Comparison between RT-PCR and the mouse inoculation test for detection of rabies virus in samples kept for long periods under different conditions.
Antigenic and genetic analyses are important tools for retrospective studies of rabies epidemiology in specific geographical areas. Virus recovery and re-isolation from archival samples conserved for long periods at freezing temperature are essential for these studies. Prolonged preservation, associated with temperature variations, causes significant loss of virus viability. However, molecular tools, such as RT-PCR, can overcome this problem. For this purpose, 95 positive samples stored for 4-13 years at -20 and -80 degrees C were evaluated by mouse inoculation test and RT-PCR. Only 32 (33.6%) of the samples were positive with the mouse inoculation test, while RT-PCR detected the viral genome in 62 (65.3%) samples. When the samples were analyzed in relation to storage period, there was a significant difference in those stored for >10 years, with 59.7% positivity for RT-PCR and 22.1% for mouse inoculation test. The present study confirms the significance of RT-PCR for detection of viral genomes in archival samples, including those in an apparent state of decomposition.
Lopes
Marissol Cardoso
MC
UNESP - Univ Est Paulista, Veterinary Medicine, Araçatuba Campus, Production and Animal Health Department, Rua Clóvis Pestana, 793 CEP 16050-680, Araçatuba, São Paulo, Brazil. marissolcl@hotmail.com
Venditti
Leandro Lima Rossignolo
LL
Queiroz
Luzia Helena
LH
D032761:Animal Experimentation; D000818:Animals; D006801:Humans; D051379:Mice; D050296:Microbial Viability; D011820:Rabies virus / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors