Development of a novel diagnostic kit candidate to detect dengue antibody, using IgY and protein a positive Staphylococcus aureus

Abstract

Early diagnosis of dengue hemorrhagic fever (DHF) is very important in determining further management. Diagnostic kits are intended as relatively faster, easier, and cheaper diagnostic tools. The objectives of this experimental laboratory study was to prepare a diagnostic kit candidate to detect dengue antibody in human serum using co -agglutination methods by utilizing protein A positive Staphylococcus aureus as a carrier to bind Fc immunoglobulin fractions without altering its ability to bind antigen. To enhance the sensitivity and specificity of the diagnostic kit, chicken IgY was used as one of the components. Rabbit immunoglobulin chicken anti-IgY was used as an intermediary connection between protein A positive and Staphylococcus aureus with chicken IgY. Solution A contains Staphylococcus aureus Cowan I and rabbit immunoglobulin chicken anti-IgY serum while Solution B contains anti-dengue chicken serum and dengue antigen. The laboratory experiments produced a formula of A: B = 1~3: 1~3 (v/v), which was then tested to human serum and compared with IgM and IgG ELISA tests as the gold standard. Data on the results of the test were statistically analyzed using chi - square test. Of the 65 qualified samples used, 47.69% resulted in true positive and 41.54% resulted in true negative. Meanwhile, the remaining samples demonstrated false negative (6.15%), positive predictive value (91.2%) and negative predictive value (87.1%). The sensitivity and specificity were 89% and 90%, respectively, with no significant diffe rences (p>0.05). Therefore, co-agglutination method using S. aureus with protein A positive as a carrier can be proposed as a diagnostic kit candidate to detect human serum dengue antibody to screen dengue hemorrhagic fever cases.