Molecular diagnosis and transmission of dengue in Sri Lanka

Abstract

BACKGROUND: Dengue virus is an important vector-borne viral infection in Sri Lanka. Laboratory confirmation of suspected dengue cases is important for over/under estimation of cases. Early rapid diagnosis of dengue viral infection helps monitoring the disease, hospital admission when necessary and reduces case fatality. Detection of dengue viruses in mosquitoes is useful for studies on transmission of dengue virus. Study on risk factors for dengue is useful to understand spatial and temporal dynamics of transmission the disease. METHODOLOGY: A novel molecular-based assay; Reverse Transcription-Polymerase Chain Reaction-based Liquid Hybridization (RT-PCR-LH) was developed and validated for detection of dengue virus in clinical and mosquito specimens. Severity of dengue with circulating serotype was also analyzed. Wild-caught mosquito samples were collected from 236 dengue case-reported stations during outbreaks and a hot-spot during a period of 31 months. Epidemiological, environmental and entomological and other possible risk factors affecting transmission of dengue were analyzed. RESULTS: As an early (<5 days of fever) laboratory diagnostic method for dengue virus, the novel assay had 100% and 46% sensitivity for detection of confirmed and suspected dengue patients respectively. The assay developed in this study was found to be more sensitive than the other diagnostic techniques for early definitive laboratory diagnosis of dengue infection. Patients with definitive dengue correlated only with few signs and symptoms, indicating that laboratory confirmation is critical to avoid over estimation. A high sensitivity of 2 fluorescent focus unit of dengue virus/reaction was achieved and the assay was highly specific for dengue virus. The assay could detect dengue virus in 7% of field-caught Aedes albopictus specimens. A high density of Ae. albopictus was also associated with the dengue case-reported stations/hot-spots. Both vector species were susceptible to the 4 dengue serotypes under the laboratory conditions and DEN-3 under the field conditions. Geographical Information System (GIS) based risk mapping and database including epidemiological, climatic condition and entomological surveillance information were developed for the hot-spot in Kurunegala during the period of 2000-2003, which can be used as an early warning system. In depth study on socio-economic and other related factors affecting transmission of dengue was studied in the District of Gampaha. CONCLUSION: Ae. albopictus acts as an important vector of transmission of dengue in some urban and semi-urban areas. GIS-based risk maps developed are important to predict impending epidemics so that limited resources could be utilized in a cost effective manner to control the disease. Some socio-economic factors directly affecting transmission of dengue.