Blocked nitrocellulose was placed on an 80 mm backing card (Lohmann). A 22 mm Ahlstrom 320 wicking pad (Ahlstrom-Munksjo Oyj, Finland) was placed on top of the downstream edge of the nitrocellulose with a 5 mm overlap between the two materials. The sprayed and blocked conjugate pad was placed on top of the upstream edge of the nitrocellulose with a 2 mm overlap. Finally, the sample pad is placed on top of the upstream edge of the conjugate pad with a 2 mM overlap. Strips were cut to a 5 mm width using a Kinematic Matrix guillotine cutter (Kinematic Automation, Inc., Twain Harte, CA, USA). Strips were then placed in individual injection molded cassettes and stored in desiccator until use.