Immune Electron Microscopy (IEM) of a Canine Adeno-Associated Virus

Author(s):  
Gunter F. Thomas ◽  
M. David Hoggan

In 1968, Sugimura and Yanagawa described a small 25 nm virus like particle in association with the Matsuda strain of infectious canine hepatitis virus (ICHV). Domoto and Yanagawa showed that this particle was dependent on ICHV for its replication in primary dog kidney cell cultures (PDK) and was resistant to heating at 70°C for 10 min, and concluded that it was a canine adeno-associated virus (CAAV). Later studies by Onuma and Yanagawa compared CAAV with the known human serotypes (AAV 1, 2, 3) and AAV-4, known to be associated with African Green Monkeys. Using the complement fixation (CF) test, they found that CAAV was serologically related to AAV-3 and had wide distribution in the dog population of Japan.

Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.


Intervirology ◽  
1974 ◽  
Vol 4 (2) ◽  
pp. 110-118 ◽  
Author(s):  
Stephen A. Locarnini ◽  
Allan A. Ferris ◽  
Anthony C. Stott ◽  
Ian D Gust

1983 ◽  
Vol 29 (1) ◽  
pp. 111-119 ◽  
Author(s):  
Pierre Payment ◽  
Reda Ayache ◽  
Michel Trudel

In this second study (1979–1981) of the viral content of sewage we have demonstrated the presence of poliovirus types 1, 2, and 3 in Laval and Montreal. Several strains of poliovirus types 2 and 3 were nonvaccinal. This is in contrast with our first study (1977–1978) in which only type 1 poliovirus isolates were nonvaccinal. Coxsackievirus types B-3, B-4, and B-5 and echovirus types 1, 7, and 11 were also isolated from sewage. Interestingly, these isolations coincided with reports of isolation of the same strains during the same period by diagnostic laboratories. Our method based on Vero and BSC-1 cell cultures for virus isolation and immune electron microscopy for identification permitted the recovery not only of several strains of enteroviruses but also of some adenoviruses and reoviruses.


Author(s):  
Linda M. Stannard ◽  
Margaret Lennon

Burnupena cincta and Fusus verruculatus are two whelks which inhabit the intertidal zones of the Cape Peninsula shore. Their respiratory pigments, or haemocyanins, are morphologically similar in structure (Figs. 1 and 2) and appear in the electron microscope as short cylindrical rods about 34 nm in diameter and 36 nm high. Viewed side-on the molecules show regular banding suggesting a structure composed of six equidistant rings of sub-units. Occasionally the particles have the appearance of possessing a central “belt” in the position of the 3rd and 4th rows of sub-units. End-on views of the haemocyanin molecules show a circular contour with a dense outer ring and a less dense inner ring in which 10 definite sub-units may frequently be distinguished. A number of molecules display an extra central inner component which appears either as a diffuse plug or as a discrete ring-shaped core ± 8 nm in diameter.


Author(s):  
J. P. Brunschwig ◽  
R. M. McCombs ◽  
R. Mirkovic ◽  
M. Benyesh-Melnick

A new virus, established as a member of the herpesvirus group by electron microscopy, was isolated from spontaneously degenerating cell cultures derived from the kidneys and lungs of two normal tree shrews. The virus was found to replicate best in cells derived from the homologous species. The cells used were a tree shrew cell line, T-23, which was derived from a spontaneous soft tissue sarcoma. The virus did not multiply or did so poorly for a limited number of passages in human, monkey, rodent, rabbit or chick embryo cells. In the T-23 cells, the virus behaved as members of the subgroup B of herpesvirus, in that the virus remained primarily cell associated.


Author(s):  
R. I. Johnsson-Hegyeli ◽  
A. F. Hegyeli ◽  
D. K. Landstrom ◽  
W. C. Lane

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).


Author(s):  
Charles D. Humphrey ◽  
E. H. Cook ◽  
Karen A. McCaustland ◽  
Daniel W. Bradley

Enterically transmitted non-A, non-B hepatitis (ET-NANBH) is a type of hepatitis which is increasingly becoming a significant world health concern. As with hepatitis A virus (HAV), spread is by the fecal-oral mode of transmission. Until recently, the etiologic agent had not been isolated and identified. We have succeeded in the isolation and preliminary characterization of this virus and demonstrating that this agent can cause hepatic disease and seroconversion in experimental primates. Our characterization of this virus was facilitated by immune (IEM) and solid phase immune electron microscopic (SPIEM) methodologies.Many immune electron microscopy methodologies have been used for morphological identification and characterization of viruses. We have previously reported a highly effective solid phase immune electron microscopy procedure which facilitated identification of hepatitis A virus (HAV) in crude cell culture extracts. More recently we have reported utilization of the method for identification of an etiologic agent responsible for (ET-NANBH).


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