Development of LAMP for EBV and cells-to-LAMP Assay. (A) Top: gel electrophoresis results of EBER LAMP reaction run for 30 minutes in a standard thermal cycler. Bottom: in-tube turbidity immediately after reaction. Lane/tube 0: 100bp DNA ladder; lane/tube 1: 480 copies EBER plasmid DNA control; lane/tube 2: Namalwa (EBV+) DNA; lane/tube 3: Raji (EBV+) DNA; lane/tube 4: BC-3 (EBV-) DNA; lane/tube 5: Ramos (EBV-) DNA; lane/tube 6: water. (B) Amplification of serially diluted plasmid DNA amplified with EBER LAMP in real-time. Fifty of 51 samples containing 90 copies per reaction were amplified before the 30-minute threshold cutoff. (C) Amplification threshold results of purified DNA from cells with Qiagen DNeasy and the cells-to-LAMP procedure, no significant difference detected, P = .2222 by Mann-Whitney U test. Each sample contained the same number of cells (1 million cells) in the same final volume across assays. Data show combined results of 4 independent experiments containing ≥3 replicates in each group shown together. (D) Stability assessment of amplification from cell lines stored in cells-to-LAMP solution at room temperature (∼25°C), 4°C, or −20°C at 0, 4, or 8 weeks after extraction. The EBV– control Ramos was also evaluated but is not shown because no amplification was seen before our 30-minute threshold. (E) Amplification threshold times at high (10 million cells) and low (50 cells) loads in a conventional real-time thermal cycler were compared with the TINY system. (F) Cells-to-LAMP amplification times from 1 million cells from EBV+ and EBV- cell lines in TINY show specificity. The EBV– cell lines did not amplify (NA).