Serum samples were collected from 27 individuals who had been infected with a member of the genus Hantavirus in the Netherlands or Belgium during the last 15 years. These samples were tested in an immunofluorescence assay (IFA) and two enzyme-linked immunosorbent assay (ELISA) systems, using different virus strains that represented each of the four recently proposed serotypes of this genus. The serum samples from 11 individuals who had been infected through contacts with laboratory rats showed the highest reactivities with Hantaan virus (serotype I) and SR-11 (serotype II) in the IFA and ELISA systems. The samples of 16 individuals who had probably been infected through contacts with wild rodents showed the highest reactivities with Hällnäs virus (serotype III) in the IFA. All except two of these also showed the highest reactivity with Hällnäs virus in the two different ELISA systems.

Potency control of inactivated rabies vaccines for human and veterinary application is usually undertaken by vaccination-challenge tests (e.g. the mouse potency test). For practical and ethical reasons there is an urgent need to replace in vivo potency control procedures, at least in part, by reliable methods of in vitro potency testing. Quantitative ELISA systems for potency control were developed using monoclonal antibodies (MAbs) directed to the glycoprotein (GP) and to the nucleoprotein (NP) of the virus. Although immuno-capture and competition ELISAs for GP measurement had almost equal sensitivity (detection level GP < 0.1 IU), the competition data showed the best correlation with potency values when a panel of rabies vaccines for human use was tested (r = 0.88, n = 10). The NP values for this panel of vaccines in the competition system (detection level NP < 1 IU) also correlated well with potency values (r = 0.90, n = 10). The competition system proved to be best also with liquid adjuvanted veterinary rabies vaccines of LEP, PM, PV and SAD origin. The implementation of ELISA systems for potency control of rabies vaccines is discussed.

Since rabies is still a major cause of human death in many developing countries and the implementation of recommended post-exposure prophylaxis by vaccination and specific immunoglobulin therapy is largely hampered by its high cost, the development of cheap rabies vaccines and immunoglobulin preparation are a high priority in these countries. In this paper various purification methods of equine rabies immunoglobulin based on different principles are compared with respect to their effect on final yield and biological activity. It is shown that a combination of ammonium sulphate (AS) precipitation and DEAE ion exchange chromatography results in an acceptable recovery rate of biological activity and a product of relatively high purity. Although affinity chromatography with protein G in combination with AS precipitation results in a similar recovery rate and a product of considerably higher purity, the cost of this procedure may be prohibitive for routine use in most developing countries. The effects of pepsin digestion time on the biological activity of the product and on the reduction of intact horse Ig are also studied. The desirability of this digestion procedure with respect to reduction of adverse side effects and efficacy of the product for post-exposure treatment is discussed.

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