Patterning and appliance of virus isolation techniques in mice neuroblastoma cells (N2A)
Keywords:
rabies, cell culture, murine neuroblastoma, batsAbstract
Rabies laboratorial diagnosis is very important since clinical diagnosis is not precise. Fluorescent Antibody Test (FAT) is the most used test and eventhough it is highly sensible, accurate and fast, false negatives results may occur. Thus, the isolation of rabies virus in mice (VIM) of Central Nervous System (CNS) samples of suspected animals to be infected is also recommended and, nowadays, this test has been substituted in many laboratories by viral isolation in cell culture (VICC). The aim of the present study was to compare the sensibility of virus isolation in murine neuroblastoma (N2A) cell culture with VIM test and with FAT, as well as evaluate obtained results in the diagnostic routine from Pasteur Institute, regarding reduction of costs, time and work. A total of 105 CNS samples of different animal species were analyzed by FAT, VIM and VICC: 50 bats, 32 dogs, 13 foxes and 10 bovines. All bats and bovines samples presented concordant results for the three tests, while dogs and foxes samples presented concordance only in 24 samples (69%).Based on these results, since 2004 it has been instituted that all bat samples sent to Pasteur Institute Laboratory, after diagnosed by FAT, should be submitted to viral isolation in cell culture, replacing the use of mice. In the period of January 2004 to September 2007, 11.298 bat samples were analyzed. A total of 67 positive samples for IFD and/or VICC were also submitted to VIM, and 61 samples presented concordant results for the three tests, and showed that the use of N2A cells is more sensible to “street virus” isolation of bat samples in laboratorial routine, being faster and lower in costs than VIM.
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King AA. Cell culture of rabies virus. In: Meslin F-X, Kaplan MM, Koprowski H. Laboratory techniques in rabies 1996. Geneva: World Health Organization. p. 114-130.
Webster WA, Casey GA. Virus isolation in neuroblastoma cell culture. In: Meslin F-X, Kaplan MM, Koprowski H. Laboratory techniques in rabies 1996 Geneva: World Health Organization. p. 96-104
Goldwasser RA, Kissling RE. Fluorescent antibody staining of street and fixed rabies vaccine antigens. Proc Soc Exp Biol Med. 1958; 98: 219-223.
Meslin F-X, Kaplan MM. An overview of laboratory techniques in the diagnosis and prevention of rabies and in rabies research. In: Meslin F-X, Kaplan MM, Koprowski H. Laboratory techniques in rabies 1996. Geneva: World Health Organization. p. 9-27.
Chhabra M, Mittal V, Jaiswal R, Malik S, Gupta M, Lal S. Development and evaluation of an in vitroisolation of street rabies virus in mouse neuroblastoma cells as compared to conventional tests used for diagnosis of rabies. Ind J of Med Microbiol. 2007; 25: 263-266.
Levaditi MC. Virus rabique et culture des cellules in vitro. C R Soc Biol. 1913; 75: 505.
Larghi OP, Nebel AE, Lazaro L, Savy VL. Sensitivity of BHK-21 cells supplemented with diethylamino ethyl-dextran for detection of street rabies virus in saliva simples. J Clin Microbiol.1975; 1: 243-245.
Smith AL, Tignor GH, Emmons RW, Woodie JD. Isolation of field rabies virus strains in CER and murine neuroblastoma cell cultures. Intervirology 1978; 9: 359-361.
Smith AL, Tignor GH, Mifini K, Motohaski T. Isolation and assay of rabies sero group viruses in CER cells. Intervirology 1977; 8: 92-99.
Rudd RJ e Trimarchi CV. Tissue culture technique for routine isolation of street strain rabies virus. J Clin Microbiol. 1987; 25: 1456-1458.
Tsiang H. An in vitro study of rabies pathogenesis. Bull Inst Pasteur 1985; 83: 41-56.
