Detección y cuantificación de virus dengue 2 en lisado celular y plasma de niños por qPCR en tiempo real usando un estuche comercial y el equipo EcoTM System-Illumina
Detección y cuantificación de virus dengue 2 en lisado celular y plasma de niños por qPCR en tiempo real usando un estuche comercial y el equipo EcoTM System-Illumina
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Methods for Dengue virus (DENV) diagnosis in endemic areas are greatly needed. One of them is the real-time polymerase chain reaction (qPCR) that also enables to quantitate the viral genome. Kits of qPCR for DENV are expensive and restrict their use to a small number of qPCR devices, which limits the application of the technique. Here, we evaluated the performance of a commercial kit of qPCR to DENV-2 detection on a locally available qPCR device (EcoTM System, Illumina), not cited by the kit manufacturer.VERO-76 cells lysate and plasma from children, both with confirmed ongoing DENV-2 infection, were evaluated. As specificity control, cell lysates and plasma from children infected with DENV-1, and uninfected lysate, were also included. The reactions were simultaneously evaluated in an Applied Bio systems 7300 device. The standard curve generated by EcoTM was robust (R2= 0.99) with low variability in the replicates (<10%). The reaction efficiency was high (88.8%) and signal was only obtained in lysates and plasma infected with DENV-2. There was a strong positive correlation (R2= 1.0, P= 0.0028) between the number of copies of viral RNA in the samples detected by both qPCR devices. Thus, the use of the evaluate kit for detection of DENV-2 here tested can be extended to EcoTM. With this work, technological capacity for DENV study in an endemic zone is greatly strengthened.
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