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.
Department of Pathology, University Hospital, Uppsala, Sweden.
J F
JF
Sällström
I
I
Zehbe
M
M
Alemi
E
E
Wilander
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