Optimization of Fecal Calprotectin Assay for Pig Samples
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Fecal calprotectin is a noninvasive marker of gut inflammation and has been widely utilizedin human gastrointestinal diagnostics. This marker, however, has not been extensively utilized inporcine samples. The aim of this study was to optimize a protocol for the extraction of porcinefecal calprotectin, and to the best of our knowledge this is the first study to be conducted inthis regard. Freshly collected swine fecal samples were used in this study. We determined thevariability of three commercial ELISA assays in the recovery of porcine fecal calprotectin. Wefurther studied the effect of dilution factor and roller shaker homogenization on the yield ofcalprotectin from swine fecal samples. Calprotectin recovery was significantly different(p<0.05)across the three commercial assays with MBS033848 having a greater recovery compared toDAEF-012 and calprest. Fecal calprotectin yield increased with an increase in dilution factorwith maximum recovery at 1:250. Furthermore, homogenization of fecal sample extracts using aroller shaker for tubes for 30 min led to a 30.75% relative increase in calprotectin yield. Further increase in shaking time(at 60 min) led to a reduced calprotectin recovery. Calprotectinrecovery ratio was 130.8% and 101.4% at 30 min and 60 min homogenization respectively. Inour conclusion, we observed that various factors affect the recovery of porcine fecal calprotectin,and therefore the researcher should double check certain parameters in regard to the type of kit,the dilution factor and homogenization time if reliable and reproducible results are to beobtained. Results of the present study provide useful information on a non-invasive protocol toveterinarians and researchers in examining and monitoring swine gut healthusing the fecalcalprotectin.
Abstract
Fecal calprotectin is a noninvasive marker of gut inflammation and has been widely utilizedin human gastrointestinal diagnostics. This marker, however, has not been extensively utilized inporcine samples. The aim of this study was to optimize a protocol for the extraction of porcinefecal calprotectin, and to the best of our knowledge this is the first study to be conducted inthis regard. Freshly collected swine fecal samples were used in this study. We determined thevariability of three commercial ELISA assays in the recovery of porcine fecal calprotectin. Wefurther studied the effect of dilution factor and roller shaker homogenization on the yield ofcalprotectin from swine fecal samples. Calprotectin recovery was significantly different(p<0.05)across the three commercial assays with MBS033848 having a greater recovery compared toDAEF-012 and calprest. Fecal calprotectin yield increased with an increase in dilution factorwith maximum recovery at 1:250. Furthermore, homogenization of fecal sample extracts using aroller shaker for tubes for 30 min led to a 30.75% relative increase in calprotectin yield. Further increase in shaking time(at 60 min) led to a reduced calprotectin recovery. Calprotectinrecovery ratio was 130.8% and 101.4% at 30 min and 60 min homogenization respectively. Inour conclusion, we observed that various factors affect the recovery of porcine fecal calprotectin,and therefore the researcher should double check certain parameters in regard to the type of kit,the dilution factor and homogenization time if reliable and reproducible results are to beobtained. Results of the present study provide useful information on a non-invasive protocol toveterinarians and researchers in examining and monitoring swine gut healthusing the fecalcalprotectin.
참고문헌 (25)
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