Investigation of a broad-spectrum PCR assay for human papillomaviruses in screening benign lesions of the upper aerodigestive tract.

BACKGROUND: A variety of different human papillomavirus (HPV) types can be found in benign and malignant lesions of the upper aerodigestive tract. Therefore a broad-spectrum assay is needed for screening reasons. METHODS: A PCR system with degenerate consensus primers originating from a very conserved region (e.g. L1) of the HPV genome was applied. The sensitivity level was improved by combining PCR and nested PCR. RESULTS: A total of 27 biopsies from laryngeal papillomas (9), exophytic (3) and inverted (6) papillomas of the nasal cavity or paranasal sinuses, papillomas of the uvula or soft palate (5), leukoplakias of the larynx (2), seborrheic keratosis (1) and granulation tumor of the tongue (1) were analyzed by the broad-spectrum PCR system. Sixteen cases showed a positive result in either PCR or nested PCR or both. CONCLUSIONS: It was shown that the applied broad-spectrum PCR system is a reliable tool in the detection of HPV DNA in benign lesions of the upper aerodigestive tract.

A two-tier polymerase chain reaction direct sequencing method for detecting and typing human papillomaviruses in pathological specimens.

An in-house polymerase chain reaction direct sequencing (PCR-DS) approach for HPV detection and typing was developed, taking advantage of two widely used pairs of human papillomavirus (HPV)-specific PCR primers, MY09/MY11 and GP5/GP6, and 33P-labeled dideoxynucleotides. In this study, 105 pathological specimens were examined: 89% were diagnosed as cervical intraepithelial neoplasia (CIN) grade I-III, 76.2% were HPV-positive by PCR-DS. The PCR using GP5/GP6 (first tier) and MY09/MY11 primers (second tier for the GP5/GP6-negative samples) detected additional 15%-25% HPV-positive samples compared with each pair used separately. Direct sequencing was then used to type the HPV. A readout of a sequence as short as 34 nucleotides within a specific region in the L1 gene is sufficient to type known or novel sequences. Because of its high sensitivity and cost-effectiveness, the two-tier PCR-DS was adopted by the authors as the current method of choice for HPV diagnosis with ultimate sequence precision.

Oral condyloma acuminatum associated with human papillomavirus and p53 overexpression mimicking cyclosporine effect in a transplant patient.

A case of human papillomavirus-associated condyloma acuminatum in the oral cavity, presumed to be fulminant cyclosporine-induced gingival hyperplasia, is reported in a 55-year-old cardiac transplant patient. Approximately 47 months following the transplant, the patient developed severe hyperplasia of the uvula and oral mucosa, resulting in difficulty swallowing. The histopathologic features of the lesion were typical of those of condyloma acuminatum. In situ hybridization of the paraffin-embedded material revealed infection with human papillomavirus types 6/11. This case lends further support to the putative role of long-term cellular immunosuppression in the development of human papillomavirus-associated squamous lesions. In addition, positive staining for p53 protein raises the possibility of concomitant p53 involvement in the pathogenesis of this oral lesion.