Um ensaio RT-rtPCR para detecção de vírus da raiva em espécimes bovinos

Ciencia Rural

article

Gisane Lanes de Almeida¹  Francielle Liz Monteiro¹  Ingryd Merchioratto¹  Ana Paula Gnocato Mortari¹  Juliana Felipetto Cargnelutti¹  Rudi Weiblen¹  Eduardo Furtado Flores¹ 

[1] Universidade Federal de Santa Maria

关键词: rabies;    diagnostic;    cattle;    PCR;   

DOI  :  10.1590/0103-8478cr20210709

学科分类：农业科学（综合）

来源: Universidade Federal de Santa Maria * Centro de Ciencias Rurais

【 摘 要 】

Bovine rabies is endemic in most Brazilian States, including Rio Grande do Sul (RS), which has faced an unprecedented rabies outbreak between 2011 and 2018. We described a real-time reverse transcription quantitative PCR (RT-rtPCR) for detection of rabies virus (RABV) in bovine samples. The primers were designed targeting a highly conserved region of the nucleoprotein (N) gene of RABV obtained from cattle. The detection limit corresponded to 13 DNA copies and the intra- and inter-run repeatability was adequate (CV<9%) in all dilutions tested. Amplification of other pathogens associated with neurological disease in cattle or cross-contamination was not observed. Brain samples from cattle suspicious of rabies (n=21) were tested in triplicate by the RT-rtPCR and by the gold-standard direct fluorescent antibody test (DFAT), resulting in 100% of sensitivity and specificity of the RT-rtPCR. Testing of additional 41 bovine brain samples submitted to the routine DFAT testing yielded 37 (90.2%) concordant results (30 positive/7 negative) and 4 (9.7%) inconclusive in DFAT and RT-rtPCR positive. These results showed a good concordance between the tests and a higher sensitivity of the RT-rtPCR. This assay represents an alternative for RABV detection, either as a confirmatory test or for large-scale diagnosis in endemic regions.

【 授权许可】

Unknown