Rapid detection of Australia/SH antigen and antibody by a simple and sensitive technique of immunoelectronmicroscopy

Abstract

A simple and practical method is presented for demonstrating the presence of the Australia/SH antigen and its corresponding antibody in serum specimens, both qualitatively and quantitatively. The method is based on the electronmicroscopic visualization of characteristic aggregates of antigen–antibody complexes formed in the mixture of a serum specimen and the appropriate Australia/SH detector reagent. It involves the use of a microtechnique requiring minute amounts of reagents and provides, as a result of diffusion and filtration through agar gel, partially purified and concentrated preparations, ready for electronmicroscopic examination in less than an hour. The method is highly specific and yields reproducible results. Its sensitivity was found to be greater than that of the crossover electrophoresis test and closely approximates that of the complement fixation test, with the added advantage of not being affected by the "prozone phenomenon." The method can be recommended for use in laboratories equipped with electronmicroscopic facilities to establish a differential diagnosis of viral hepatitis cases, perform rapid screening of blood samples (blood products) for the presence of Australia/SH antigen, and clarify equivocal results obtained by other methods. It is expected that the agar–diffusion–filtration technique will also prove useful, in general, for enhancing the chances of detecting virus particles in suspensions of relatively low virus concentrations.

Antipili antibody affords protection against experimental ascending pyelonephritis.

“…Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was done by the method of Laemmli (9). Isolated pili were negatively stained with 0.5% uranyl acetate by an agar diffusion method (10). Agar gel double diffusion was performed according to the method of Ouchterlony and Nilsson (11).…”

Heat-Resistant Infectivity of Herpes Simplex Virus Revealed by Viral Transfection

“…The con centration-dependent effect of DMSO in en hancing the infectivity of HSV DNA [16] was also obvious when heated virus was trans fected.…”

“…Samples were processed for negative staining with 3% phosphotungstic acid [16], Processing and exami nation of samples were kindly carried out by Dr. Cor nelia Gatihl, Institute of Hygiene. University of Frank furt Main.…”

Rapid Detection of Human Viruses in Faeces by a Simple and Routine Immune Electron Microscopy Technique

“…Various IEM approaches involving the centrifugation of immune complexes (Almeida & Waterson, 1969), the use of agar (Kelen et al, 1971;Anderson & Doane, 1973) or antibody-coated grids (Derrick, 1972;Shukla & Gough, 1979) have been described. We report here a simple and rapid IEM technique modified from the Anderson & Doane (1973) method of serum-in-agar diffusion where specific antisera were substituted by commercially available pools of human gamma-globulins.…”