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

Medina, D. J.; Chang, P. W.; Bradley, Terence M.; Yeh, Min-Tsung

doi:10.3354/dao013147

Cited by 10 publications

(6 citation statements)

References 10 publications

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“…The greater of the mean absorbance of negative samples plus 3 standard deviations (SD) represent a cut‐off value with a 99% probability that a positive sample would yield a higher absorbance value (McLaren et al . ; Kemeny ; Medina et al ., ; MacPhee, et al ., ). Therefore, the negative–positive cut‐off OD point was established by adding the average OD of the negative samples to the SD multiplied by 3.…”

Section: Methodsmentioning

confidence: 99%

The development of an indirect ELISA for the detection of antibodies to goose parvovirus in blood serum

Fan

Zuo

Yang

et al. 2013

Lett Appl Microbiol

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Significance and Impact of the Study: In this study, we expressed and purified the capsid protein VP2 of GPV and developed a VP2-based i-ELISA for the detection of GPV antibodies. This i-ELISA can be used for evaluating the immune effects of GPV vaccine and for diagnosing of GPV infection. AbstractThis work aimed to develop an indirect enzyme-linked immunosorbent assay (i-ELISA), based on the structural protein VP2 of goose parvovirus (GPV), for the detection of antibodies to GPV in blood serum. In this study, the capsid protein VP2 of GPV was expressed in Escherichia coli and purified by nickel chromatography. The purified protein was capable of reacting with GPV antibodies, as analysed by Western blot. An i-ELISA based on the protein VP2 was developed. For evaluation of the newly developed ELISA, 192 field sera collected from six goose herds were tested in parallel by the serum neutralization (SN) test. The result showed that 138 samples were positive and 49 negative in both cases. The agreement of the two tests was 97Á3%. This i-ELISA has high sensitivity and good specificity and is an alternative serologic test for the detection of the antibodies to GPV in blood serum.

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Section: Methodsmentioning

confidence: 99%

The development of an indirect ELISA for the detection of antibodies to goose parvovirus in blood serum

Fan

Zuo

Yang

et al. 2013

Lett Appl Microbiol

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“…LaPatra et al (1989b) found that polyclonal and monoclonal antibody can be used to detect IHNV in blood and organ smears of juvenile salmonids and in ovarian fluid of adults. An ELISA assay detected as few as 70 PFU viruses in 100 µg of homogenized Atlantic salmon tissue and virus in cell culture media (Medina et al 1992). According to Arnzen et al (1991) the fluorescent antibody technique (FAT) is as sensitive as plaque assay and requires less time to obtain a confirmed diagnosis; however, monoclonal antibody against IHNV nucleoprotein and glycoprotein detected stages of virus replication in cell cultures as early as 6-8 hours postinoculation.…”

Section: Diagnosismentioning

confidence: 99%

Trout and Salmon Viruses

Plumb

Hanson

2010

Health Maintenance and Principal Microbial Diseases of Cultured Fishes

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“…ple would yield a higher absorbance value (McLaren et Comparison of the mean ELISA absorbance with the al. 1981, Kemeny 1991, Medina et al 1992. Analysis of concentration of bacteria in the graded concentration gill extracts from 34 normal fish (Tables 2 & 3), as well as bath challenge produced a linear relationship between from 126 normal fish tested In subsequent experiments exposure concentration and absorbance (Fig.…”

Section: Antigenic Specificitymentioning

confidence: 99%

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

Cited by 10 publications

References 10 publications

The development of an indirect ELISA for the detection of antibodies to goose parvovirus in blood serum

The development of an indirect ELISA for the detection of antibodies to goose parvovirus in blood serum

Trout and Salmon Viruses

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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