Antigen-capture ELISA for viral haemorrhagic septicaemia virus serotype I

Mourton, Chantal; Béarzotti, M.; Piechaczyk, Marc; Paolucci, F.; Pau, Bernard; Bastide, J.-M.; Kinkelin, P. de

doi:10.1016/0166-0934(90)90059-o

Cited by 27 publications

(22 citation statements)

References 7 publications

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“…15-mer peptides from the gpG of VHSV in solid phase were incubated with purified IgG isolated from murine ascites containing anti-gpG MAbs. Means and standard dev.iations from 2 different experiments are shown MAbs were I10 and C10 (Mourton, et al 1990), IPlH3 (Lorenzen et al 1990), lHlO (Sanz & Coll 1992, Sanz et al 1993) and 3FlA2 (Lorenzen unp'ubl.). MAbs 1H10, C10 and 3FlA2 were neutralizing.…”

Section: Discussionmentioning

confidence: 99%

“…nd: not determined. To obtain the % of neutralization shown in the table, MAbs C10 and 3FlA2 were used at 1 pg ml-l, MAb lHlO was used at 500 pg ml-l, MAb 1C3 was used at 65 p.g ml-' and MAb 4E6 was used at 10 pg mP1 The following MAbs had been described previously: I10 (Mourton et al 1990), IPlH3 (Lorenzen et al 1990), 1H10 (Sanz & Col1 1992), and C10 (Mourton et al 1990)…”

Section: Discussionmentioning

confidence: 99%

“…The concentrated samples were applied onto precoated nitrocellulose membrane strips (ImmunoType Kit, Sigma) to determine the MAbs isotypes by following the procedure recommended by the manufacturers of the kit. Other MAbs used were 3FlH10 and IPlH3 (Lorenzen et al 1990) and I10 kindly donated by Dr De Kinkelin (Mourton et al 1990). Neutralizing MAb C10 was obtained from Sanofi Diagnostic Pasteur (MarnesLa-Coquette, France).…”

Section: Methodsmentioning

confidence: 99%

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Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Fernandez‐Alonso¹,

Lorenzo²,

Pérez³

et al. 1998

Dis. Aquat. Org.

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Antibody linear epitopes of the glycoprotein G (gpG) of the viral haemorrhagic septicaemia virus (VHSV), a rhabdovirus of salrnonids, were mapped by pepscan using overlapping 15-mer peptides covering the entire gpG sequence and ELISA with polyclonal and monoclonal murine and polyclonal trout antibodies. Among the regions recognized in the pepscan by the polyclonal antibodies (PAbs) were the previously identified phosphatidylserine binding heptad-repeats (Estepa & Col1 1996; Virology 216:60-70) and leucocyte stimulating peptides (Lorenzo et al. 1995; Virology 212:348-355). Among 17 monoclonal antibodies (MAbs), only 2 non-neutralizing MAbs, 110 (aa 139-153) and IPlH3 (aa 399-413), could b e mapped to specific peptides in the pepscan of the gpG. Mapping of these MAbs was confirmed by immunoblotting with recombinant proteins and/or other synthetic peptides covering those sequences.None of the neutralizing MAbs tested reacted with any of the gpG peptides. Previously mapped MAb resistant mutants in the gpG did not coincide with any of the linear epitopes defined by the pepscan strategy, suggesting the complementarity of the 2 methods for the identification of antibody recogn~tion sites.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Fernandez‐Alonso¹,

Lorenzo²,

Pérez³

et al. 1998

Dis. Aquat. Org.

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“…Using Mab ELISA for cell culture fluids. These concentrations may in a similar ELISA, Mourton et al (1990) could detect be detected when less stringent criteria are used.…”

Section: Detection O F Cell Culture Produced Ihnv B Y Elisamentioning

confidence: 99%

Diagnosis of infectious hematopoietic necrosis virus in Atlantic salmon Salmo salar by enzyme-linked immunosorbent assay

Medina¹,

Chang²,

Bradley³

et al. 1992

Dis. Aquat. Org.

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“…The ELISA is simple, sensitive and rapid; it facilitates quantitation of antigen; and the use of microtitre plates allows for the testing of a large number of samples (Voller et al 1976, Voller & Bidwell 1986, Arshkoosh & Kaattari 1990, Kemeny 1991. The ELISA is widely used for the identification of other fish pathogens, including Aeromonas salmonicida (Smith 1981, Austin et al 1986, Adams & Thompson 1990, infectious pancreatic necrosis virus (Dixon & Hill 1983), piscine rhabdoviruses (Dixon & Hill 1984), Yersinia ruckerii (Austin et al 1986), Renibacterium salmoninarum (Pascho & Mulcahy 1987), viral hemorrhagic septicemia (Way & Dixon 1988, Mourton et al 1990, Sanz & Col1 1992, infectious hematopoietic necrosis virus (Way & Dixon 1988, Medina et al 1992, and Vibrio anguillarum (Romestand et al 1993). The numerous advantages and wide use of the assay in aquaculture prompted our attempt to develop an ELISA that could detect antigens of Flavobacteriun~ branchiophilum.…”

Section: Introductionmentioning

confidence: 99%

Development of an enzyme-linked immunosorbent assay (ELISA) to estimate the quantity of Flavobacterium branchiophilum on the gills of rainbow trout Oncorhynchus mykiss

Dd¹,

Ve²,

Js³

et al. 1995

Dis. Aquat. Org.

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An enzyme-linked immunosorbent assay (ELISA) was developed to estimate the quantity of Flavobacterium branchiophilum in crude gill extracts from rainbow trout Oncorhynchus mykiss following bath exposure to the bacterium. The assay utilized the avidin-biotin system and polyclonal antiserum raised against the LAB 4a strain of F: branchiophilum. The detection threshold was ca 1 X 10"acteria rnl-', and during routine use the mean intra-assay and inter-assay variations were 6.7 % and 8.l'%, respectively The ELISA absorbance (405 nm) was proportional to the amounl of F branrh~o-phjlum present (within a range of antigen concentration of 0 to 80 000 cells ml-') whether whole bacterial cell preparations, g111 preparations splked with bacterial cells or extracts of infected gills were tested. In a comparison of whole cell preparations derived from the type strain of F branchiophilum (Amencan Type Culture Collection 35035), the LAB 4a strain and other common gill isolates (4 Flavobacten'um sp., a Flexlbactersp. and Aeromonas hydrophila), the assay proved specific for F branchiophiluni antigen. Adaption for field-collected samples is feasible, but will require further examination of the antigenic specificity, and re-optimization of the tissue sample concentration if gills from other species are to be tested. The ELISA is an achievable means of estimating the quantity of F: branchiophilum on the gills of large numbers of fish, and represents an important tool for bacterial gill disease research.

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Antigen-capture ELISA for viral haemorrhagic septicaemia virus serotype I

Cited by 27 publications

References 7 publications

Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Mapping of linear antibody epitopes of the glycoprotein of VHSV, a salmonid rhabdovirus

Diagnosis of infectious hematopoietic necrosis virus in Atlantic salmon Salmo salar by enzyme-linked immunosorbent assay

Development of an enzyme-linked immunosorbent assay (ELISA) to estimate the quantity of Flavobacterium branchiophilum on the gills of rainbow trout Oncorhynchus mykiss

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