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Biomedical subjects

J Groen

Publications and source records attributed to J Groen.

At least 109 records · Page 6Linked to original sources

Mass mortality in seals caused by a newly discovered morbillivirus.

During a recent disease outbreak among harbour seals (Phoca vitulina) in the North and Baltic seas, more than 17,000 animals have died. The clinical symptoms and pathological findings were similar to those of distemper in dogs. Based on a seroepizootiological study, using a canine distemper virus (CDV) neutralization assay, it was shown that CDV or a closely related morbillivirus (phocid distemper virus-PDV) was the primary cause of the disease. The virus was isolated in cell culture from the organs of dead seals and characterized as a morbillivirus by serology (immunofluorescence neutralization and enzyme-linked immunosorbent assays) and by negative contrast electron microscopy. Experimental infection of SPF dogs resulted in the development of mild clinical signs of distemper and CDV-neutralizing antibodies. The disease was reproduced in seals by experimental inoculation of organ material from animals that had died during the outbreak. However, seals that had been vaccinated with experimental inactivated CDV vaccines were protected against this challenge. This fulfilled the last of Koch's postulates, confirming that the morbillivirus isolated from the seal organs, was the primary cause of the disease outbreak. The recent demonstration of the presence of a similar virus in Lake Baikal seals (Phoca sibirica), which infected these Siberian seals 1 year before the northwestern European seals were infected, raises new questions about the origin of this infectious disease in pinnipeds.

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Evaluation of enzyme-linked immunosorbent assays based on monoclonal antibodies for the serology and antigen detection in canine parvovirus infections.

An enzyme-linked immunosorbent assay (ELISA) system was developed for the detection of canine parvovirus (CPV) or CPV antigen in dog faeces and two other ELISA systems were developed for the detection of CPV-specific antibodies in dog sera. The ELISA's were based on the use of CPV-specific mouse monoclonal antibodies, which recognise different epitopes of the haemagglutinin of CPV and which also neutralise the virus. A double antibody sandwich (DAS) ELISA for the detection of CPV in dog faeces was compared with the haemagglutination (HA) test. The DAS-ELISA proved to be more specific, sensitive and easier to perform than the HA assay. An indirect ELISA and a competitive ELISA for the detection of CPV-specific antibodies in dog sera were compared with the haemagglutination inhibition (HI) test. Both ELISA systems proved to be specific and easy-to-use methods for the detection of CPV-specific antibodies. The indirect ELISA, specially, proved to be more sensitive than the HI test. The higher sensitivity and specificity of the ELISA's as compared to HA and HI tests, and their ease of use, make them suitable for routine use in the serology and diagnosis of CPV infections.

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Comparison of enzyme-linked immunosorbent and virus neutralization assays for the serology of reovirus infections in laboratory animals.

An enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of reovirus-specific IgM and IgG serum antibodies in rodents, detecting reovirus group reactive antibodies, was compared with reovirus types 1, 2 or 3 specific virus neutralization (VN) assays. To this end, serum samples were collected from specific pathogen-free (SPF) BALB/c RIVM mice, after experimental infection with any of the three mammalian reovirus serotypes. The majority (seven out of twelve) of the reovirus type 1-infected and one third (two out of six) of the reovirus type 3-infected mice died within 15 days after infection, whereas all (six out of six) of the reovirus type 2-infected animals survived. Using reovirus type 2 antigen in the ELISA, reovirus-specific IgM antibodies could be demonstrated within 1 week after infection in all the mice infected with reovirus types 2 or 3 and in the majority of the mice infected with type 1. Similarly, reovirus-specific IgG could be detected within 3 weeks in all the surviving mice infected with reovirus types 2 or 3 and within 5 weeks in all surviving mice infected with reovirus type 1. These results coincided well with the data obtained in the VN assays, in which all the infected animals also could be detected within 1 week after infection. As expected, titers were detected most rapidly and to the highest levels in the homologous VN assays. Given the sensitivity and the specificity of the ELISA system demonstrated in this paper and its suitability for incorporation in automated systems, the ELISA system should be considered valuable for the routine serologic diagnosis of reovirus infections in rodents.

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An enzyme linked immunosorbent assay (ELISA) for the detection of feline immunodeficiency virus (FIV) antigen in cell culture and FIV specific antibodies in feline serum.

