Original Article

Mod Pathol 2001;14(7):702–709

Signal-Amplified Colorimetric In Situ Hybridization for Assessment of Human Papillomavirus Infection in Cervical Lesions

DAKO reagents were provided by DAKO Austria; KREATCH reagents by Zamponi Diagnostics, Austria; Viroactiv system by Virofem Diagnostica, Germany.

Peter Birner M.D.1, Barbara Bachtiary M.D.2, Bettina Dreier Ph.D.2, Monika Schindl M.D.1, Elmar A Joura M.D.3, Gerhard Breitenecker M.D.1 and Georg Oberhuber M.D.1

  1. 1Institute of Clinical Pathology, Department of Gynecopathology and Cytology, University of Vienna, Austria
  2. 2Department of Radiotherapy and Radiobiology, University of Vienna, Austria
  3. 3Department of Gynecology and Obstetrics, University of Vienna, Austria

Correspondence: Dr. Peter Birner, University of Vienna, Institute of Clinical Pathology, Währinger Gürtel 18-20, A-1090 Vienna, Austria; e-mail: peter.birner@akh-wien.ac.at; fax: 43-1-405-3402.

Accepted 27 February 2001.

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Abstract

Detection and typing of human papillomavirus (HPV) infection may have a major impact in cervical-screening and follow-up. In this study various commercially available techniques for the detection of HPV were evaluated. HPV-status was determined in 86 samples of cervical cancer by PCR and direct sequencing, catalyzed signal amplified colorimetric DNA in situ hybridization (CSAC- ISH) (GenPoint system, DAKO), immunohistochemistry (IHC) and in 12 selected cases also by conventional, non-amplified ISH. Twenty-one samples of cervical intraepithelial neoplasias grade III (CIN III) were investigated by CSAC-ISH, conventional ISH and by IHC, in corresponding PAP smears HPV-detection and typing was performed by CSAC-ISH and Hybrid Capture test II (HC). In additional 20 PAP smears HPV typing was performed using HC and a novel immunocytochemical system for HPV detection and-typing. CSAC-ISH showed good correlation with PCR analysis in cervical cancers: In 87% of PCR positive cases, HPV infection was also detected by CSAC- ISH (66/76). HPV 16 was detected in 75% of PCR-positive cases (44/59), HPV 18 in 71% of PCR positive cases (5/7). CSAC-ISH detected HPV 31 in only 29% of PCR positive cases (2/7), and HPV 33 in 64% of PCR-positive cases (23/36). Nevertheless, CSAC-ISH- false negative cases for HPV 31 or 33 were nearly always combined infections with other HPV types, which were detectable by CSAC-ISH in most cases. CSAC-ISH revealed HPV infection in 20 of 21 HC-positive cervical smears, while in corresponding biopsies (CIN III) CSAC-ISH detected 100% of HPV infections. Conventional, non-amplified ISH showed significantly lower sensitivity compared with CSAC-ISH, and immunocyto- and -histochemistry were of very low sensitivity for detection of HPV. CSAC-ISH is an easy-to-handle method for detection and typing of cervical HPV infection, and shows sufficient sensitivity for clinical practice.

Keywords:

Human papillomavirus, cervical cancer, In situ hybridization, polymerase chain reaction, signal amplification, hybrid capture test

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