Tsiang H, Koulakoff A, Bizzini B, Berwald-Nether Y. J. Neuropathol Exp Neurol. 1983; 42: 439-452.
Umoh JU, Blenden DC. Comparasion of primary skunk brain and kidney and raccoon kidney cells with established cell lines for isolation and propagation of street rabies virus. Infect Immun.1983; 41: 1370-1372.
Dean DJ, Abelseth MK, Atanasiu P. Fluorescent antibody test. In: Meslin F-X, Kaplan MM, Koprowski H. Laboratory techniques in rabies 1996. Geneva: World Health Organization. p. 88-95
Koprowski, H. The mouse inoculation test. In: Meslin F-X, Kaplan MM, Koprowski H. Laboratory techniques in rabies 1996 Geneva; World Health Organization. p. 80-87.
Dean DJ, Abelseth MK. The fluorescent antibody test. In: Kaplan MM, Koprowski H. Laboratory techniques in rabies 1973 Geneva, World Health Organization. p. 73-83
Rudd RJ, Trimarchi CV. Development and Evaluation of an in vitro virus isolation procedure as a replacement for the mouse inoculation test in rabies diagnosis. J Clin Microbiol. 1989; 27: 2522-28.
Chhabra M, Bhardwaj M, Ichhpujani RL, Lal S. Comparative evaluation of commonly used laboratory test for post mortem diagnosis of rabies. Ind J Path Microbiol. 2005; 48: 190-93.
Chitra L, Pandit V, Kalyanaraman VR. Use of murine neuroblastoma culture in rapid diagnosis of rabies. Ind J Med Res. 1988; 87: 113-16.
Webster WA. A tissue-culture infection test in routine rabies diagnosis. Can J Vet Rec.1987; 87: 113-16.
Carrieri ML, Peixoto ZM, Paciência ML, Kotait I, Germano PM. Laboratory diagnosis ofequine rabies and its implications for human post exposure prophylaxis. J Virol Methods 2006; 138: 1-9.
Kotait I et al. Reservatórios silvestres do vírus da raiva: um desafio para a saúde pública. Boletim Epidemiológico Paulista 2007; 4: 1-8.
Kotait I. Programa de prevenção e controle da raiva transmitida por morcegos em áreas urbanas. Boletim Epidemiológico Paulista 2006; 3 (36): on line.
Trimarchi CV, Nadin Davis SA. Diagnostic Evaluation. In: Jackson AC, Wunner WH. Rabies.San Diego, CA, USA. 2ed. p. 411-462.
Carrieri et al. Desmodus rotunduscomo transmissor da raiva canina e feline no Estado de São Paulo. In: Seminário Internacional de Raiva, 2000 ago; São Paulo: Anais. p.42.
Carrieri ML, De Matos CA, Carnieli JrP, De Matos C, Favoretto SR, Kotait I. Canine and feline rabies transmitted by variant 3-Desmodus rotundus in the State of São Paulo, Brazil. In: Seminário Internacional: Morcegos como transmissores da raiva. 2001 dez; São Paulo: Anais.
p.51.
Kotait I, Carrieri ML. Raiva. In: Trabulsi LR, Alterthum F. Microbiologia. São Paulo: Atheneu, 2004. p. 651-57.
Peixoto ZMP, Cunha EMS, Sacramento D, Souza MCM, Queiroz da Silva LH, Germano PML, Kotait I. Rabies laboratory diagnosis: peculiar features of samples from equine origin. Braz J Microbiol. 2000; 31: 72-75.
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Copyright (c) 2007 Juliana Galera Castilho, Keila Iamamoto, Jonas Yoshitaka de Oliveira Lima, Karin Corrêa Scheffer, Pedro Carnieli Junior, Rafael de Novaes de Oliveira, Carla Isabel Macedo, Samira Maria Achkar, Maria Luiza Carrieri, Ivanete Kotait
This work is licensed under a Creative Commons Attribution 4.0 International License.