Recently, a newly discovered lentivirus--Feline Immunodeficiency Virus (FIV)--was identified as the cause of an AIDS-like syndrome in the cat. For the detection of FIV antigen in cell culture, an antigen capture ELISA was developed. In this assay FIV specific IgG was used as a capture antibody and the same preparation coupled to biotine as a conjugate. With this test viral antigen could be demonstrated in FIV infected cell cultures within four days after infection and the assay proved to be at least as sensitive as a reverse transcriptase test in demonstrating virus infected cultures. A modification of this assay in which a titrated amount of FIV antigen was incubated with dilutions of serum samples of naturally or experimentally infected cats, allowed the detection of FIV specific antibodies. In a serological survey, conducted among cats with different clinical histories in The Netherlands and The Federal Republic of Germany, the antibody inhibition ELISA was compared with a commercially (IDEXX) available indirect ELISA system. It was shown that both assays identified nine seropositive cats among 200 animals with a history of undefined chronic disease, whereas no positive reactions were obtained with 65 serum samples from apparently healthy animals. Both assays also gave positive results with serum samples from experimentally infected cats, within five weeks after infection. Of an additional panel of eight serum samples which had been found positive in the IDEXX indirect ELISA system, only five were found positive in the antibody inhibition ELISA and western blotting assay.(ABSTRACT TRUNCATED AT 250 WORDS)

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[Another case of Hantaan virus infection in The Netherlands].

In a serological survey among Dutch patients suspected of leptospirosis, using a recently developed enzyme-linked immunosorbent assay, a patient was traced with a high antibody titre to Hantaan virus. No anti-leptospira antibodies were detected in this 27-year-old man. Shortly before he had been admitted to the hospital with progressive dyspnoea and coughing, accompanied with high fever. An interstitial pneumonia was diagnosed. He subsequently developed a progressive renal failure with proteinuria and polyuria. Later a liver failure accompanied with thrombocytopenia, anaemia and coagulation disturbances occurred. Before an aetiological diagnosis was made, the patient was treated with erythromycin. The patient eventually recovered completely. Based on the clinical symptoms and the positive serology, it was concluded that the disease diagnosed had probably been caused by a Hantaan virus infection. The diagnostic value of Hantaan virus serology in patients with similar symptoms is stressed.

Adult↗

Comparison of immunofluorescence and enzyme-linked immunosorbent assays for the serology of Hantaan virus infections.

Three enzyme-linked immunosorbent assay (ELISA) systems based upon different principles were developed for the serology of Hantaan virus infections and compared with an indirect immunofluorescence assay (IFA). The indirect IFA was carried out with gamma-irradiated Hantaan virus-infected and uninfected Vero E6 cells fixed with ethanol (-70 degrees C) or acetone (20 degrees C) on drop slides and a FITC-coupled sheep anti-human Ig preparation. Atypical staining in the IFA was avoided by using ethanol (-70 degrees C) instead of acetone (20 degrees C) fixation. In the first ELISA ('cell-assay'), Hantaan virus-infected or uninfected Vero E6 cells were used as antigens, which after gamma-irradiation were seeded into microtiter ELISA strips. Serial dilutions of human sera were incubated and specific antibodies were demonstrated with a horseradish peroxidase (HRPO)-conjugated sheep anti-human Ig preparation. In the second ELISA ('competition-assay') an affinity-purified human Ig preparation was used as a capture antibody for Hantaan virus antigen. After incubation of serial dilutions of human sera with this coat, the reactivity of the affinity purified anti-Hantaan virus Ig coupled to HRPO was determined. In the third ELISA ('complex trapping blocking [CTB]-assay') the same capture antibody was used to react with a mixture of the antigen and serial dilutions of human sera. The reactivity with the same HRPO conjugate was then determined. The results obtained in the respective assay systems with sera from people at risk or suspected of Hantaan virus infection coincided well. The CTB-ELISA proved to be faster and more sensitive than both the other ELISA systems, without giving more non-specific reactions: it detected almost all the IFA positive samples.(ABSTRACT TRUNCATED AT 250 WORDS)

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Vaccination of harbour seals (Phoca vitulina) against phocid distemper with two different inactivated canine distemper virus (CDV) vaccines.

