Use of a linear array for the detection of human papillomavirus genotypes in head and neck cancer.
BACKGROUND
Tumors of the head and neck commonly arise from the squamous and respiratory mucosa that lines the nasal and oral cavity, sinuses, pharynx, and larynx. The rate of oropharyngeal cancers diagnosed among Americans younger than 50 years is increasing. Infection of the oropharynx and tonsils by the human papillomavirus (HPV) has been linked to preneoplasia and cancer.


OBJECTIVE
To evaluate the Roche Linear Array HPV Genotyping test kit to identify, and then specifically genotype, HPV in formalin-fixed, paraffin-embedded tissues.


METHODS
We evaluated the performance of this assay for accuracy, for intra-assay and interassay precision, and for its limit of detection, using materials with known HPV status. Sixteen tumor tissues with the following origins were evaluated: 1 ocular, 1 hypopharynx, 8 tonsil, 1 retromolar trigone, 3 tongue, 1 anal, and 1 lymph node. DNA from formalin-fixed, paraffin-embedded tumor sections was isolated and amplified in duplicate, with positive and negative controls, using primers specific to the polymorphic L1 region of the HPV genome. Thirty-seven genotypes were tested using the linear array. The amplified product (450 base pairs) was visualized by gel electrophoresis and, if positive, reflexed to HPV genotyping.


RESULTS
Nine of the 16 tumors analyzed were HPV positive. The detected genotypes included HPV 6, 16, and 69.


CONCLUSIONS
The Roche Linear Array HPV Genotyping test is an easy-to-use method for determining HPV genotype in the routine analysis of formalin-fixed, paraffin-embedded tumors. This assay is robust and can be performed routinely in a clinical laboratory setting.
Department of Pathology and Medicine, Dartmouth Medical School, Dartmouth Hitchcock Medical Center, Lebanon, New Hampshire 03756, USA.
Candice C
CC
Black
Heather A
HA
Bentley
Thomas H
TH
Davis
Gregory J
GJ
Tsongalis
D000328:Adult; D000368:Aged; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005838:Genotype; D006258:Head and Neck Neoplasms / Q000473:pathology / Q000821:virology*; D006801:Humans; D057230:Limit of Detection; D008297:Male; D046228:Microarray Analysis; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results
D015344:DNA Probes, HPV; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic