Detection of specific human papillomavirus types in paraffin‐embedded sections of cervical carcinomas

Human papillomaviruses (HPV) are the causative agents of most cervical carcinomas. A complete understanding of the HPV types that cause cervical carcinoma is needed as vaccines are designed. Fresh tissues are not always available for such studies. We therefore sought to determine the feasibility of HPV studies using formalin‐fixed, paraffin‐embedded sections of 56 cervical carcinomas, correlating typing information with the pathology and physical state of the HPV sequences within cells. Sections from each specimen were used to extract and purify DNA. Specific HPV types were identified using a PCR/reverse blot strip assay. Tyramide signal‐amplified, fluorescent DNA in situ hybridization (FISH) was used to localize HPV within cells. Human β‐globin sequences were amplified in DNA from all specimens. HPV sequences from oncogenic types were identified in 52 of 56 (92.9%) by PCR/reverse blot strip assay, and in one additional case using an HPV 16 multiplex PCR assay. HPV 16 was the most commonly detected type, present in most cases as a solitary isolate. Thirty‐ five of 42 HPV 16 or HPV 18 PCR‐positive specimens were also positive in the FISH assay, in most cases in a pattern consistent with viral integration. We conclude that HPV typing from formalin‐fixed, paraffin‐embedded sections of cervical carcinomas is possible, with a sensitivity that is similar to that found in studies using fresh tissue. J. Med. Virol. 78:117–124, 2006. © 2005 Wiley‐Liss, inc.

[1]  Julian Peto,et al.  Prevalence of Human Papillomavirus in Cervical Cancer: a Worldwide Perspective , 1995 .

[2]  D. Lowy,et al.  Minor capsid protein of human genital papillomaviruses contains subdominant, cross-neutralizing epitopes. , 2000, Virology.

[3]  B. Katz,et al.  Detection of multiple human papillomavirus types in condylomata acuminata from immunosuppressed patients. , 1994, The Journal of infectious diseases.

[4]  M. Schindl,et al.  Signal-Amplified Colorimetric In Situ Hybridization for Assessment of Human Papillomavirus Infection in Cervical Lesions , 2001, Modern Pathology.

[5]  R. Lloyd,et al.  In Situ Hybridization Detection of Low Copy Nucleic Acid Sequences Using Catalyzed Reporter Deposition and Its Usefulness in Clinical Human Papillomavirus Typing , 1998, Diagnostic molecular pathology : the American journal of surgical pathology, part B.

[6]  E. Vollmer,et al.  Comparison of in-situ hybridization, direct and indirect in-situ PCR as well as tyramide signal amplification for the detection of HPV , 1999, Histochemistry and Cell Biology.

[7]  M. Stoler,et al.  Detection of Multiple Human Papillomavirus Types in Condylomata Acuminata Lesions from Otherwise Healthy and Immunosuppressed Patients , 1999, Journal of Clinical Microbiology.

[8]  K. Cooper,et al.  Biotinyl-Tyramide-Based In Situ Hybridization Signal Patterns Distinguish Human Papillomavirus Type and Grade of Cervical Intraepithelial Neoplasia , 2002, Modern Pathology.

[9]  K. Cooper,et al.  Optimization of biotinyl-tyramide-based in situ hybridization for sensitive background-free applications on formalin-fixed, paraffin-embedded tissue specimens , 2003, BMC clinical pathology.

[10]  J. Peto,et al.  Human papillomavirus is a necessary cause of invasive cervical cancer worldwide , 1999, The Journal of pathology.

[11]  C. Gay,et al.  Human papillomavirus detection by non isotopic in situ hybridization, in situ hybridization with signal amplification and in situ polymerase chain reaction. , 1999, European journal of histochemistry : EJH.

[12]  H. Hausen Viruses In Human Tumors–Reminiscences And Perspectives , 1996 .

[13]  Daisy R. Lee,et al.  Detection of Human Papillomavirus in Archival Tissues: Comparison of In Situ Hybridization and Polymerase Chain Reaction , 1998, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[14]  David R. Scott,et al.  Improved Amplification of Genital Human Papillomaviruses , 2000, Journal of Clinical Microbiology.

[15]  R. Schlegel,et al.  Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. , 1985, The American journal of pathology.

[16]  A. Staudach,et al.  Comparison of Real-Time PCR Signal-Amplified In Situ Hybridization and Conventional PCR for Detection and Quantification of Human Papillomavirus in Archival Cervical Cancer Tissue , 2004, Journal of Clinical Microbiology.

[17]  K. Cooper,et al.  Episomal and integrated human papillomavirus in cervical neoplasia shown by non-isotopic in situ hybridisation. , 1991, Journal of clinical pathology.