Several previous studies have utilized SYBR Green for HIV-1 diagnosis and are reviewed by Espy et al. [57] The efficiency of this dye-based assay is also comparable with the currently available diagnostic assays on HIV. Aside from use in HIV diagnosis and quantification, SYBR Green has been widely used to detect and quantify diverse human pathogens. [58] [59] [60] [61] [62] [63] [64] [65] The data in these reports strongly support the utility and reliability of SYBR Green for detection of specific PCR amplicons above the background. The use of degenerate LTR primers in the rtLC DBS assay allows for a wide range of genotype inclusivity.