- Volume 55, Issue 1, 1981
Volume 55, Issue 1, 1981
- Animal
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Rapid Detection of Human Viruses in Faeces by a Simple and Routine Immune Electron Microscopy Technique
More LessSummaryAn immune electron microscopy technique modified from the Anderson & Doane (1973) method by using commercial pools of human gamma-globulins has been successfully used for the morphological identification of several different viruses from clarified faeces. This technique is simple, rapid and numerous samples can be easily processed.
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Biological Properties and Carbohydrate Composition of Human Parainfluenza Virus Type 1 in Two Host Systems
More LessSummaryHuman parainfluenza virus type 1 (HA2 virus) grown either in embryonated hens’ eggs or in M. rhesus monkey kidney cells showed differences in size, biological properties and carbohydrate composition. Egg-grown virus showed a larger size (233 nm versus 167 nm), a higher neuraminidase activity (specific activity and initial and maximum velocity) and a higher haemolytic activity than monkey kidney cell-grown virus. The haemagglutinin titre was identical for the HA2 strain grown in both host systems when tested with human O Rh+, guinea-pig and hen red blood cells, but reduced by more than 100-fold when tested with grivet monkey red blood cells. In addition, the carbohydrate content (mainly neutral sugars) was higher in egg-grown virus (9.2%) than in virus grown in MK cells (5.7%), and the amino to neutral sugar ratio was lower (1.2 versus 2.1). The sugars were identified as fucose, mannose, galactose, glucose, glucosamine and galactosamine. The prominent neutral monosaccharide was glucose in egg-grown virus and fucose in MK cell-grown virus. HA2 virus infection of MK cells increased fucose and glucose, and decreased mannose and galactose levels.
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Molecular Cloning of Infectious DNA from Human Papovavirus BK in Escherichia coli
More LessSummaryRecombinant DNA constructed from unit length BK virus DNA and from several defective viral forms was cloned in Escherichia coli HB101. The cloned unit length BK virus DNA retained its infectivity for human embryonic kidney cells, whereas the cloned defective DNA showed no infectivity. Restriction endonuclease digestion of cloned defective DNA and purified virus DNA indicated that many of the defective forms contain re-iterated sequences.
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- Plant
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Indirect ELISA Methods for the Broad Specificity Detection of Plant Viruses
More LessSummaryIndirect ELISA on plates not precoated with antibodies enabled the detection of cross-reactions among a wider range of serologically related viruses in the tymo-, tombus-, como-, tobamo-, potex-, carla- and potyvirus groups than was possible with the direct double-antibody-sandwich method. Used with purified virus preparations and unfractionated antisera which have to be preabsorbed with crude plant sap the method seems promising for the detection of serological relationships among plant viruses, especially when the latter can be purified only with a low yield or when antisera are in short supply and immuno-electron microscopy is not possible. The quantitative outcome of the test was influenced by the concentration of the reactants, the purity and specific adsorption characteristics of the virus and the length of the immunization period. Strains of Andean potato latent virus (APLV) were detected reliably in crude plant sap when indirect ELISA was done on plates precoated with antiviral antibodies from rabbit or guinea-pig (‘first antibodies’). The trapped virus particles were reacted with antiviral antibodies from chicken (‘second antibodies’) which were then detected by enzyme-labelled rabbit anti-chicken antibodies. Other combinations of ‘first’ and ‘second’ antibodies resulted in non-specific reactions in the presence of crude plant sap. The specificity of direct ELISA was not sufficiently broadened to enable the routine detection of APLV strains when antisera were used which, due to altered immunization schemes, were expected to have an especially broad cross-reactivity.
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Cynosurus and Cocksfoot Mottle Viruses: a Comparison
More LessSummaryThe biological, physical and chemical properties of New Zealand isolates of cocksfoot mottle (CfMV) and cynosurus mottle (CyMV) viruses were compared. The two viruses could be distinguished by host range differences and serological properties but their physical and chemical properties were similar. The virus particles differed in their buoyant density in CsCl (1.386 g/ml for CfMV; 1.379 g/ml for CyMV) and in their banding behaviour in Cs2SO4 gradients (CfMV formed two bands at pH 5 while CyMV formed only one). Furthermore, CyMV was degraded by low concentrations of SDS at pH 7, while CfMV was resistant except in the presence of EDTA. The viral RNAs were comparable in mol. wt. but differed in their amount of secondary structure and sedimentation values. Molecular hybridization studies using 3H-labelled complementary DNA (cDNA) probes indicated that the RNA genomes of the two viruses had between about 5% and 8% of their base sequences in common. Both viruses were detected in the nuclei of infected cells. However, CfMV, unlike CyMV, formed crystalline arrays in the cytoplasm. It is concluded that although New Zealand isolates of CyMV and CfMV share a number of similar properties they are distinct viruses.
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Two Serologically Unrelated Viruses Isolated from a Phialophora sp.
More LessSummaryTwo isometric viruses, with similar diameters (34 to 36 nm), obtained from an isolate of a Phialophora sp. with lobed hyphopodia were separated by salt gradient elution from DEAE-cellulose and from SP-Sephadex. Virus A had a sedimentation coefficient of 116S, three double-stranded RNA components of mol. wt. 1.29 × 106, 1.22 × 106 and 1.03 × 106 and one major capsid polypeptide, mol. wt. 60000. Virus B had a sedimentation coefficient of 122S, three double-stranded RNA components of mol. wt. 1.32 × 106, 1.25 × 106 and 1.03 × 106 and one major capsid polypeptide, mol. wt. 66000. The two viruses were unrelated serologically to each other and to 13 viruses obtained from a related fungus Gaeumannomyces graminis var. tritici.
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Infection of Cowpea Protoplasts with Both the Common Strain and the Cowpea Strain of TMV
More LessSummaryThe common strain of tobacco mosaic virus (TMV) which does not multiply in cowpea plants, can infect isolated cowpea mesophyll protoplasts to a level of 50 to 70%. The highest percentage of infection was obtained when both the virus and the protoplasts were preincubated with poly-d-lysine and inoculation was with 1 µg/ml virus, 1 µg/ml poly-d-lysine, 2 × 105 protoplasts/ml in 0.6 m-mannitol, 0.1 m-potassium phosphate pH 5.4 at 0 °C for 7.5 min. The rate of virus multiplication in the infected protoplasts proceeded rather slowly and the increase in the percentage of infected protoplasts extended over 40 h. This can be explained by assuming a variable dose of infecting virus particles resulting in a considerable asynchrony in the infection process for different protoplasts. The course of infection of the common strain of TMV in cowpea protoplasts is similar to that of the cowpea strain of TMV, which has cowpea as a natural host.
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