HPV detection in cytological cases with condylomatous or dysplastic changes: A study with PCR and in situ hybridization on cytological material

Abstract

Cytobrush samples of 80 patients, who previously had a cytological or histopathological diagnosis of condyloma and/or dysplasia were investigated for human papillomavirus infection (HPV) by polymerase chain reaction (PCR) and in situ DNA hybridization technique (ISH). The results were compared with concomitantly obtained cytological Pap-stained smears or, in some cases, histological sections. The time between the diagnosis of the original and the concomitant cytology/histopathology was less than 1 yr. Six additional patients had similar morphological diagnoses 2-4 yr before. Five more cases were included on clinical diagnosis of HPV. Compared with the original morphological diagnoses, 70% of the cases were positive by PCR and/or ISH. The concomitant morphology was not diagnostic of HPV in 44 out of 80 cases (55%), showing a relatively high percentage of cases morphologically normalized in the interval since the first specimen was taken. After detection with PCR, 30 cases (37.5%) were negative for HPV. Only one of the patients with a previous disease 2-4 yr before was HPV positive by PCR and two out of five patients with a clinical diagnosis of HPV. ISH could be performed on 67/80 cases, 43 of which were positive for HPV. There was a good agreement between the results of ISH and PCR, but there were six cases positive by ISH and negative by PCR. In these cases, few infected cells may have escaped detection by PCR. Both methods seem to be able to detect silent HPV infections and comparison with concomitant cytology/histopathology shows that morphology alone is insufficient for HPV detection in these cases.

HPV 16 infection in a patient with two primary squamous cell carcinomas: of the uterine cervix and the anal mucosa

“…HPV infections may regress, persist or progress, and the outcome cannot be predicted from the infected HPV type. Reports from our group and others have demonstrated HPV in up to 96% of cervical carcinomas (2,20,22,25,28,30) and 85% of anal carcinomas (17,26,29). HPV 16 was identified in most tumors of the cervix as well as of the anus, in agreement with the present findings.…”

“…Sections for immunohistochemistry were stained using the avidin-biotin-peroxidase complex (ABC) method (18). Deparaffinized sections were treated with 0.3'% hydrogen peroxide (H202) in methanol for 30 min to block endogenous peroxidase. To unmask the epitopes of p53, we microwaved the sections in 10 mM citrate buffer, pH 6.0, for 2 x 5 rnin (4).…”

Implications of human papillomavirus type for survival in cervical squamous cell carcinoma

“…HPV typing data on 75 of these cases have been published elsewhere (13). The present series includes 32 more cases, diagnosed in 1988.…”

Human papillomavirus type 16 capsids expose multiple type-restricted and type-common antigenic epitopes

“…This was performed in parallel with immunocytochemical staining to ascertain which HPV type each biopsy contained. ISH was performed either using biotin-labelled HPV probes (Vira Type in situ procedure, Life Technologies Inc; Biohit HPV in situ typing test, Biohit Oy) or with probes biotinylated by nick translation (Skyldberg et al, 1995). ISH with the two kits were performed according to the manufacturer's instructions, whereas the other hybridizations were performed essentially as described (Skyldberg et al, 1995).…”

“…ISH was performed either using biotin-labelled HPV probes (Vira Type in situ procedure, Life Technologies Inc; Biohit HPV in situ typing test, Biohit Oy) or with probes biotinylated by nick translation (Skyldberg et al, 1995). ISH with the two kits were performed according to the manufacturer's instructions, whereas the other hybridizations were performed essentially as described (Skyldberg et al, 1995). The tissue sections were treated before hybridization with 500 ktg/ml Proteinase K in phosphate-buffered saline (PBS), pH 7.2, for 15 min at 37 °C.…”