Virus isolation in cell culture for confirmatory diagnostic of rabies in bovine specimens

Abstract

This study investigated the suitability of virus isolation (VI) in mouse neuroblastoma cells (N2A) and baby hamster kidney cells (BHK-21) as a confirmatory test for diagnosis of bovine rabies. Fourty-eight brain samples from cattle suspected of rabies were initially submitted to fluorescent antibody test (FAT) and mouse inoculation test (MIT) for routine diagnostic. Subsequently, these specimens were submitted to three protocols of VI in each cell line: a single 24h or 72h passage (T1, T2), or three 48h passages (T3). The FAT and MIT combined detected 32/48 positive samples, from which MIT detected 32 and FAT 31. The average time required for final MIT results was 12.3 days (8 – 21). VI in BHK-21 cells provided definitive, positive results in 100% of the samples in 72h (T2) and in 96.9% after three 48h passages (T3). VI in N2A cells yielded positive results in 100% in 72h (T2) and in 93.7% of samples after three 48h passages (T3). Sensitivity, specificity, positive and negative predictive values were 100% in T2 in N2A and BHK-21 cells, and the Kappa value was excellent in both cells (k=1). A single 24h passage (T1) in both cell lines performed poorly, detecting less than 40% of the positive samples. Taking together, these results indicate that VI in both cell lines, especially in BHK-21 cells that grow faster and are much easier to maintain, does represent an adequate alternative for MIT as a confirmatory test for rabies diagnostic in bovine specimens, yielding reliable results in reduced time.