In Situ PCR and Related Technology

Ever since the introduction of the polymerase chain reaction (peR) in 1986, morphologists, whose interests lie in the analysis of intact tissue structures, have been attempting to adapt this technique to intact cells or tissue sections to detect low copy numbers of DNA or RNA in situ while preserving tissue morphology. The significance of this objective is obvious. A technique finally materialized in 1990 when Dr. Ashley T. Haase and coworkers published results that used multiple prim ers with complementary tails in intact cells. Since then, a number of laboratories have successfully developed their own versions of the technique. In situ peR is now a well-recognized method that permits the detection of minute quantities of DNA or RNA in intact cells or tissue sections. As a result, morphological analysis of those target nucleotide sequences becomes possible. As anticipated, this ad vancement has led to significant improvement in our understanding of many nor mal and abnormal conditions, and its impact is becoming more evident as time passes. In situ peR has the characteristics of a new landmark in morphologic technol ogy-it is scientifically fascinating and technically challenging. In essence, it is a combination of in situ hybridization and conventional peR. The wealth of litera ture, experience and protocols for the two latter techniques can be applied to in situ peR. In situ peR also has its own unique aspects that were not addressed by the other two techniques.

Ever since the introduction of the polymerase chain reaction (peR) in 1986, morphologists, whose interests lie in the analysis of intact tissue structures, have been attempting to adapt this technique to intact cells or tissue sections to detect low copy numbers of DNA or RNA in situ while preserving tissue morphology.

In Situ PCR-An Overview.- In Situ PCR: General Methodology and Recent Advances.- Applications of In Situ PCR Methods in Molecular Biology.- In Situ PCR for the Detection of Human Papillomavirus in Cells and Tissue Sections.- Conventional PCR, In Situ PCR and Reverse Transcription In Situ PCR for HIV Detection.- Oligonucleotide-Primed In Situ Transcription and Immunogold-Silver Staining Systems: Localization of mRNAs in Tissues and Cells.- Sensitive Detection of DNA and mRNA Sequences by In Situ Hybridization and Immunogold-Silver Staining.- In Situ PCR: New Frontier for Histopathologists.- Color Illustrations.