In the present study, we described an improved method for the development and application of the RT-LAMP assay for the detection of DENV infection in freshly obtained dengue-suspected patient samples in actual clinical laboratory setting of a hospital in dengue endemic environment.
All clinical laboratories in Hawaii were asked to report any requests for dengue diagnostic testing and to forward aliquots of serum samples obtained for dengue testing to the HDOH State Laboratories Division.
Recent data obtained with Dengue virus showed that carrageenan may interfere not only with adsorption of virus to cells but also block the fusion event leading to uncoating of the nucleocapsid [36].
Using DENV-1 isolates from Sri Lanka obtained from dengue cases in 1983 , 1984 , 19972003, and 2004 (7 ) and representative DENV-1 sequences for the 4 genotypes, we constructed a phylogenetic tree by using MEGA5 software (10 ).
The clinical applicability of a single-tube RT-LAMP assay for detection of DENV RNA was assessed in a total of 305 serum samples freshly obtained from dengue-suspected patients.
Sera samples were obtained from positive dengue patients in the acute phase of infection during the dengue epidemical outbreaks of years 2006 to 2008.
The satisfactory results obtained for the dengue dataset encourages the application of GP to model relatively small datasets issued from a single laboratory.
The patient serum samples were obtained from a dengue cohort developed at the Virology and Experimental Therapy Laboratory (LAVITE), of the Aggeu Magalhães Research Center Recife, Brazil described elsewhere [23].
Informed written consent, approved by Fundação Oswaldo Cruz Ethical Committee under Nr#111/00, was obtained from all dengue patients prior to blood draw.
