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Cell Culture Medium Inhibits Antigen Binding Used in an ELISA for Detection of Antibodies against Nervous Necrosis Virus.
Choi, Bomi; Gye, Hyun Jung; Oh, Myung-Joo; Nishizawa, Toyohiko.
Afiliación
  • Choi B; a Department of Aqualife Medicine , Chonnam National University , Yeosu 550-749 , South Korea.
J Aquat Anim Health ; 26(3): 168-72, 2014 Sep.
Article en En | MEDLINE | ID: mdl-25229488
ABSTRACT
Abstract We investigated the optimum dilution of nervous necrosis virus (NNV) for use as antigens to detect antibodies by an enzyme-linked immunosorbent assay (ELISA) in the Sevenband Grouper Epinephelus septemfasciatus. The ELISA values for a standardized suspension of antigens diluted with L-15 medium containing 1% fetal bovine serum decreased gradually with the dilution of the antigens, whereas those for the antigens diluted with distilled water (DW) initially increased with the dilution of the antigens, peaked at a 320-fold dilution, and then decreased thereafter. Additional studies revealed that binding of NNV antigens to ELISA wells was inhibited by fetal bovine serum and other substances in the L-15 medium. Sera obtained from Sevenband Grouper vaccinated with live NNV vaccine and survivors from natural NNV-infection were subjected to antibody detection by ELISA. All of the sera were positive by ELISA when the standardized suspension was diluted 320-fold, whereas sera from five out of the six survivors and two out of the six vaccinated fish were negative or weakly positive by ELISA using NNV antigens diluted 10-fold. We therefore concluded that cultured NNV solutions prepared in cell culture media may need to be diluted with distilled water for use in ELISA. Received January 28, 2014; accepted April 16, 2014.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Virus ARN / Lubina / Medios de Cultivo / Nodaviridae / Enfermedades de los Peces Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: J Aquat Anim Health Asunto de la revista: MEDICINA VETERINARIA Año: 2014 Tipo del documento: Article País de afiliación: Corea del Sur

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Infecciones por Virus ARN / Lubina / Medios de Cultivo / Nodaviridae / Enfermedades de los Peces Tipo de estudio: Diagnostic_studies Límite: Animals Idioma: En Revista: J Aquat Anim Health Asunto de la revista: MEDICINA VETERINARIA Año: 2014 Tipo del documento: Article País de afiliación: Corea del Sur
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