In Situ Detection of PCR-amplified DNA and cDNA: A Review

Abstract

Because standard in situ hybridization protocols cannot routinely detect 1 to a few copies of DNA or RNA molecules, they are unable to detect latent viral infections, especially HIV or HPV viruses. This disadvantage has been the impetus for development of the polymerase chain reaction-in situ hybridization (PCR-ISH) method. The PCR-ISH can demonstrate early infections in tissue sections without the need for intact cells. It combines the sensitivity of the PCR with the cell localization of ISH, it has few false positive results because of sample contamination, and it provides information about the histologic distribution of the virus.

This paper reviews the strengths and weaknesses of the technique, its application in viral detection, and the techniques for achieving optimal results, especially with the use of the enzyme reverse transcriptase. (The J Histotechnol 17:235, 1994)