SUMMARY A simple and rapid procedure for identifying adenovirus and rotavirus in stool extracts has been developed. The technique is based on polyacrylamide gel electrophoresis of the virus nucleic acid, but sample preparation is straightforward and does not entail phenol extraction or the use of a radioactive label. Furthermore, processing is not influenced by the amount of specimen obtained and is thus suitable for application as a batch testing method. This approach removes the need for specific antisera, which are not readily available since most of these viruses cannot be grown using routine tissue culture procedures. Trials in this laboratory have indicated that the technique is of comparable sensitivity to electron microscopy.

Newcastle upon Tyne, and the tDepartment ofBiology, Sunderland Polytechnic, Sunderland, Tyne and Wear SUMMARY A commercially available latex agglutination test, Rotalex (Orion Diagnostics, Finland), for detecting rotaviruses was evaluated in comparison with four other tests (electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay) routinely used in our laboratories. Although Rotalex was the least complex method, it showed lack of specificity and sensitivity when carried out according to the manufacturer's instructions. Four basic modifications of Rotalex are described. These include the use of Hank's balanced salt solution, increasing the incubation time to 20 min, reading the agglutination result by an experienced observer, and the use of 50 mm square glass plates. The modified procedure gave results which were comparable with those obtained by electron microscopy, immunofluorescence, polyacrylamide gel electrophoresis, and enzyme linked immunosorbent assay. The latter techniques, when used to detect rotavirus, all gave similar results.

SUMMARYThis report describes the development of a solid-phase haemadsorption system using chromic chloride-linked, antibody coated erythrocytes. It is proposed to call this technique solid phase aggregation of coupled erythrocytes (SPACE). The system is suitable for the detection of virus antigens, such as from rotavirus infections, which are present in 'dirty' or 'mixed' preparations such as faeces, urine or exudates. The test uses microtitre U-form plates coated" with specific antivirus antibody; faecal suspensions are added and virus or antigen allowed to adsorb. The plates are then washed and adsorbed antigens are detected by the addition of virus-specific IgG-coated erythrocytes. The resultant settling pattern is read in the same manner as a conventional haemagglutination test. The system is compared with electron microscopy and fluorescent antibody techniques.

If trypsin is incorporated in the tissue culture medium it is possible to carry out a sensitive immunofluorescence assay for the presence of human rotavirus. The enhanced effect of trypsin is negated by serum. It has also been established that naturally occurring enzymes in faeces enable some virus to penetrate tissue culture cells. The role of these naturally occurring enzymes in the pathogenesis of rotavirus infection is discussed.