CARACTERIZACIÓN MOLECULAR DEL VIRUS DE LA RABIA EN EL ESTADO DE MEXICO

Abstract

Rabies virus is the most widespread and epidemiologically important member of the genus and the only taxon documented in America. Several specific Rabies virus variants have been characterized from different mammalian hosts, such as dogs, foxes, coyotes, raccoons, skunks, and multiple species of frugivorous, insectivorous, and hematophagous bats. Nowadays, many molecular studies have been performed targeting of the N gene, this data have permitted insights to virusreservoir relationships, patterns of transmission and dissemination, as well as viral evolution. Brain samples were using in this study tested positive by FAT and Monoclonal variants (INDRE) collected in Mexican State. hnRT-PCR and RT-PCR Syber green detection were done with own primer design in complete sequence of N gene rabies virus (genomic databases NCBI) concededly dog, skunk, non hemetophage bat and vampire bat as host. To confirm detection PCR products were sequenced and identificated in BLAST of the NCBI database. All of the brain samples from rabid animals, diagnosed as positive in the FAT test, signaled positive in the nRT-PCR and RT-PCR SYBR Green. To confirm the specificity of the both methods, they were carried out with the additional genotype 1 prototype laboratory strain CVS, used as a positive control. Real-time RT-PCR used primers of internal semi-nested end-point RT-PCR.In the characterization of the antigenic variants (AgV) with MABs in the dog sample, the dog primers identified the Variant dog (AgV1). This result matched with both the nRT-PCR and SYBR Green primers a 100%. Similarly, the skunk samples matched the same percentage with the skunk primers. On the other hand, in the bovine samples where the MABs detection identified the skunk Variant (AgV8), the determined host by nRT-PCR and SYBR Green diagnosed as positive with the vampire primers. The last two samples, the one determined as atypical and the one not determined with the MABs, the nRT-PCR and SYBR Green diagnosed as positive with the bat primers; proving thus the specificity of the RV variant detection depending on the host with he present study proposal. nRT-PCR and SYBR Green detection matched a 100% with the FAT and 80% with the MABs results.