Dried Blood Spots as a Clinical Samples for Laboratory Diagnosis and Surveillance of Vaccine-Preventable Diseases in Bulgaria

Stefka Krumova, Elitsa Golkocheva-Markova, Svetla Angelova, Silviya Voleva, Adelina Pavlova, Irina Georgieva, Petia Genova-Kalou


In recent years the dried blood spots (DBS) had new and innovative applications in medicine, neonatology, virology and microbiology. This study aimed to evaluation of the frequency of detection of viral IgM/IgG markers in dried blood spots and introducing an easy-to-implement protocol for serum extraction in measles, mumps and rubella surveillance. The total 204 clinical samples (102 serum samples and 102 dried blood spots) collected from 102 patients were included. All specimens were tested for presence of specific viral markers (IgM and IgG antibodies) by a commercial indirect enzyme-linked immunosorbent assay (ELISA). Of all tested patients, three (3/102, 2.94%, 95% CI: 0 ÷ 6.22) were confirmed for acute measles infection and two (2/102, 1.96%, 95% CI: 0 ÷ 4.65) for mumps. Double positive ELISA-IgM results were found in their serum samples and DBS. No acute rubella infection and rubella IgM marker were detected in both clinical samples. By immunoassay analysis of all 102 patients, measles, mumps and rubella IgG were found in 83/102 (81%, 95% CI: 73.40 ÷ 88.60), 76/102 (75%, 95% CI: 66.60 ÷ 83.40) and 79/102 (77%, 95% CI: 68.83 ÷ 85.17) serum samples.  Comparative results were obtained in the adequately obtained DBS. Viral IgG seroprevalence in DBS were obtained in 79/102 (77%, 95% CI: 68.83 ÷ 85.17) for measles, 69/102 (68%, 95% CI: 58.67 ÷ 77.33) for mumps and 73/102 (72%, 95% CI: 63 ÷ 81) for rubella, respectively. Double negative results for each screened viral markers were proven in six tested patients.

The study shown higher extinction value (Ratio and NovaTec units) in DBS compared to serum samples of same persons were calculated. Our studies show over 90% coincidence in combined ELISA assay of viral markers against measles, mumps, and rubella in serum samples and DBS. DBS clinical approach is non-aggressive and more acceptable to the public (including young children, pregnant women, etc.). It has a variety of new and innovative applications in medicine and in particular in the laboratory diagnosis of acute and past (presence of protective immunity) measles, mumps and rubella infection in the phase of elimination.


dried blood spots; vaccine-preventable diseases; measles; mumps; rubella; ELISA IgM/IgG.

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