Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions

  • Fernando J Beltran Instituto de Zoonosis Luis Pasteur, Buenos Aires, Argentina
  • Federico Gury Dohmen Instituto de Zoonosis Luis Pasteur, Buenos Aires, Argentina
  • Horacio Del Pietro Servicio Nacional de Sanidad y Calidad Agroalimentaria, Buenos Aires, Argentina
  • Daniel M Cisterna Servicio de Neurovirosis, INEI-ANLIS Dr. Carlos G. Malbrán, Buenos Aires, Argentina
Keywords: rabies, molecular typing, decomposed samples

Abstract

Introduction: The exposure of nervous tissue samples to high temperatures affects the sensitivity of rabies virus diagnostic tests, causing degradation of the viral structure. This study evaluated reverse transcriptase polymerase chain reaction (RT-PCR) for the diagnosis and molecular characterization of brain tissue samples in an advanced state of decomposition and poorly conserved viral isolates by comparing it with routine diagnostic tests.

Methodology: A panel of three canine brain samples exposed to controlled decomposition for 7, 15, 30, and 120 days were evaluated using fluorescence antibody test (FAT), mouse inoculation test (MIT), and RT-PCR. In addition, 14 isolates of rabies variants, representing the largest circulation in Argentina, preserved in inadequate cooling for six to eight years were analyzed. Molecular typing of strains was performed using a 159-nucleotide region corresponding to the nucleoprotein gene.

Results: The three samples analyzed were positive by RT-PCR at all the decomposition times evaluated, in contrast to results observed with FAT and MIT, which rapidly became negative. In addition, 100% of the inadequately preserved samples were characterized molecularly. The limit of detection of RT-PCR was 0.5 MICDL50/0.03 mL.

Conclusion: RT-PCR can be useful for rabies diagnosis and typing of putrefying samples or rabies isolates stored in inadequate conditions.

Author Biography

Daniel M Cisterna, Servicio de Neurovirosis, INEI-ANLIS Dr. Carlos G. Malbrán, Buenos Aires, Argentina

Servicio de Neurovirosis, Instituto Nacional de Enfermedades Infecciosas

Published
2014-08-13
How to Cite
1.
Beltran FJ, Dohmen FG, Del Pietro H, Cisterna DM (2014) Diagnosis and molecular typing of rabies virus in samples stored in inadequate conditions. J Infect Dev Ctries 8:1016-1021. doi: 10.3855/jidc.4136
Section
Original Articles