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Open Access Research article

Comparison of human papillomavirus detection between freshly frozen tissue and paraffin embedded tissue of invasive cervical cancer

Michael Odida12, Silvia de Sanjose34, Sven Sandin1, Beatriz Quiros3, Laia Alemany34, Belen Lloveras35, Wim Quint6, Bernhard Kleter6, Maria Alejo37, Leen-Jan van Doorn6 and Elisabete Weiderpass11089*

Author Affiliations

1 Department of Medical Epidemiology and Biostatistics, Karolinska Institutet, Stockholm, Sweden

2 Department of Pathology, Faculty of Medicine, Makerere University, Kampala, Uganda

3 Unit of Infections and Cancer, Cancer Epidemiology Research Programme (IDIBELL), Institut Català d' Oncologia, Barcelona, Spain

4 The Biomedical Research Centre Network for Epidemiology and Public Health (CIBERSP), Barcelona, Spain

5 Department of Pathology, Hospital del Mar, Barcelona, Spain

6 DDL Diagnostic Laboratory, Voorburg, The Netherlands

7 Department of Pathology, Hospital General de Vic, Barcelona, Spain

8 Department of Etiological Research, Cancer Registry of Norway, Oslo, Norway

9 Department of Community Medicine, Tromso University, Tromso, Norway

10 Department of Genetic Epidemiology, Folkhälsan Research Center, Helsinki, Finland

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Infectious Agents and Cancer 2010, 5:15  doi:10.1186/1750-9378-5-15

Published: 16 September 2010

Abstract

Background

Human Papillomavirus (HPV) detection results comparing paraffin embedded cervical tissue and other cervical specimens have been done with varying degrees of agreement. However, studies comparing freshly frozen specimens and paraffin embedded specimens of invasive cervical carcinomas are lacking. The aim of the study was to compare HPV detection using SPF10 broad-spectrum primers PCR followed by DEIA and genotyping by LiPA25 (version 1) between freshly frozen cervical tissue samples and paraffin embedded blocks of cervical tissue from the same patient. There were 171 pairs of paraffin embedded and freshly frozen samples analyzed from cervical carcinoma cases from Kampala, Uganda.

Results

88.9% (95% CI: 83.2%-93.2%) of paraffin embedded samples were HPV positive compared with 90.1% (95% CI: 84.6%-94.1%) of freshly frozen samples, giving an overall agreement in HPV detection between fresh tissue and paraffin embedded tissue at 86.0% (95% CI: 79.8%-90.8%). Although the proportion of HPV positive cases in freshly frozen tissue was higher than those in paraffin blocks, the difference was not statistically significant (p > 0.05). In both types of tissues, single HPV infections were predominant, with HPV16 accounting for 47% of positive cases. Comparison in the overall agreement, taking into accounts not only positivity in general, but also HPV types, showed a 65% agreement (complete agreement of 59.7%, partial agreement of 5.3%) and complete disagreement of 35.0%. HPV detection in squamous cell carcinomas (SCC) and adenocarcinomas (ADC) was similar in fresh tissue or paraffin blocks (p ≥ 0.05).

p16 immunostaining in samples that had at least one HPV negative results showed that 24 out of 25 cases had an over-expressed pattern.

Conclusions

HPV DNA detection was lower among ADC as compared to SCC. However, such differences were minimized when additional p16 testing was added, suggesting that the technical issues may largely explain the HPV negative cases.