Two inactivated canine distemper virus (CDV) vaccines--an adjuvanted whole inactivated virus and a subunit ISCOM preparation--were tested for their ability to induce protective immunity in harbour seals (Phoca vitulina) against phocid distemper, a disease that recently killed greater than 17,000 harbour seals in the North and Baltic seas, and was shown to be caused by infection with a newly discovered morbillivirus, which is antigenically closely related to CDV. Four CDV seronegative harbour seals were vaccinated three times with the whole-virus vaccine, two with the ISCOM subunit vaccine and two were sham-vaccinated with an antigen-free preparation. Ten days after the last vaccination, when all six vaccinated animals had developed CDV neutralizing antibody titres ranging from 300 to 3000, all eight animals were challenged by the oculonasal and the peritoneal routes, with an organ suspension from dead seals. None of the six vaccinated animals developed clinical signs. The two sham-vaccinated seals died on days 14 and 18, respectively, after having shown a body temperature rise, respiratory symptoms and weight loss. In organs from both dead animals morbillivirus antigen was demonstrated with an enzyme-linked immunosorbent assay and an immunofluorescence assay. One of these two animals had developed a low titre of CDV-specific antibodies just before death. These data clearly indicate that seals can be protected from fatal challenge with the phocid distemper virus (PDV), by vaccination with certain inactivated CDV vaccines. They also reconfirm that infection with PDV should be considered the primary cause of the recent epizootic in seals.

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Comparison of an enzyme-linked immunosorbent assay, an immunofluorescence assay and a hemagglutination inhibition assay for detection of antibodies to K-papovavirus in mice.

The sensitivity of a newly developed enzyme-linked immunosorbent assay (ELISA) for detection of antibody to K virus was compared with the sensitivities of an immunofluorescence assay (IFA) and a hemagglutination inhibition assay (HIA). Specific pathogen-free BALB/c RIVM mice, 5 weeks old, were inoculated intraperitoneally with a mouse organ suspension containing 10(4.5) TCID50 of K virus per dose. Control animals were inoculated with a control mouse organ suspension. No clinical signs were observed during the 7 weeks they were followed for the development of serum antibody. The ELISA proved to be the most sensitive of the three assays and demonstrated K virus-specific antibodies as early as 3 days after infection.

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[An outbreak of dog distemper among seals (2)].

Serological findings which had showed that the primary cause of the recent outbreak of serious disease of seals in the seas of North-Western Europe, is infection with canine distemper virus, were confirmed by in vivo and in vitro isolation of the virus from seals which had died at different locations during the outbreak. The virus which proved to be pathogenic for dogs, was characterised as canine distemper virus on the basis of immunofluorescence, virus neutralisation and electron microscopical studies.

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Purification of infectious canine parvovirus from cell culture by affinity chromatography with monoclonal antibodies.

Immuno affinity chromatography with virus neutralizing monoclonal antibodies, directed to the haemagglutinating protein of canine parvovirus (CPV) was used to purify and concentrate CPV from infected cell culture. The procedure was monitored by testing the respective fractions in an infectivity titration system, in an ELISA, in a haemagglutination assay and by negative contrast electron microscopy to quantify CPV or CPV antigen. The degree of purification was further estimated by testing the fractions for total protein content in a colorimetric method, for bovine serum albumin content in an ELISA and by SDS-PAGE. Over 99% of the contaminating proteins proved to be removed, and 20% or 70-90% of infectious CPV or CPV antigen, respectively, was recovered.

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Selective inhibitory effects of (S)-9-(3-hydroxy-2-phosphonyl-methoxypropyl) adenine and 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodouracil on seal herpesvirus (phocid herpesvirus 1) infection in vitro.

From a selection of 25 antiviral compounds with specific anti-herpes activity or broad-spectrum antiviral properties, two compounds, namely (S)-9-(3-hydroxy-2-phosphonyl-methoxypropyl)adenine and 1-(2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl)-5-iodouracil, appeared particularly effective in inhibiting the cytopathogenicity of seal herpesvirus (phocid herpesvirus 1).

Adenine↗

The isolation and partial characterization of a highly pathogenic herpesvirus from the harbor seal (Phoca vitulina).

This report describes the first isolation and partial characterization of a herpesvirus from the harbor seal (Phoca vitulina). The virus was isolated during a disease outbreak in a group of young seals nursed in a seal orphanage in The Netherlands. Almost half of the seals died with symptoms of acute pneumonia and focal hepatitis and the virus was isolated of organs of most of the dead animals. Seven out of ten seals of which paired serum samples were obtained showed seroconversion in a virus neutralization test during this outbreak. The virus was tentatively characterized as a herpesvirus (seal herpesvirus: SeHV or phocid herpesvirus 1) on the basis of its characteristic morphology in electron microscopy, buoyant density in sucrose, sensitivity to ether and heat treatment and its antigenic relationship with other probable members of the Alphaherpesvirinae subfamily. The virus caused cytopathic changes within 24 hours after inoculation in seal kidney cells, consisting of a focal rounding of cells and syncytium formation. No cytopathic changes were observed in the cells of nine other mammalian species tested.

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