Abstract
Pyrosequencing is an alternative sequencing technique of small DNA fragments based on the sequencing-by-synthesis principle. Compared to the Sanger method, pyrosequencing is less costly and time consuming, although the applications of this technique do not completely overlap with those of the conventional method. In the last decade, pyrosequencing technology has been applied successfully to the typing of a number of pathogens, including lyssaviruses. An efficient protocol to lyssavirus detection and typing, by targeting the hypervariable region of the rabies nucleoprotein (N) gene, has been fully described in Chapter 20. A further protocol for the rapid identification of vaccine-associated rabies cases from field brain samples, targeting the viral glycoprotein (G) gene, is presented herein. Such a protocol could be used in association with the lyssavirus-typing protocol previously mentioned, and is a valid and sensitive alternative to monoclonal antibody (MAb) typing within the framework of a post-vaccination survey.
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