ELISA as an alternative tool for epidemiological surveillance for dengue in mosquitoes: a report from Thailand.

BACKGROUND & OBJECTIVES: Dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are the re-emerging infectious diseases caused by the four serotypes of dengue (DEN) virus, type 1 to 4, belonging to the family Flaviviridae and genus Flavivirus. In the absence of a safe and effective mass immunisation, the prevention and control of dengue outbreaks depend upon the surveillance of cases and mosquito vector. The aim of this work is to test enzyme-linked immunosorbent assay (ELISA) tool for the virological surveillance of dengue. METHODS: Virus-infected Aedes mosquitoes were collected from the field in order to serve as an early warning monitoring tool for dengue outbreaks. In a prospective field study conducted from April to September 2000, female adult Aedes mosquitoes were caught from selected dengue-sensitive area in Chombung district, Ratchaburi province and assayed by ELISA. RESULT: Approximately 18.3% were found positive for dengue virus. CONCLUSION: This can imply that ELISA can be an alternative tool for epidemiological surveillance for dengue in mosquitoes.

Border and imported bancroftian filariases: baseline seroprevalence in sentinel populations exposed to infections with Wuchereria bancrofti and concomitant HIV at the start of diethylcarbamazine mass treatment in Thailand.

Border bancroftian filariasis caused by Wuchereria bancrofti nocturnally subperiodic mainly exists in Karens residing alongside the Thailand-Myanmar border. Imported bancroftian filariasis caused by W. bancrofti nocturnally periodic mainly exists in cross-border Myanmar migrants. We analyzed seroprevalence data based on W. bancrofti adult worm antigen (Ag) loads and human immunodeficiency virus (HIV) immunoglobulins in the sentinel population samples which were studied prior to the start of the diethylcarbamazine (DEC) mass treatment phase in the PELF during fiscal years 2002-2006. In the Karens, the cumulative infection prevalence (36.8% serological antigen positivity or SAP) was specific for age (p < 0.001) but universal for gender (p = 0.77). The infection intensity (median Ag load = 60,827 antigen units or AU/ml) was specific for age (p = 0.031) and for males (p = 0.016). In the Myanmars, infection prevalence (24.0% SAP) was universal for age (p = 0.961) and for gender (p = 0.676). The infection intensity (median Ag load = 19,068 AU/ml) was universal for age (p = 0.433) but specific for females (p = 0.027). Overall, the Ag loads between the groups were significantly different (p = 0.014). In analysis of concomitant HIV and W. bancrofti infections, 7 (3.2%) Myanmars infected with HIV 1 and 3 (5.7%) with concomitant infections, subjected to biannual DEC treatment with 300 mg oral-dose FILADEC, were prevalent. The antigenemia clearance in the concomitant infections (r = -0.732, p = 0.039) as well as in the single W. bancrofti infection (r = -0.781, p = 0.022) was correlated with time required to clear antigenemias. We reemphasize that W. bancrofti adult worm Ag loads in the sentinel population samples would be beneficial for the PELF's implementers at the provincial level to probe the disease burdens in target areas and to evaluate and monitor the DEC treatment efficacy and effectiveness in those sentinel populations, including those with concomitant HIV eligible for the DEC mass treatment phase in the PELF.

The geographic information system as an epidemiological tool in the surveillance of dengue virus-infected Aedes mosquitos.

A Geographic Information System (GIS) was used as analysis tool to study the spatial distribution of dengue virus-infected Aedes mosquitos in Thailand. Global Positioning System (GPS) instruments were used to map villages involved in dengue epidemiological studies in Ratchaburi Province, Thailand. Differentially processed GPS data, with a spatial resolution of approximately 1 meter, were incorporated into a GIS for analysis and mapping. Databases associated with a village GIS included village number, Aedes aegypti populations, and test results. Epidemiological surveillance for dengue infection through the detection of the dengue virus type(s) infecting Aedes mosquitos during epidemic periods constitutes a reliable sentinel system for dengue outbreaks. Various techniques were applied including: enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent assay (IFA), and reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the virologic surveillance of the type-specific detection of dengue viruses in artificially infected and in field-caught adult Aedes mosquitos. In laboratory experiments, all assays showed sufficient sensitively to detect one virus infected mosquito and the rapid RT-PCR clearly showed serotype-specificity with very high detection sensitivity. In the field study conducted from April to September 2000, female adult Aedes mosquitos were collected from selected dengue-sensitive areas in Chom Bung district, Ratchaburi Province and assayed by ELISA, IFA and RT-PCR with 18.3% (44/240), 28.98% (20/69) and 15% (3/20) positive for dengue virus, respectively. Geographic distribution of the virus-infected Aedes mosquitos and household locations were demonstrated by the GPS and the GIS. The development of disease mapping data coupled with RT-PCR laboratory-based surveillance of dengue virus infection can successfully serve as epidemiologic tools in an early warning system for dengue hemorrhagic fever (DHF) epidemics.

The epidemiology of HIV-1 subtypes in infected patients from northeastern Thailand.

In total, 117 HIV-1 infected patients from several provinces in Northeastern Thailand were analysed. All blood samples collected from individuals were confirmed by EIA and Western blot and partially by HIV-1 gag-, pol- and env-PCR. By serotyping with a V3-peptide ELISA, 108 (92.3%) of the sera samples belonged to subtype E, 9 (7.7%) were serotype B. For 10 Thai HIV-1 infections, the serotype and genotype were determined. The genotype was determined by phylogenetic analysis of directly sequenced PCR amplicons, 8 were subtype E, 2 subtype B. For these patients the serotype did correlate with the genotype. Tracing back the origin of Thai patients, it seems that most were infected within early years of the epidemic and the Thai subtype B infected patients have been imported directly from foreign countries via sexual contact. The findings suggest there are two district subtypes in Thailand with the majority being subtype E. The relatively high prevalence of subtype B in Northeastern Thailand may be due to the increasing intermix of the two strains (subtypes E and B) and the migration for employment from foreign countries. This may lead to public health concerns regarding surveillance of HIV-1 subtypes and the regulation of potentially infected workers returning from abroad to the country.