Optimization of preservation conditions for the detection of adenosine deaminase isolated from salivary glands of field-collected aedes albopictus / Nurul Syahida Syamira Rusdi

Dengue fever, caused by dengue virus (DENV) is a public health concern since it was first recognized in the 1780s. Adenosine deaminase (ADA) presents abundantly in the salivary glands of Aedes albopictus, the secondary vector of DENV. It is a protein essential for viral transmission into a human host following the blood-feeding process. ADA provokes the host antigenic and immunogenic responses thus favoring the transmission of DENV upon pathogenesis. Improper sample collection and handling might cause sample degradation leading to false detection of relevant proteins. Hence, this study aims to determine the optimum preservation media and storage conditions for the detection of salivary ADA extracted from Ae. albopictus. Samples were preserved in a number of media (i.e. phosphate buffered saline, PBS; phosphate buffered saline supplemented with protease inhibitor, PBS-Pi; cell lysis buffer, CLB) at various temperature (i.e. -20°C, 4°C and room temperature) prior to protein profile analysis by SDS-PAGE. Protein profiles revealed that all three media preserved at -20°C and 4°C can be used for salivary glands preservation as the putative protein of interest was detected at molecular weight of 53 kDa. Further experiments using affinity chromatography or protein sequencing shoul be done to confirm the identity of the putative ADA protein.