Diagn Microbiol Infect Dis

Early diagnosis of leptospirosis in humans is critical with regard to initiation of appropriate treatment; however, the gold standard serological test cannot detect antibodies until nearly a week after symptom onset. PCR has been shown to be sensitive and specific in the early phase of leptospirosis. Previously, we developed and validated a TaqMan PCR assay targeting lipL32. We reoptimized and validated this assay using PerfeCTa\uae qPCR ToughMix\uae, Low ROX\u2122 (Quanta Biosciences, Gaithersburg, MD, USA). For optimization with the new mix, the final primer concentrations were increased from 0.5 \u3bcmol/L to 0.9 \u3bcmol/L compared to our previous assay, and the probe concentration increased from 0.1 \u3bcmol/L to 0.125 \u3bcmol/L. This newly optimized assay resulted in a lower limit of detection and increased diagnostic sensitivity. Here, we present the performance data of the improved assay and describe several clinical cases that were initially negative but tested positive using the optimized assay.CC999999/Intramural CDC HHS/United States2019-04-08T00:00:00Z25912810PMC64524406127vault:3186