Based on viral isolation, HAI, and an in-house ELISA, 2 patients were diagnosed with primary dengue (squares), 22 were diagnosed with secondary dengue (triangles), and 11 had no evidence of dengue infection (circles)

Based on viral isolation, HAI, and an in-house ELISA, 2 patients were diagnosed with primary dengue (squares), 22 were diagnosed with secondary dengue (triangles), and 11 had no evidence of dengue infection (circles). donors (n= 17) showed elevation of salivary antidengue antibody (100% specificity). Salivary IgG levels correlated well with serum HAI titer (r= 0.78), and salivary IgG levels could be used to distinguish between primary- and secondary-dengue virus infections. In terms of morbidity, mortality, and economic costs, dengue is the most important mosquito-borne disease in the world, with an estimated 100 million cases annually (13). Initial infection with one of the four serotypes of dengue virus (primary-dengue virus infection) may lead to dengue fever, which is a self-limiting, febrile disease with a low mortality rate, while reinfection with a different dengue serotype (anamnestic or secondary-dengue virus infection) may lead to more-serious forms of the disease (e.g., dengue hemorrhagic fever or dengue shock syndrome) (1,9,14). Recently, commercial tests have been described for the detection of anti-dengue immunoglobulin M (IgM) and IgG antibodies in serum (2,11,12,21,23). Potential AAF-CMK problems with the use of serum include the requirement of consent and cooperation of the patient, which is often unavailable due to social or religious reasons, the need for a trained venipuncturist and AAF-CMK the need to separate serum before testing, and the difficulty and added risk of venipuncture in children, the group most commonly affected by dengue in areas where infection is endemic. Most body fluids contain antibodies, although at much lower levels than those in blood. Thus, these sources of antibody are unsuitable as diagnostic specimens, in spite of the obvious advantages and convenience of samples such Rabbit polyclonal to PRKAA1 as saliva. Salivary antibodies have been reported to be useful for the diagnosis of a number of infections, including AIDS, leptospirosis, measles, mumps, hepatitis A and B, and rubella (36,1517). In this study AAF-CMK we examined the ability of the PanBio Dengue Duo enzyme-linked immunosorbent assay (ELISA) to detect both IgM and IgG antibodies to dengue with saliva samples. Sera and saliva samples were collected prospectively from patients presenting at the Kamphaeng Phet Provincial Hospital in northern Thailand. Saliva was collected by using a commercially available collection device (Omni-Sal; Salivary Diagnostic Systems, Singapore). This device dilutes saliva twofold in the buffer provided. After collection, saliva was stored at 80C until assayed blindly by the Dengue Duo ELISA. Diagnosis was based on assay of blood or sera by using in-house ELISA, hemagglutination inhibition assay (HAI), or viral isolation performed at the Armed Forces Research Institute of Medical Sciences (AFRIMS) in Bangkok, Thailand (8,21). Of the 35 patients from Thailand enrolled in the study, 2 had primary dengue, 22 had secondary dengue, and 11 had no laboratory evidence of dengue infection despite the presence of clinical symptoms compatible with dengue fever. Saliva was also collected from 17 healthy Australian laboratory staff members. The Dengue Duo ELISA has been shown to be useful in the diagnosis of dengue infection with sera (2,12). It detects IgM and IgG separately by a capture assay format and was performed by the procedure recommended by the manufacturer (2), except that saliva was diluted 1:2 in the assay diluent provided before the addition of 100 l to each well of the assay plate (final dilution, 1:4). Positive, negative, and calibrator control sera used in the kit were also run alongside the saliva samples, though these were diluted 1:100 in the diluent provided. Results were AAF-CMK expressed as the ratio of the absorbance in test samples divided by AAF-CMK the absorbance of the calibrator sera. A ratio of 0.6 was found to give the best distinction between dengue infection and other conditions. A positive sample was defined as having a sample/calibrator absorbance ratio of 0.6, and a negative sample was defined as having a sample/calibrator absorbance ratio of <0.6. Dengue virus infection was characterized by the elevation of either.