Design of Multiplex Lateral Flow Tests: A Case Study for Simultaneous Detection of Three Antibiotics

Abstract

The presented study is focused on the impact of binding zones locations at immunochromatographic test strips into analytical parameters of multiplex lateral flow assay. Due to non-equilibrium conditions for such assays the duration of immune reactions influences significantly on analytical parameters, and the integration of several analytes into one multiplex strip may cause essential decrease of sensitivity. To choose the best location of binding zones, we have tested reactants for immunochromatographic assays of lincomycin, chloramphenicol, and tetracycline. The influence of the distance to the binding zones on the intensity of coloration and limit of detection (LOD) was rather different. Basing on the obtained data, the best order of binding zones was chosen. In comparison with non-optimal location the LODs were 5-10 fold improved. The final assay provides LODs 0.4, 0.4 and 1.0 ng/mL for lincomycin, chloramphenicol, and tetracycline, respectively. The proposed approach can be applied for multiassays of other analytes.

Hyphenating paper-based biosensors with smartphones

“…This means that the binding interactions at the first test line can subsequently interfere with downstream test lines by slowing the liquid flow across the membrane, or even blocking other antibody/analyte complexes from proceeding past. In this case, careful consideration must be given to the compatibility of different antigens on the same test, the order the lines are printed [57], or how the lines are printed [52].…”