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  • For molecular diagnostics, one-step qRT-PCR was performed according to the Center of Disease Control (CDC) protocol using primers and probe designed for Caribbean lineage (Asian genotype) [29]: 1) we used the primers CHIK 856 5’-ACCATCGGTGTTCCATCTAAAG-3’ and CHIK 962c 5’-GCCTGGGCTCATCGTTATT-3’ and the TaqMan Probe CHIK 908 5’-FAM- ACAGTGGTTTCGTGTGAGGGCTAC-NFQ-3´; 2) the qRT-PCR was prepared with the QIAGEN QuantiTect Probe RT-PCR kit® (Qiagen, Venlo, Netherlands) in accordance with the manufacturer’s protocol, adding 10μL of RNA sample per reaction in duplicate; and 3) the samples were run in the 7500 Fast Real Time PCR System (Applied Biosystems, Foster City, CA).
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