Japanese encephalitis (JE) is an emerging mosquito-borne zoonotic flaviviral disease. The present study was undertaken with the objective to develop TaqMan real-time reverse-transcription polymerase chain reaction (RT-PCR) assay for rapid detection and quantification of Japanese encephalitis virus (JEV) in swine blood and mosquito vectors. The amplification of envelope (E) gene was targeted by designing gene-specific MGB TaqMan fluorescent probe along with the primers. The best performance in terms of sensitivity was achieved by standardized TaqMan real-time RT-PCR with a detection limit of 2.8 copies/reaction and it was found to be 4-log more sensitive than conventional RT-PCR. The applicability of the standardized TaqMan assay was evaluated by screening representative sets of field swine blood samples and mosquito pools for JEV. The viral load ranged between 3.32 × 107–4.2 × 102 copies/ml of swine blood samples, and 5.7 × 109–1.3 × 102 copies/pool of mosquitoes. The standardized assay which is highly sensitive, specific and rapid would aid in screening sentinel swine and mosquitoes under JEV surveillance programs for effective prevention and control of disease in human beings.
Japanese encephalitis (JE) is a re-emerging mosquito-borne zoonotic flaviviral disease. Swine sero-convert 2–3 weeks before infection occurs in humans and thus serves as a suitable sentinel for JE surveillance and outbreak prediction in human population. The present study was conducted with the objective of developing a lateral flow assay (LFA) for detecting JEV antibodies in swine sera. Three different formats were tried using recombinant NS1 protein as antigen in order to select the best format. In format I, gold nanoparticles were conjugated with antigen followed by spotting of antigen on NCM as test line and anti-antigen IgG on NCM as control line. In format II, gold nanoparticles were conjugated with antigen followed by spotting of staphylococcal protein A as test line and anti-antigen IgG as control line. Format III used gold nanoparticles conjugated with goat anti-pig IgG followed by spotting of antigen as test line and pig IgG as control line. Amongst the three formats, format II was found to be superior with 100% relative diagnostic sensitivity and 100% relative diagnostic specificity during monsoon and post-monsoon period. A panel of 500 field swine serum samples was tested using format II which revealed sero-positivity of 15.6%, and the format was found suitable to screen swine serum samples during monsoon and post-monsoon period.
Japanese encephalitis virus (JEV) causes ~100,000 clinical cases and 25,000 deaths annually worldwide, mainly in Southeast Asia and the Western Pacific and mostly in children. JEV is transmitted to humans through the bite of mosquitoes that have fed on competent hosts. Abiotic factors, such as seasonal rainfall, influence transmission. Transmission models have an important role in understanding disease dynamics and developing prevention and control strategies to limit the impact of infectious diseases. Our goal was to investigate how transmission models capture JEV infection dynamics and their role in predicting and controlling infection. This was achieved by identifying published JEV transmission models, describing their features and identifying their limitations, to guide future modelling. A Preferred Reporting Items for Systematic Reviews and Meta‐Analyses Extension for Scoping Reviews (PRISMA‐ScR)‐guided scoping review of peer‐reviewed JEV transmission models was conducted. Databases searched included PubMed, ProQuest, Scopus, Web of Science and Google Scholar. Of the 881 full‐text papers available in English, 29 were eligible for data extraction. Publication year ranged from 1975 to 2023. The median number of host populations represented in each model was 3 (range: 1–8; usually humans, mosquitoes and pigs). Most (72% [ n = 21]) models were deterministic, using ordinary differential equations to describe transmission. Ten models were applied (representing a real JEV transmission setting) and validated with field data, while the remaining 19 models were theoretical. In the applied models, data from only a small proportion of countries in Southeast Asia and the Western Pacific were used. Limitations included gaps in knowledge of local JEV epidemiology, vector attributes and the impact of prevention and control strategies, along with a lack of model validation with field data. The lack and limitations of models highlight that further research to understand JEV epidemiology is needed and that there is opportunity to develop and implement applied models to improve control strategies for at‐risk populations of animals and humans.