Gel-Based Reverse Transcription-Polymerase Chain Reaction
The reverse transcription-polymerase chain reaction (RT-PCR) has been extensively applied, e.g. for epidemiological surveys and for ante-mortem diagnosis in humans. The conventional RT-PCR described here is one of the most widely used and has been validated to detect all recognized and putative lyssavirus species known to exist to date. This RT-PCR has also been widely used as a first amplification for sequence analysis of lyssaviruses. Despite the high level of sensitivity, the use of molecular assays, including RT-PCRs, for routine post-mortem diagnosis of lyssaviruses is currently not recommended by international organizations if brain tissue is available. However, if standardization, very stringent quality control, and sufficient experience and expertise are granted, the level of false results is minimal.