Journal: Viruses

Article Title: QuickFit: A High-Throughput RT-qPCR-Based Assay to Quantify Viral Growth and Fitness In Vitro

doi: 10.3390/v16081320

Figure Lengend Snippet: Analysis and modeling of HIV in vitro replication growth curves as measured by p24 ELISA and QuickFit RT-qPCR. Both p24 ELISA (left) and QuickFit RT-qPCR (right) were performed with samples obtained from HIV REJO.c -infected CD4 + T cells. ( A ) Determined raw p24 concentrations (pg/mL) and ( B ) viral loads (GC/mL) are shown. A total of 16 samples at five different time points were evaluated, and matching light–dark colors represent experimental duplicates. ( C , D ) Samples were subjected to inclusion and exclusion criteria, and the filtered data were used to determine growth rates and carrying capacities by running an NMLE modeling with a half-maximal equation in the MonolixSuite 2021R software. ( E , F ) Viral growth (p24 concentration or GC/mL) was calculated by normalizing the initial value on day 0 to 1.00 pg/mL or 1 × 10 5 GC/mL and subsequently interpolating the remaining values from days 1 to 4 based on the fit curve model. Overlaid is the mean growth rate and the 95% confidence interval. The r value represents the average growth rate determined for all included samples by each assay. The dotted lines represent the limit of detection for each assay.

Article Snippet: After filtering, viral growth indicator data were analyzed using the non-linear mixed-effect (NLME) modeling software MonolixSuite 2021R (Lixoft SAS, Antony, France) [ , ].

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Infection, Software, Concentration Assay