The test is only valid if the migration control collection is visible. bacterially indicated recombinant protein rSP03B as antigen. For test optimization, pre-immune sera from non-bitten laboratory-bred Beagles were used as negative settings. In order to validate the test, sera from laboratory-bred Beagles experimentally exposed to bites were used as positive settings. Additionally, all samples were tested by ELISA using whole Rbin-1 salivary gland homogenate (SGH) and the rSP03B protein as antigen. An almost perfect degree of agreement was found between the ICT and the SGH-ELISA. Furthermore, the newly proposed rSP03B sero-strip showed a level of sensitivity Rbin-1 of 100% and Rbin-1 a specificity of 86.79%. Conclusions/Significance We developed a simple and quick ICT based on the rSP03B salivary protein, able to replace the standard ELISA used in earlier studies. Our rSP03B sero-strip showed to be highly sensitive and specific in the detection of antibodies (IgG) against saliva. In the future, this test can be employed during large-scale epidemiological studies of CanL in the Mediterranean area to evaluate the effectiveness of vector control programs. Author summary The sand fly is the basic principle vector of in the Mediterranean area. Since standard serological methods are impractical and time-consuming in field conditions, we propose the rSP03B sero-stripa quick test that can be immediately applied to screen large cohorts of dogs for the presence of anti-antibodies. Our test is the 1st rapid test in the field of vector exposure, it is highly sensitive and specific and shown to be a valid replacement Rbin-1 for standard serological assays. In addition, this test could be used as an evaluation tool for vector control programs. Introduction Canine leishmaniasis (CanL) is definitely a common zoonotic disease present in several countries in Latin-America, Europe and Asia [1,2]. It is a severe multi-systemic disease of dogs caused by the protozoan parasite is the most important. During the bite, the sand take flight injects saliva comprising a cocktail of bio-active molecules with anti-hemostatic, anti-inflammatory and immune-modulatory activities into the sponsor skin (examined in [10]). These molecules facilitate the blood-feeding process of the sand fly and result in a humoral immune response in the sponsor. It is well-known that the amount of sponsor anti-saliva IgG antibodies (Abs) correlates with the level of exposure to sand flies [11,12]. Furthermore, earlier studies showed a definite fluctuation of the Ab response during longitudinal Rabbit Polyclonal to MCL1 sampling of dogs over two transmission seasons [26], suggesting that proteins present in sand fly saliva can be a useful tool to evaluate the effectiveness of vector control programs. For example, earlier studies on mosquitoes [13C15] and triatomine insects [16] have shown that a reduction in vector denseness observed after the implementation of insecticide treated nets (ITNs) correlates with a reduction in anti-vector salivary Ab-response. With regard to sand flies, only one study performed in India and Nepal measured anti-Abs to evaluate the use of ITNs [17]. Performing large-scale serological studies to detect sponsor exposure to sand take flight bites was limited in the past due to the fact that dissecting large amounts of sand take flight salivary glands is definitely a demanding and labour-intensive process. Besides, the use of whole SGH is subject to protein content variability, dependent on the age of the sand fly at the time of dissection [18] and might antigenically cross-react with taxonomically closely related sand fly Rbin-1 varieties (examined in [19]). Consequently, using specific antigenic recombinant sand fly salivary proteins as a replacement to the use of whole SGH has gained more attention [20C23]. Previously the specific antibody response (IgG) against the salivary protein SP03B from was proposed like a valid exposure marker across areas endemic for CanL [22]. This study demonstrated the presence of related antigenic epitopes in the recombinant SP03B protein compared to its native form, and indicated a substantial antigenic cross-reactivity amongst populations from Campania, Umbria and the metropolitan Lisbon region [22]. The.