Dried blood spot cards: A reliable sampling method to detect human antibodies against rabies virus

Background Although preventable by vaccination for more than a century, rabies virus still causes numerous fatalities every year. To determine antibody levels in humans, blood collected with a finger prick and applied on dried blood spot (DBS) cards is an alternative for venipuncture. The use of DBS is specifically valuable in remote areas, as it is easy to perform, store and transport. Therefore, the technique is frequently used for epidemiological studies of tropical diseases. Up to present, determination of rabies virus antibody levels on human DBS has not been validated. Methodology/Principal findings We evaluated the use of human DBS for rabies serology and analyzed 99 pre- or post-vaccination serum and DBS samples with a fluorescent antibody virus neutralization test (FAVNt), which is the gold standard to detect protective antibody levels, and a Bio-Rad Platelia Rabies II ELISA. Sensitivity and specificity of DBS eluates tested with the FAVNt were 97% and 92%, respectively and 87% and 96% when tested with the Platelia-II ELISA. Antibody levels measured in serum with the FAVNt, correlated best with antibody levels measured in DBS with the FAVNt (R = 0.88). Conclusions/Significance This is the first study that applies DBS for reliable detection of human antibodies against rabies virus. Both the FAVNt and Platelia-II ELISA demonstrate an acceptable performance on DBS, providing opportunities for rabies serology in remote areas. This technique could drastically ease studies evaluating (novel) rabies vaccination strategies and monitoring persisting immunity in humans at risk, living in rabies endemic regions. Author summary Rabies is a nearly 100% fatal disease in humans. However, available vaccines are effective in preventing rabies infection. To investigate if a person is protected against rabies, rabies virus neutralizing antibody levels in the blood are determined. The World Health Organization defines protective immunity as a rabies virus antibody concentration of at least 0.5 IU/ml detected in serum using a virus neutralization test. Yet, in remote areas serum may be rather difficult to collect, process and transport. Whole blood collected with a finger prick and applied on filter paper cards, also known as dried blood spots (DBS), are an easier alternative. This collection method is frequently used for serology of several tropical infectious diseases, but never studied for rabies serology in humans. Therefore, we compared antibody levels measured in serum with those measured in DBS eluates, using the gold standard FAVNt and related it to another commonly used test for human rabies serology, the Platelia-II ELISA. We found that both assays had a good performance on DBS eluates. The reported high specificities provide confidence that unprotected individuals will rarely be missed. Therefore, the DBS is a promising sampling technique for evaluations of vaccination strategies and monitoring persisting immunity after vaccination in populations at risk for rabies.