Rational design and characterization of a lateral flow assay for canine C-reactive protein in wound exudate.

C-reactive protein levels may have clinical value in monitoring different phases of healing and in identifying possible states of infection. As a critical step to further investigate this potential connection, we have demonstrated a lateral flow assay (LFA) for canine CRP level assessment in wound exudate that could be used as a tool in the veterinary clinic setting. In the rational design of our cCRP LFA, we have characterized LFA performance for sequential delivery mode vs. the more common premixed delivery mode using several metrics including dynamic range, sensitivity, limit of detection, and time to result. Although the sequential mode assay results indicated modestly improved signal (3-14%) and limit of detection (in the low ng/mL range for both) for this set of cCRP immunoassay reagents, the premixed mode assay's shorter run time with one less delivery step was chosen for use in this application in which analyte levels are substantially elevated. We have defined a straightforward wound exudate processing procedure that includes centrifugation to extract exudate from canine patient bandages, and subsequent sample dilution for cCRP quantification by our LFA. And, we have demonstrated that our cCRP LFA provides comparable cCRP concentrations to that of gold-standard ELISA performed on the same clinical wound exudate, and serum/plasma samples. Finally, we have highlighted some next steps in the assessment of cCRP as a biomarker for wound healing and infection.