RESUMO
Conserved coat protein region of plant viruses is often used as source of antigen for production of polyclonal antibodies for broad-based detection of closely related viruses. Antigenic region in coat protein is located either on N-terminal, and/or C-terminal or in the middle of coat protein. A study was undertaken to determine if antigenic region resides in N-terminal in Garlic virus X (GarV-X) of Allexivirus. In allexiviruses, N-terminal of coat protein region (1-57 amino acids) was highly variable. A complete coat protein of 27 kDa and a truncated protein without N-terminal (20 kDa) of GarV-X were expressed in pET expression vector and confirmed in western blotting using anti-His antisera. These expressed proteins were purified and used for antisera production. Specific and strong reaction was obtained for antisera generated against GarV-X full CP and GarV-X was detected in field-grown allium crops viz., onion, garlic, leek, and bunching onion and chives in ELISA. Antisera against GarV-X CPΔ1-61 (truncated CP) did not show reaction for GarV-X detection in immunoassay. Epitope mapping also indicated N-terminal as major antigenic determinant region with highest antigenic signal score. Our studies confirm that antigenic signals or epitopes reside in the N-terminal region of GarV-X which can be synthesized and used for production of monoclonal antibodies for specific detection purposes.
Assuntos
Proteínas do Capsídeo/análise , Proteínas do Capsídeo/imunologia , Flexiviridae/imunologia , Flexiviridae/isolamento & purificação , Doenças das Plantas/virologia , Antígenos Virais/análise , Antígenos Virais/genética , Antígenos Virais/imunologia , Proteínas do Capsídeo/genética , Mapeamento de Epitopos , Flexiviridae/genética , Alho/virologia , Imunoensaio , Dados de Sequência Molecular , Proteínas Mutantes/análise , Proteínas Mutantes/genética , Proteínas Mutantes/imunologia , Cebolas/virologia , RNA Viral/genética , Análise de Sequência de DNA , Testes SorológicosRESUMO
The presence of vertical ocular motor apraxia should prompt a careful assessment to detect an underlying neurometabolic condition. But congenital vertical ocular motor apraxia is a rare entity that may be attributed to perinatal stroke, hypoxia or kernicterus. We report a case of a 5-year old girl with congenital vertical ocular motor apraxia. Her magnetic resonance imaging (MRI) brain showed symmetrical high signal in the inferomedial thalamic nuclei in the distribution of the vessel of Percheron, a rare posterior cerebral artery variant. Awareness of this rare cause of childhood stroke may obviate unnecessary investigation.