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A comparison of clinically utilized human papillomavirus detection methods in head and neck cancer
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Journal Article
A comparison of clinically utilized human papillomavirus detection methods in head and neck cancer
2011
Overview
Detection of human papillomavirus (HPV) in head and neck cancer has therapeutic implications.
In situ
hybridization and immunohistochemistry for p16 are used by surgical pathologists. We compared the sensitivity and specificity of three popular commercial tests for HPV detection in head and neck squamous cell carcinomas with a ‘gold standard’ HPV PCR assay. A total of 110 prospectively collected, formalin-fixed tumor specimens were compiled onto tissue microarrays and tested for HPV DNA by
in situ
hybridization with two probe sets, a biotinylated probe for high-risk (HR) HPV types 16/18 (Dako, CA, USA) and a probe cocktail for 16/18, plus 10 additional HR types (Ventana, AZ, USA). The p16
INK4
expression was also assessed using a Pharmingen immunohistochemistry antibody (BD Biosciences, CA, USA). Tissue microarrays were stained and scored at expert laboratories. HPV DNA was detected by MY09/11-PCR, using Gold AmpliTaq and dot-blot hybridization on matched-fresh frozen specimens in a research laboratory. HPV 16
E6
and
E7
-RNA expression was also measured using RT-PCR. Test performance was assessed by a receiver operating characteristic analysis. HR-HPV DNA types 16, 18 and 35 were detected by MY-PCR in 28% of tumors, with the majority (97%) testing positive for type 16. Compared with MY-PCR, the sensitivity and specificity for HR-HPV DNA detection with Dako
in situ
hybridization was 21% (95% confidence interval (CI): 7–42) and 100% (95% CI: 93–100), respectively. Corresponding test results by Ventana
in situ
hybridization were 59% (95% CI: 39–78) and 58% (95% CI: 45–71), respectively. The p16 immunohistochemistry performed better overall than Dako (
P
=0.042) and Ventana (
P
=0.055), with a sensitivity of 52% (95% CI: 32–71) and specificity of 93% (95% CI: 84–98). Compared with a gold standard HPV-PCR assay, HPV detection by
in situ
hybridization was less accurate for head and neck squamous cell carcinoma on tissue microarrays than p16 immunohistochemistry. Further testing is warranted before these assays should be recommended for clinical HPV detection.
Publisher
Nature Publishing Group US,Elsevier Limited
Subject
/ Aged
/ Biomarkers, Tumor - analysis
/ Biopsy
/ Carcinoma, Squamous Cell - chemistry
/ Carcinoma, Squamous Cell - diagnosis
/ Carcinoma, Squamous Cell - virology
/ Cyclin-Dependent Kinase Inhibitor p16 - analysis
/ Female
/ Genetics
/ Head and Neck Neoplasms - chemistry
/ Head and Neck Neoplasms - diagnosis
/ Head and Neck Neoplasms - virology
/ Humans
/ Male
/ Medicine
/ Oncogene Proteins, Viral - genetics
/ Papillomavirus E7 Proteins - genetics
/ Papillomavirus Infections - complications
/ Papillomavirus Infections - diagnosis
/ Papillomavirus Infections - virology
/ Patients
/ Probes
/ Repressor Proteins - genetics
/ Tumors
