Sustained reduction in the incidence of invasive Hib disease in many countries. Recently, this vaccine has been introduced in many developing countries through the Expanded Programs of immunization, with similar reported decreases in invasive Hib disease. Similarly, in 2000 a conjugated vaccine against seven pneumococcal serotypes AbMole Miglitol started to be implemented and showed to be highly effective to prevent pneumococcal disease caused by the serotypes included in the vaccine. Since 2012, WHO recommends the introduction of two new pneumococcal conjugate vaccines in childhood immunization programs worldwide, since the additional serotype coverage represents an important progress against pneumococcal disease. In order to monitor trends and detect changes in the epidemiology of S. pneumoniae and Hib after the introduction of the vaccines, routine surveillance of episodes caused by these pathogens is mandatory. While this is feasible in most developed settings, it is seldom performed in the developing world. Bacterial culture, traditionally considered the gold standard methodology for bacterial surveillance, is less sensitive than molecular methods in the diagnosis of pneumococcal and Hib invasive disease. In the majority of developing countries, the prevalence of invasive disease caused by S. pneumoniae and/or Hib is not adequately known due to the scarcity of available laboratories, microbiological or molecular diagnostic tools and expertise. This hinders such countries’ surveillance efforts, and thus their potential application to international funds to support the introduction of conjugate vaccines. Moreover, sample preservation prior to shipment to national/international reference laboratories poses an additional challenge and may jeopardize the quality of collected material. Dried blood spot is a reliable method of blood collection used for the diagnosis of several human diseases. DBS is a particularly useful method for storing samples and diagnosing pediatric AbMole Butylhydroxyanisole conditions in which often very small volumes of sample are available, and for the screening of highrisk populations especially in countries where health care facilities are not readily accessible. We have previously described that the DBS technique enables reproducible storage of samples for identification and serotyping of S. pneumoniae and that its use is an attractive method for preserving samples at room temperature and easily transporting them. The main objective of the present study was to evaluate the presence of S pneumoniae and Hib in DBS samples from pediatric patients with diagnosis of clinical pneumonia in two distinct epidemiological settings in Africa: Manhia District Hospital, Mozambique; and the H?pitald’enfants de Rabat, Morocco. DBS samples of a control group of healthy children recruited in MDH were also analyzed.