus

Cited by 194 publications

References 48 publications

Age and Gender Associations of Virus Positivity in Merkel Cell Carcinoma Characterized Using a Novel RNA In Situ Hybridization Assay

Age and Gender Associations of Virus Positivity in Merkel Cell Carcinoma Characterized Using a Novel RNA In Situ Hybridization Assay

Merkel cell carcinoma of lymph node with unknown primary has a significantly lower association with Merkel cell polyomavirus than its cutaneous counterpart

Cytokeratin 20-negative Merkel cell carcinoma is infrequently associated with the Merkel cell polyomavirus

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