Pitfalls of in situ polymerase chain reaction (PCR) using direct incorporation of labelled nucleotides.
False positivity is reported of in situ PCR reactions in a direct incorporation assay with digoxigenin-labelled dUTP. It is recommended that in situ hybridization with specific labelled probe replaces the direct incorporation method for the detection of in situ PCR amplicon.
Sällström
J F
JF
Department of Pathology, University Hospital, Uppsala, Sweden.
Zehbe
I
I
Alemi
M
M
Wilander
E
E
D017209:Apoptosis; D001706:Biopsy; D002478:Cells, Cultured; D004247:DNA; D004279:DNA, Viral / Q000032:analysis; D004259:DNA-Directed DNA Polymerase; D003856:Deoxyuracil Nucleotides; D004076:Digoxigenin; D005189:False Positive Reactions; D005347:Fibroblasts / Q000166:cytology; D006561:Herpes Simplex / Q000175:diagnosis; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008198:Lymph Nodes / Q000473:pathology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011505:Protein-Tyrosine Kinases / Q000032:analysis; D011518:Proto-Oncogene Proteins / Q000032:analysis; D019253:Proto-Oncogene Proteins c-bcl-2; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification; D019914:Taq Polymerase; D014178:Translocation, Genetic
D004279:DNA, Viral; D003856:Deoxyuracil Nucleotides; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; C027078:deoxyuridine triphosphate; D004247:DNA; D011505:Protein-Tyrosine Kinases; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase; D004076:Digoxigenin