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Ultrasensitive detection of oncogenic human papillomavirus in oropharyngeal tissue swabs.
Isaac, Andre; Kostiuk, Morris; Zhang, Han; Lindsay, Cameron; Makki, Fawaz; O'Connell, Daniel A; Harris, Jeffrey R; Cote, David W J; Seikaly, Hadi; Biron, Vincent L.
Affiliation
  • Isaac A; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Kostiuk M; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Zhang H; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Lindsay C; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Makki F; , 1E4 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • O'Connell DA; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Harris JR; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Cote DW; , 1E4 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Seikaly H; Division of Otolaryngology-Head and Neck Surgery, Department of Surgery, University of Alberta, 1E4.34 WMC, 8440 112 Street, Edmonton, AB, T6G 2B7, Canada.
  • Biron VL; , 1E4.29 WMC 8440 112 Street, Edmonton, AB, T6G 2A1, Canada.
J Otolaryngol Head Neck Surg ; 46(1): 5, 2017 Jan 14.
Article in En | MEDLINE | ID: mdl-28088212
ABSTRACT

BACKGROUND:

The incidence of oropharyngeal squamous cell carcinoma (OPSCC) caused by oncogenic human papillomavirus (HPV) is rising worldwide. HPV-OPSCC is commonly diagnosed by RT-qPCR of HPV E6 and E7 oncoproteins or by p16 immunohistochemistry (IHC). Droplet digital PCR (ddPCR) has been recently reported as an ultra-sensitive and highly precise method of nucleic acid quantification for biomarker analysis. To validate the use of a minimally invasive assay for detection of oncogenic HPV based on oropharyngeal swabs using ddPCR. Secondary objectives were to compare the accuracy of ddPCR swabs to fresh tissue p16 IHC and RT-qPCR, and to compare the cost of ddPCR with p16 IHC.

METHODS:

We prospectively included patients with p16+ oral cavity/oropharyngeal cancer (OC/OPSCC), and two control groups p16- OC/OPSCC patients, and healthy controls undergoing tonsillectomy. All underwent an oropharyngeal swab with ddPCR for quantitative detection of E6 and E7 mRNA. Surgical specimens had p16 IHC performed. Agreement between ddPCR and p16 IHC was determined for patients with p16 positive and negative OC/OPSCC as well as for healthy control patients. The sensitivity and specificity of ddPCR of oropharyngeal swabs were calculated against p16 IHC for OPSCC.

RESULTS:

122 patients were included 36 patients with p16+OPSCC, 16 patients with p16-OPSCC, 4 patients with p16+OCSCC, 41 patients with p16-OCSCC, and 25 healthy controls. The sensitivity and specificity of ddPCR of oropharyngeal swabs against p16 IHC were 92 and 98% respectively, using 20-50 times less RNA than that required for conventional RT-qPCR. Overall agreement between ddPCR of tissue swabs and p16 of tumor tissue was high at ĸ = 0.826 [0.662-0.989].

CONCLUSION:

Oropharyngeal swabs analyzed by ddPCR is a quantitative, rapid, and effective method for minimally invasive oncogenic HPV detection. This assay represents the most sensitive and accurate mode of HPV detection in OPSCC without a tissue biopsy in the available literature.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Papillomaviridae / RNA, Viral / Carcinoma, Squamous Cell / Oropharyngeal Neoplasms / Polymerase Chain Reaction Type of study: Diagnostic_studies / Observational_studies / Risk_factors_studies Limits: Adult / Aged / Female / Humans / Male / Middle aged Language: En Journal: J Otolaryngol Head Neck Surg Journal subject: OTORRINOLARINGOLOGIA Year: 2017 Type: Article Affiliation country: Canada
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Papillomaviridae / RNA, Viral / Carcinoma, Squamous Cell / Oropharyngeal Neoplasms / Polymerase Chain Reaction Type of study: Diagnostic_studies / Observational_studies / Risk_factors_studies Limits: Adult / Aged / Female / Humans / Male / Middle aged Language: En Journal: J Otolaryngol Head Neck Surg Journal subject: OTORRINOLARINGOLOGIA Year: 2017 Type: Article Affiliation country: Canada