title: [In situ PCR and PCR in situ hybridization of paraffin-embedded tissue. New diagnostic possibilities in pathology].
abstract: In situ polymerase chain reaction (PCR) and PCR in situ hybridization are new diagnostic tools in dermatopathology. These techniques can combine the sensitivity of PCR with the advantage of in situ hybridization to localize specific cellular structures. With optical control of the PCR reaction product in these techniques, false-positive results due to contamination can be minimized. Several working protocols have been established which allow detection of DNA and RNA sequences in a rapid and reproducible manner. Two different methods have been established to detect the amplification product, the indirect PCR in situ hybridization (PCRisHyb), and the direct in situ PCR (isPCR). An easy and reproducible method for PCRisHyb and isPCR in formalin-fixed and paraffin-embedded tissue is described and the two techniques are compared. Using both isPCR and PCRisHyb, the amplification product can be visualized with an optimal morphological preservation of the tissue. Indirect PCRisHyb showed a slightly higher specificity whereas direct isPCR was the quicker, easier and less expensive method.
authors.affiliation: Dermatologische Klinik, Ludwig-Maximilians Universität München.
authors.forename: P I
authors.initials: PI
authors.lastname: Schiller
authors.forename: U
authors.initials: U
authors.lastname: Puchta
authors.forename: A J
authors.initials: AJ
authors.lastname: Ogilvie
authors.forename: A
authors.initials: A
authors.lastname: Graf
authors.forename: P
authors.initials: P
authors.lastname: Kind
authors.forename: C A
authors.initials: CA
authors.lastname: Sander
mesh_terms: D002644:Chickenpox / Q000473:pathology; D006801:Humans; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D011237:Predictive Value of Tests; D020401:Primed In Situ Labeling / Q000295:instrumentation*; D012867:Skin / Q000473:pathology; D017193:Skin Diseases, Viral / Q000473:pathology