Abstract
BackgroundDengue virus (DENV) infection is one of the biggest challenges for human health in the world. In addition, a secondary DENV infection sometimes causes dengue hemorrhagic fever (DHF), which frequently leads to death. For this reason, accurate diagnosis record management is useful for prediction of DHF. Therefore, the demand for DENV rapid diagnosis tests (RDTs) is increasing because these tests are easy and rapid to use. However, commercially available RDTs often show low sensitivity for DENV and cross-reactivity against other flaviviruses, especially Zika virus (ZIKV).MethodsWe developed two types of novel DENV non-structural protein 1 (NS1) detection RDTs, designated TKK-1st and TKK-2nd kits. Specificities of the monoclonal antibodies (MAbs) used in these kits were confirmed by enzyme-linked immuno-sorbent assay (ELISA), dot blot, and western blot using recombinant NS1 proteins and synthetic peptides. For evaluation of sensitivity, specificity, and cross-reactivity of the novel DENV NS1 RDTs, we first used cultured DENV and other flaviviruses, ZIKV and Japanese encephalitis virus (JEV). We then used clinical specimens obtained in Bangladesh in 2017 for further evaluation of kit sensitivity and specificity in comparison with commercially available RDTs. In addition, RNA extracted from sera were used for viral genome sequencing and genotyping.ResultsEpitopes of three out of four MAbs used in the two novel RDTs were located in amino acid positions 100 to 122 in the NS1 protein, a region that shows low levels of homology with other flaviviruses. Our new kits showed high levels of sensitivity against various serotypes and genotypes of DENV and exhibited high levels of specificity without cross-reactivity against ZIKV and JEV. In clinical specimens, our RDTs showed sensitivities of 96.0% (145/151, TKK-1st kit) and 96.7% (146/151, TKK-2nd kit), and specificities of 98.0% (98/100, TKK-1st kit and TKK-2nd kit). On the other hand, in the case of the commercially available SD Bioline RDT, sensitivity was 83.4% (126/151) and specificity was 99.0% (99/100) against the same clinical specimens.ConclusionsOur novel DENV NS1-targeting RDTs demonstrated high levels of sensitivity and lacked cross-reactivity against ZIKV and JEV compared with commercially available RDTs.
Highlights
Dengue virus (DENV) infection is one of the biggest challenges for human health in the world
After washing three times with phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA), the AuNP-conjugated monoclonal antibodies (MAbs) was diluted to an optical density of 2.5 and impregnated into glass fibers (Millipore, Billerica, MA), which were dried in an incubator
Our novel DENV non-structural protein 1 (NS1)-targeting rapid detection test (RDT) demonstrated high levels of sensitivity and lacked cross-reactivity against Zika virus (ZIKV) and Japanese encephalitis virus (JEV) compared with commercially available RDTs
Summary
Dengue virus (DENV) infection is one of the biggest challenges for human health in the world. A secondary DENV infection sometimes causes dengue hemorrhagic fever (DHF), which frequently leads to death. For this reason, accurate diagnosis record management is useful for prediction of DHF. Primary DENV infection causes DF, whereas a small but non-negligible proportion of patients with secondary infection manifest the more severe symptoms of dengue hemorrhagic fever (DHF), which can cause death [8]. It is important to obtain accurate patient records regarding previous DENV infection to predict the severity of ensuing disease, permitting clinical risk management of DHF. Easy access to rapid diagnosis would facilitate the generation of accurate records of DENV infection
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