Low prevalence of SV40 in Swiss mesothelioma patients after elimination of false-positive PCR results

Lung Cancer. 2007 Sep;57(3):282-91. Epub 2007 May 21. [Link]

Annemarie Zieglera, Christian A. Seemayerb, Marc Hinterbergerc, Peter Vogtc, Colette Bigoscha, Oliver Gautschid, Luigi Tornillob, Daniel C. Betticherd, Holger Mochc, Rolf A. Stahela

  • a Clinic and Policlinic of Oncology, Laboratory of Molecular Oncology, University Hospital, Haeldeliweg 4, CH-8044 Zurich, Switzerland
  • b Institute of Pathology, University Hospital, Basle, Switzerland
  • c Institute of Surgical Pathology, Department of Pathology, University Hospital, Zurich, Switzerland
  • d Institute of Medical Oncology, University Hospital, Berne, Switzerland

Received 1 December 2006; received in revised form 9 March 2007; accepted 22 March 2007


The association of simian virus 40 (SV40) with malignant pleural mesothelioma is currently under debate. In some malignancies of viral aetiology, viral DNA can be detected in the patients’ serum or plasma. To characterize the prevalence of SV40 in Swiss mesothelioma patients, we optimized a real-time PCR for quantitative detection of SV40 DNA in plasma, and used a monoclonal antibody for immunohistochemical detection of SV40 in mesothelioma tissue microarrays. Real-time PCR was linear over five orders of magnitude, and sensitive to a single gene copy. Repeat PCR determinations showed excellent reproducibility. However, SV40 status varied for independent DNA isolates of single samples. We noted that SV40 detection rates by PCR were drastically reduced by the implementation of strict room compartmentalization and decontamination procedures. Therefore, we systematically addressed common sources of contamination and found no cross-reactivity with DNA of other polyomaviruses. Contamination during PCR was rare and plasmid contamination was infrequent. SV40 DNA was reproducibly detected in only 4 of 78 (5.1%) plasma samples. SV40 DNA levels were low and not consistently observed in paired plasma and tumour samples from the same patient. Immunohistochemical analysis revealed a weak but reproducible SV40 staining in 16 of 341 (4.7%) mesotheliomas. Our data support the occurrence of non-reproducible SV40 PCR amplifications and underscore the importance of proper sample handling and analysis. SV40 DNA and protein were found at low prevalence (5%) in plasma and tumour tissue, respectively. This suggests that SV40 does not appear to play a major role in the development of mesothelioma.

Keywords: Mesothelioma, SV40 virus, Plasma DNA, Quantitative real-time PCR, Tissue array analysis