Multiple test zones for improved detection performance in lateral flow assays

Lateral flow assays (LFAs) have found applications in clinical diagnostics, veterinary medicine, food safety, environmental monitoring, bio-defence, drug abuse, etc. However, conventional LFAs with single test zone for one analyte (sLFA) show qualitative, positive/negative results (that are usually not so accurate for decision-making) and suffer from the hook effect (that easily induces false negative results). In this study, a modified LFA with multiple test zones for one analyte (mLFA) is developed for detection of nucleic acid showing two methods to interpret assay results. One is the sum of detection signals of test zones, which exhibits a stable and broader detection range compared with sLFA. The other is the pattern of test zones that is presented is correlated to the concentration of analyte, facilitating semi-quantitative detection without the aid of a reader or analyzer.