Superscript 3 platinum one step quantitative rt pcr system with rox kit

During the last dengue epidemic of 2019–2021, 3228 available sera from dengue cases confirmed by the IPG, the CHUG, and the CHUM were used. Venous blood samples (5 mL) from symptomatic patients were collected by several laboratories in the frame of their diagnostic activities and stored at 4 °C during the transportation to the laboratories.
In Guadeloupe, serum samples from CHUG and IPG were first submitted to nucleic acid extraction using the NucleoSpin^® 96 Virus kit (Macherey Nagel, Duren, Germany) at IPG, following the manufacturer’s instructions, and then screened for presence of DENV by real-time RT-PCR using primers previously described [25 (link)]. If samples were DENV-positive, serotyping was conducted using specific real-time RT-PCR primers [26 (link),27 ]. The real-time RT-PCRs were performed using Superscript^® III Platinum One step Quantitative RT-PCR System with ROX kit (Invitrogen, Carlsbad, CA, USA) and the 7500 Real-Time PCR (Applied Biosystems, Waltham, MA, USA).
In Martinique, nucleic acids were extracted at CHUM from plasma using the NucliSens easyMag automated system (BioMérieux, Craponne, France). Samples were then screened by real-time RT-PCR using the Simplexa^TM Dengue Kit (DiaSorin Molecular, Cypress, CA, USA) for virus detection and serotype determination.