title: RNAscope for in situ detection of transcriptionally active human papillomavirus in head and neck squamous cell carcinoma.
abstract: The 'gold standard' for oncogenic HPV detection is the demonstration of transcriptionally active high-risk HPV in tumor tissue. However, detection of E6/E7 mRNA by quantitative reverse transcription polymerase chain reaction (qRT-PCR) requires RNA extraction which destroys the tumor tissue context critical for morphological correlation and has been difficult to be adopted in routine clinical practice. Our recently developed RNA in situ hybridization technology, RNAscope, permits direct visualization of RNA in formalin-fixed, paraffin-embedded (FFPE) tissue with single molecule sensitivity and single cell resolution, which enables highly sensitive and specific in situ analysis of any RNA biomarker in routine clinical specimens. The RNAscope HPV assay was designed to detect the E6/E7 mRNA of seven high-risk HPV genotypes (HPV16, 18, 31, 33, 35, 52, and 58) using a pool of genotype-specific probes. It has demonstrated excellent sensitivity and specificity against the current 'gold standard' method of detecting E6/E7 mRNA by qRT-PCR. HPV status determined by RNAscope is strongly prognostic of clinical outcome in oropharyngeal cancer patients.
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Wang
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Mindy Xiao-Ming
authors.initials: MX
authors.lastname: Wang
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Nan
authors.initials: N
authors.lastname: Su
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Li-Chong
authors.initials: LC
authors.lastname: Wang
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Xingyong
authors.initials: X
authors.lastname: Wu
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Son
authors.initials: S
authors.lastname: Bui
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Allissa
authors.initials: A
authors.lastname: Nielsen
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Hong-Thuy
authors.initials: HT
authors.lastname: Vo
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Nina
authors.initials: N
authors.lastname: Nguyen
authors.affiliation: Advanced Cell Diagnostics, Inc.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Advanced Cell Diagnostics, Inc.; xma@acdbio.com.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Ma
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D002294:Carcinoma, Squamous Cell / Q000821:virology*; D006258:Head and Neck Neoplasms / Q000821:virology*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000821:virology*; D012333:RNA, Messenger / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis; D000077195:Squamous Cell Carcinoma of Head and Neck
chemical_list: D014408:Biomarkers, Tumor; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D012367:RNA, Viral