Pilot Investigation into the Ruggedized Advanced Pathogen Detection Device (R.A.P.I.D.) for Dengue Detection in East Timor

摘要

Polymerase chain reaction (PCR) assays for detection of vector transmitted virus infections in humans designed for conventional PCR thermal cyclers were modified for real-time PCR using a LightCycler. Two reporter fluorochromes, the double stranded DNA specific dye SYBR Green I, and 6-hydroxy fluorescein conjugated to TaqMan DNA probes, were employed to detect positive reactions. Following optimisation of the assays for real-time PCR, portable real-time PCR equipment, the Ruggedized Advanced Pathogen Detection Device (R.A. P.I.D.(trademark)), was deployed to East Timor and a field trial was conducted from November 2001 until January 2002. Blood samples from a total of 143 army personnel were screened with the R.A.P.I.D. for virus infections. These included 33 samples from patients showing clinical symptoms. Results obtained with the R. A.P.I.D. were compared to results from serological testing and cell culture analysis.