Demonstration of Rabies Viral Nucleic Acids by In-Situ Polymerase Chain Reaction

This chapter introduces the development of the polymerase chain reaction (PCR) and its modifications that enabled the development of in-situ PCR and its successful application in localizing RNA and DNA targets. We provide detailed descriptions of methods and materials for successful demonstration of rabies virus (RABV) nucleic acid by in-situ PCR. The essential components of this topic include sample preparation, including care during pre-treatment, in-situ reverse transcription (RT) PCR amplification, and detection of target genes. The parameters pertaining to critical steps in the procedure are discussed.