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A simple and rapid technique to process formalin-fixed,paraffin-embedded tissues for the detection of viruses by the polymerase chain reaction
Authors:Petra Kiene  Karin Milde-Langosch  Miriam Runkel  Kornelia Schulz  Thomas Löning
Affiliation:(1) Department of Pathology, University of Hamburg, Hamburg, Federal Republic of Germany;(2) Department of Gynaecological Pathology and Electron Microscopy, Clinics of Obstetrics and Gynaecology, University of Hamburg, Hamburg, Federal Republic of Germany;(3) Institut für Pathologie, Universitäts-Krankenhaus Eppendorf, Martinistraße 52, W-2000 Hamburg 20, Federal Republic of Germany
Abstract:Summary The use of chelating resin in a simple, rapid and efficient pre-treatment protocol to process formalin-fixed, paraffin-embedded specimens for the polymerase chain reaction (PCR) is described and compared to other pre-treatment techniques. With this modified PCR protocol, a variety of human autopsy and biopsy specimens were investigated for presence of DNA of human papil-lomaviruses, cytomegalovirus or Epstein-Barr virus. These viruses were detected in a productive or non-/low-productive state. Amplimers generated ranged from 88 to 450 base pairs length. Under the specified technical conditions, a considerable range of DNA molecules can be amplified from paraffin-embedded material, some older than 10 years.Dedicated to Prof. Dr. G. Seifert to his 70th birthday.
Keywords:Polymerase chain reaction  Human papillomavirus  Cytomegalovirus  Epstein-Barr virus
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