http://repub.eur.nl/res/aut/8641/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/35326/ 2007-07-01T00:00:00Z Both rhesus and cynomolgus macaques have been used as animal models for measles I vaccination and immunopathogenesis studies. A number of studies have suggested that experimental measles virus (MV) infection induces more-characteristic clinical features in rhesus than in cynomolgus monkeys. In the present study, both macaque species were infected with two different wild-type MV strains and clinical, virological and immunological parameters were compared. The viruses used were a genotype C2 virus isolated in The Netherlands in 1991 (MV-Bil) and a genotype B3 virus isolated from a severe measles case in Sudan in 1997 (MV-Sudan). Following infection, all rhesus monkeys developed a skin rash and conjunctivitis, which were less obvious in cynomolgus monkeys. Fever was either mild or absent in both species. Virus reisolation profiles from peripheral blood mononuclear cells and broncho-alveolar lavage cells and the kinetics of MV-specific IgM and IgG responses were largely identical in the two animal species. However, in animals infected with MV-Sudan, viraemia appeared earlier and lasted longer than in animals infected with MV-Bil. This was also reflected by the earlier appearance of MV-specific serum IgM antibodies after infection with MV-Sudan. Collectively, these data show that cynomolgus and rhesus macaques are equally susceptible to wild-type MV infection, although infection in the skin seems to follow a different course in rhesus macaques. MV-Sudan proved more pathogenic for non-human primates than MV-Bil, which may render it more suitable for use in future pathogenesis studies. http://repub.eur.nl/res/pub/39691/ 2005-04-01T00:00:00Z Real-time detection of polymerase chain reactions allows convenient detection and quantification of virus-derived nucleic acids in clinical specimens. We have developed a real-time RT-PCR assay for the detection of measles virus (MV) genomic RNA, and compared it to a well-established conventional RT-PCR assay. Based on a serial dilution of the live-attenuated MV Edmonston Zagreb vaccine, the detection limits were approximately 0.1 and 0.02 cell culture infectious dose 50% units (CCID50) per test for the conventional and TaqMan RT-PCR assays, respectively. Furthermore, tissue materials spiked with known quantities of MV were equally well detected in both assays. The TaqMan assay was linear within a range of 104.4to 10-0.6CCID50/ml, with an intra-assay variability lower than 3% and an inter-assay variability ranging from 1.5% at 104.4CCID50/ml to 8.7% at 10-0.6CCID50/ml. The TaqMan assay could detect representative wild-type viruses from the currently active MV clades, and could detect MV genome in clinical specimens obtained from measles patients. Finally, quantification of MV RNA in peripheral blood mononuclear cells or broncho-alveolar lavage cells from cynomolgus macaques collected at different time points after experimental infection showed a good correlation with virus isolation data. In conclusion, the TaqMan assay developed is specific, sensitive, rapid and reproducible, and can be of use for diagnostic purposes or for studies on the pathogenesis of measles. http://repub.eur.nl/res/pub/3950/ 2004-08-01T00:00:00Z Dried blood spots collected on filter paper are considered potential clinical specimens for measles surveillance because of their ease of collection, storage, and transport. The usefulness of these samples for surveillance of measles was evaluated in a field setting. Blood spots were collected by finger-prick from 316 clinically diagnosed measles patients in suburban Khartoum, mostly within a week after onset of the rash. Samples were collected between October, 2000 and April, 2003, and stored at 4 degrees C. Measles virus-specific IgM antibodies were detected in 200 (63%) of the samples using an "in-house" IgM capture ELISA. For 201 samples reconstitution and IgM measurement was repeated 1 year after initial testing with essentially the same results, showing the stability of IgM in the filter paper under these conditions. In a limited number of samples (n = 38) measles virus-specific IgM was also tested with a commercial indirect IgM ELISA. Although the results of the two assays correlated well, the "in-house" IgM capture ELISA proved slightly more sensitive. Measles virus-specific reverse transcriptase polymerase chain reaction (RT-PCR) amplicons were obtained from 16 of 57 (28%) samples tested. Sequencing of the 3' 456 nucleotides of the nucleoprotein gene showed the continued endemic circulation of genotype B3 viruses identified previously in this region. Although problems related to limited sample quantities were encountered, the present study confirms the usefulness of dried blood spots for measles surveillance. The results also demonstrate that measles continues to be endemic in the Sudan. http://repub.eur.nl/res/pub/39723/ 2001-03-21T00:00:00Z Despite the availability of safe and effective live attenuated vaccines, measles continues to be endemic in many developing countries. Control and elimination of measles will be especially difficult in East Africa, because of its limited infrastructure and political instability. We have studied diagnostic and epidemiological aspects of measles in suburban Khartoum, Sudan. Prospective studies were carried out in a cohort of clinically diagnosed measles cases and in a cohort of new-borns, which were both followed up for 2 years. The studies intended to provide a rational basis for improvement of measles vaccination strategies, and strengthen measles research infrastructure in Khartoum.