Detection and genetic characterization of samples compatible with antigenic variant 2 (AgV2) of rabies virus (RABLV) by real-time RT-PCR (RT-qPCR)

• Grant number: 17/18812-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2018
• End date: December 31, 2018
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Karin Corrêa Scheffer Ferreira
• Grantee: Yasmin Machado de Freitas
• Host Institution: Instituto Pasteur (IP). Coordenadoria de Controle de Doenças (CCD). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

The antemortem diagnosis of human rabies can be performed using a test for rabies virus-neutralizing antibodies and rabies antigen detection by direct immunofluorescence, and viral isolation. The accuracy and sensitivity of these traditional techniques show great variation in success. The Pasteur Institute, a national reference laboratory for the human rabies diagnosis, advocates for the diagnosis antemortem of rabies, a hemi-nested RT-PCR followed by nucleotide sequencing of rabies N and G genes. However, despite the accuracy of molecular biology methods in the identification and genetic characterization of the etiologic agent, these procedures are laborious and the results can be time consuming. In view of this, techniques such as real-time RT-PCR should be improved due to the speed and accuracy in obtaining results, both in the detection and in the genetic characterization of the rabies virus. The present study will standardize the real time RT-PCR methodology with samples of street RABV of genetic lineage compatible with dog variant antigen (AgV2), which will be analyzed by the Taqman® system (hydrolysis probe). (AU)