Production of policlonal antisera specific to plant viruses by rabbit oral immunization



Document title: Production of policlonal antisera specific to plant viruses by rabbit oral immunization
Journal: Fitopatologia brasileira
Database: PERIÓDICA
System number: 000295132
ISSN: 0100-4158
Authors: 1

Institutions: 1Universidade Federal do Ceara, Departamento de Fitotecnia, Fortaleza, Ceara. Brasil
Year:
Season: Dic
Volumen: 26
Number: 4
Pages: 774-777
Country: Brasil
Language: Inglés
Document type: Artículo
Approach: Experimental
English abstract Serological techniques are of great importance for plant virus identification and characterization. The major limiting factor for using these techniques for plant virus identification is the requirement of a good virus purified preparation to be used in immunizing animals for antiserum production. In the present study, two New Zealand rabbits were orally immunized with extracts from cowpea (Vigna unguiculata) plants systemically infected with Cowpea severe mosaic virus (CPSMV) and with extracts from papaya (Carica papaya) infected with Papaya lethal yellowing virus (PLYV). The leaf extracts were prepared in saline solution 0.15 M in the rate of 1:1 (w/v) and clarified by a centrifugation of 10,000 g for 10 min. The clarified extracts containing the viruses were orally administered to the New Zealand rabbits in two series of five daily doses of 1.0 ml each. The obtained policlonal antisera were shown to be very specific to their respective viruses in double immunodiffusion and indirect ELISA. These seem to be the first antisera specific for plant virus obtained by rabbit oral immunization. The results open up some possibilities for producing antisera to plant viruses of difficult purification. It is a simple, fast and inexpensive method for production of antisera for plant viruses when compared to the traditional techniques that involve rabbit injections with purified virus preparations
Portuguese abstract As técnicas sorológicas são de grande importância na identificação de vírus, sendo que sua maior limitação na diagnose de viroses vegetais reside na dificuldade da purificação dos vírus para a produção de anti-soros. No presente trabalho, coelhos da raça Nova Zelândia Branco foram, individualmente, imunizados por via oral com extrato de plantas de caupi (Vigna unguiculata) infetadas pelo vírus do mosaico severo do caupi (Cowpea severe mosaic virus, CpSMV) e extrato de folhas de mamoeiro (Carica papaya) infetado com o vírus do amarelo letal (Papaya lethal yellowing, PLYV). Os extratos foram preparados em solução salina 0,15 M, na proporção de 1:1 (p/v), clarificados a uma centrifugação de 10.000 g por 10 min e administrados oralmente aos coelhos em duas séries de cinco doses diárias de 1,0 ml cada, com intervalo de dois dias entre cada série. Os anti-soros obtidos foram avaliados por difusão dupla em agar e ELISA indireto, tendo mostrado-se reativos com extratos de plantas infetadas, sem apresentar reação com extratos de plantas sadias. Embora com títulos baixos, os anti-soros apresentaram excelentes especificidades para extrato de plantas infetadas, criando perspectivas para produção de anti-soros contra vírus de plantas com boa imunogenicidade e de difícil purificação. Trata-se de uma técnica simples, rápida e barata para produção de anti-soro específico para vírus de plantas
Disciplines: Biología,
Agrociencias
Keyword: Inmunología,
Virus,
Leguminosas,
Serología,
Identificación,
Virus del mosaico del caupi
Keyword: Biology,
Agricultural sciences,
Immunology,
Virus,
Legumes,
Serology,
Identification,
Cowpea severe mosaic virus
Full text: Texto completo (Ver HTML)