In situ reverse transcription-polymerase chain reaction: A novel technique for detection of rabies virus RNA in murine neuroblastoma cells

摘要

Rabies is the most dreaded disease that poses continuous challenge to control measures adopted all over the world, especially in developing countries like India where human rabies is often a serious problem. Although the diagnosis of rabies has improved over the last few years, laboratory facilities still play a major role. Molecular techniques, especially polymerase chain reaction (PCR) finds its application to many areas of diagnostics and also provides an increased sensitivity with less labor and time. Use of non-radioactive labels such as biotin and digoxigenin minimize the costs and also meet the safety concerns. The objective of the present study was to assess usefulness of an in situ reverse transcription-PCR (ISRT-PCR) technique incorporating a digoxigen-labeled nucleotide into viral ribonucleoprotein as a means for identifying rabies virus antigen in murine neuroblastoma cells infected with street rabies virus isolates. The same has been compared for sensitivity with a direct fluorescent antibody test (FAT).