[Sequence analysis of PCR amplification products of old formalin fixed conisation tissue sections].
OBJECTIVE
Standard analytical techniques (DNA hybridisation methods) require high-quality DNA. In very small samples and in processed tissue (e.g. analysis of DNA in paraffin-embedded archival tissues) these methods fail due to their restricted sensitivity.


METHODS
In this study we used the considerably more sensitive PCR for analysis of HPV-16-e6/-e7 DNA in histologic cone biopsy sections. HPV-16-e6 and -e7 ORF were amplified, subcloned into M13mp18 and sequenced.


RESULTS
The sequence is identical to the published e6/e7 sequence.


CONCLUSIONS
Due to its high sensitivity the PCR is suitable to give answers to questions in cellular microbiology even in formalin-fixed paraffin-embedded tissues.
Labeit
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Universitäts-Frauenklinik Mannheim, Klinikum der Stadt Mannheim, Fakultät für Klinische Medizin, Universität Heidelberg.
Friese
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D002584:Cervix Uteri / Q000473:pathology; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D006801:Humans; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000235:genetics*; D050725:Papillomavirus E7 Proteins; D030361:Papillomavirus Infections / Q000473:pathology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012097:Repressor Proteins; D017422:Sequence Analysis, DNA; D014412:Tumor Virus Infections / Q000473:pathology*; D002583:Uterine Cervical Neoplasms / Q000473:pathology*
D004279:DNA, Viral; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16
Sequenzanalyse von PCR-Amplifikationsprodukten aus alten formalin- fixierten Konisationsschnittpräparaten.