Automated In situ Hybridization for Detection of Viral Pathogens Using RNAscope™.

In situ hybridization (ISH) is a powerful technique for detecting viral nucleic acids (viral RNA or DNA) within individual cells in tissue sections, allowing for the meaningful interpretation of the anatomical and histological localization of the target nucleic acids and providing relevant spatial context. The advent of novel ISH methods such as those utilizing a unique branched DNA "Z probe" coupled with an oligo-based signal amplification mechanism to amplify the signal from the RNA target (i.e., RNAscope™), have increased their sensitivity, as well as improved their reproducibility, allowing the detection of even low-abundance nucleic acid molecules in a single virus-infected cell. RNAscope™-based ISH has facilitated the detection of viral RNA and DNA with high sensitivity and specificity at single-cell resolution. This technique has significantly contributed to our understanding of viral pathogenesis, tropism, and host susceptibility for various human, animal, and plant viruses, and has also become a critical tool in establishing causality for new and/or emerging viral diseases.