Abstract:Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal … Show more
“…The T3 protocol (three passages of 48h each) failed in detecting one positive sample in BHK-21 cells and two in N2A. Previous studies have suggested a minimal incubation period of three (ANTÚNEZ et al, 2013) or 4-5 days (RUDD & TRIMARCHI, 1989), while others suggested that a 24h period would be adequate (BOURHY et al, 1989;CHITRA et al, 1988). These discrepancies may reflect different VI protocols or cells (cell lines, passage number, etc.…”
Section: ---------------Vi 1 In N2a Cells/protocol ------------------mentioning
confidence: 97%
“…Hence, virus isolation (VI) in cell culture has been proposed as an alternative method to replace MIT as a confirmatory test for rabies. Isolation systems based on mouse neuroblastoma (N2A) and baby hamster kidney (BHK-21) cells have been evaluated regarding to sensitivity to RabV from different animal species (RUDD et al, 1980;1989;RUDD & TRIMARCHI, 1987;ANTÚNEZ et al, 2013). Considering the fastidious growth and the special culture medium requirements for N2A cells, the use of BHK-21 cells for VI as confirmatory rabies diagnosis would compare favorably since BHK-21 cells display a fast growth in vitro and are largely used in many laboratories.…”
Section: Subsequentemente Essas Amostras Foram Submetidas a Três Promentioning
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
RESUMO
Este estudo investigou a sensibilidade do isolamento viral (VI) em células de neuroblastoma murino (N2A) e células de rim de hamster (BHK-21) como um teste confirmatório para o diagnóstico de raiva bovina. Quarenta e oito amostras de cérebro de bovinos com suspeita de raiva foram inicialmente submetidas aos testes diagnósticos de rotina, imunofluorescência direta (FAT) e inoculação intracerebral em camundongos (MIT).
Subsequentemente, essas amostras foram submetidas a três protocolos de VI em cada linhagem celular: uma única passagem de 24h ou 72h (T1, T2), ou três passagens de 48h (T3). Os testes FAT e MIT combinados detectaram 32/48 amostras positivas, das quais o MIT detectou 32 e a FAT 31. O tempo médio requerido para o resultado final no MIT foi 12,3 dias (8-21 dias). O teste de VI em células BHK-21 obteve resultados positivos em 100% das amostras em 72h (T2), e em 96,9% após três passagens de 48h (T3
“…The T3 protocol (three passages of 48h each) failed in detecting one positive sample in BHK-21 cells and two in N2A. Previous studies have suggested a minimal incubation period of three (ANTÚNEZ et al, 2013) or 4-5 days (RUDD & TRIMARCHI, 1989), while others suggested that a 24h period would be adequate (BOURHY et al, 1989;CHITRA et al, 1988). These discrepancies may reflect different VI protocols or cells (cell lines, passage number, etc.…”
Section: ---------------Vi 1 In N2a Cells/protocol ------------------mentioning
confidence: 97%
“…Hence, virus isolation (VI) in cell culture has been proposed as an alternative method to replace MIT as a confirmatory test for rabies. Isolation systems based on mouse neuroblastoma (N2A) and baby hamster kidney (BHK-21) cells have been evaluated regarding to sensitivity to RabV from different animal species (RUDD et al, 1980;1989;RUDD & TRIMARCHI, 1987;ANTÚNEZ et al, 2013). Considering the fastidious growth and the special culture medium requirements for N2A cells, the use of BHK-21 cells for VI as confirmatory rabies diagnosis would compare favorably since BHK-21 cells display a fast growth in vitro and are largely used in many laboratories.…”
Section: Subsequentemente Essas Amostras Foram Submetidas a Três Promentioning
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
RESUMO
Este estudo investigou a sensibilidade do isolamento viral (VI) em células de neuroblastoma murino (N2A) e células de rim de hamster (BHK-21) como um teste confirmatório para o diagnóstico de raiva bovina. Quarenta e oito amostras de cérebro de bovinos com suspeita de raiva foram inicialmente submetidas aos testes diagnósticos de rotina, imunofluorescência direta (FAT) e inoculação intracerebral em camundongos (MIT).
Subsequentemente, essas amostras foram submetidas a três protocolos de VI em cada linhagem celular: uma única passagem de 24h ou 72h (T1, T2), ou três passagens de 48h (T3). Os testes FAT e MIT combinados detectaram 32/48 amostras positivas, das quais o MIT detectou 32 e a FAT 31. O tempo médio requerido para o resultado final no MIT foi 12,3 dias (8-21 dias). O teste de VI em células BHK-21 obteve resultados positivos em 100% das amostras em 72h (T2), e em 96,9% após três passagens de 48h (T3
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