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Journal of Clinical Microbiology, August 2004, p. 3758-3765, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3758-3765.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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

Karin Biedermann,1 Nadia Dandachi,1,2 Maria Trattner,3 Georgia Vogl,1 Hildegard Doppelmayr,4 Elena Moré,4 Alfons Staudach,3 Otto Dietze,1 and Cornelia Hauser-Kronberger1*

Institute of Pathology,1 Institute of Gynecology, Alfons,3 Department of First Internal Medicine, Private Medical School, Salzburg,4 Division of Oncology, Department of Internal Medicine, Karl-Franzens University, Graz, Austria2

Received 14 November 2003/ Returned for modification 10 January 2004/ Accepted 14 March 2004

Archival paraffin-embedded tumor specimens offer a wealth of information for both cancer research and for routine clinical applications. However, the use of formalin-fixed, paraffin-embedded specimens for quantitative real-time PCR is not yet a standard diagnostic method in many laboratories, in particular for the quantification of human papillomavirus (HPV). Particularly high-risk HPV types are involved in almost 100% of the carcinogenesis of cervical cancer. We compared the diagnostic applicability and sensitivity of real-time PCR to that of chromogenic tyramide-signal-amplified in situ hybridization and conventional PCR for the detection of HPV from archival tissue in 164 cases of carcinoma in situ and cervical cancer. Furthermore, we examined whether the viral load of HPV is of prognostic relevance. Our findings indicate that patients in tumor stage I with a lower viral load of HPV type 16 (HPV16; up to 1,000 copies/ng of DNA) had a significantly better survival than HPV 16-negative patients (P = 0.037). We observed a greater sensitivity of both real-time PCR and conventional PCR for the detection of HPV16 and -18 compared to signal amplified in situ hybridization. We found a considerable concordance between HPV16 ({kappa} = 0.661) and HPV18 ({kappa} = 0.781) status as measured by real-time PCR and conventional PCR, indicating similar sensitivities. We recognized an inhibitory effect of formalin fixation and paraffin embedding on the evaluation of real-time PCR quantification.

* Corresponding author. Mailing address: Institute of Pathology, Landeskliniken Salzburg, Muellner Hauptstrasse 48, A-5020 Salzburg, Austria. Phone: 43-662-4482-4731. Fax: 43-4482-882. E-mail: c.hauser-kronberger{at}lks.at.

Journal of Clinical Microbiology, August 2004, p. 3758-3765, Vol. 42, No. 8
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.8.3758-3765.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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