TY - JOUR

T1 - Development of blood-based biomarker tests for early detection of colorectal neoplasia

T2 - Influence of blood collection timing and handling procedures

AU - Lech Pedersen, Niels

AU - Mertz Petersen, Mathias

AU - Ladd, Jon J.

AU - Lampe, Paul D.

AU - Bresalier, Robert S.

AU - Davis, Gerard J.

AU - Demuth, Christina

AU - Jensen, Sarah

AU - Andersen, Claus L.

AU - Ferm, Linnea

AU - Christensen, Ib J.

AU - Nielsen, Hans J.

PY - 2020/8

Y1 - 2020/8

N2 - Introduction: Blood-based, cancer-associated biomarkers are susceptible to a variety of well-known preanalytical factors. The influence of bowel preparation before a diagnostic colonoscopy on biomarker levels is, however, poorly investigated. The present study assessed the influence of bowel preparation on colorectal cancer-associated biomarkers. In addition, the effect of single versus double centrifugation of plasma biomarkers was assessed. Methods: Blood samples were collected pre- and post-bowel preparation from 125 subjects scheduled for first time diagnostic colonoscopy due to symptoms attributable to CRC. The samples were separated into serum and EDTA plasma, and analyzed by four independent collaborators for: 1) the proteins AFP, CA19-9, CEA, hs-CRP, CyFra21-1, Ferritin, Galectin-3 and TIMP-1, 2) the proteins BAG4, IL6ST, vWF, CD44 and EGFR, 3) the glycoprotein Galectin-3 ligand, and 4) cell-free DNA (cfDNA). Statistical analysis of biomarker data has been performed using mixed modelling, including repeated measures. Results: The biomarkers generally showed negligible variation between pre- and post-bowel preparation except for CyFra21-1, Ferritin, BAG4 and cfDNA. CyFra21-1 levels were systematically reduced with 29% (95% CI 21–36%) by bowel preparation (p ≤ 0.0001). Ferritin was not significantly different between pre- and post-bowel preparation (p = 0.07), however the estimated difference (increase) was 18%. BAG4 was systematically reduced by 12% (95% CI 1–22%, p = 0.04), while cfDNA showed a significant increase of 28% (95% CI 17–39%, p < 0.0001). Double centrifugation compared to single centrifugation showed reduced vWF (ratio 0.86, p ≤ 0.0001) and CD44 (ratio 0.85, p = 0.016), but increased IL6ST levels (ratio 1.18, p = 0.014). Conclusions: Results of the present study demonstrated systematic, statistically significant differences between pre-bowel and post-bowel preparation levels for three independent blood-based biomarkers (BAG4, CyFra21-1, cfDNA), illustrating the importance of timing of sample collection for biomarker analyses.

AB - Introduction: Blood-based, cancer-associated biomarkers are susceptible to a variety of well-known preanalytical factors. The influence of bowel preparation before a diagnostic colonoscopy on biomarker levels is, however, poorly investigated. The present study assessed the influence of bowel preparation on colorectal cancer-associated biomarkers. In addition, the effect of single versus double centrifugation of plasma biomarkers was assessed. Methods: Blood samples were collected pre- and post-bowel preparation from 125 subjects scheduled for first time diagnostic colonoscopy due to symptoms attributable to CRC. The samples were separated into serum and EDTA plasma, and analyzed by four independent collaborators for: 1) the proteins AFP, CA19-9, CEA, hs-CRP, CyFra21-1, Ferritin, Galectin-3 and TIMP-1, 2) the proteins BAG4, IL6ST, vWF, CD44 and EGFR, 3) the glycoprotein Galectin-3 ligand, and 4) cell-free DNA (cfDNA). Statistical analysis of biomarker data has been performed using mixed modelling, including repeated measures. Results: The biomarkers generally showed negligible variation between pre- and post-bowel preparation except for CyFra21-1, Ferritin, BAG4 and cfDNA. CyFra21-1 levels were systematically reduced with 29% (95% CI 21–36%) by bowel preparation (p ≤ 0.0001). Ferritin was not significantly different between pre- and post-bowel preparation (p = 0.07), however the estimated difference (increase) was 18%. BAG4 was systematically reduced by 12% (95% CI 1–22%, p = 0.04), while cfDNA showed a significant increase of 28% (95% CI 17–39%, p < 0.0001). Double centrifugation compared to single centrifugation showed reduced vWF (ratio 0.86, p ≤ 0.0001) and CD44 (ratio 0.85, p = 0.016), but increased IL6ST levels (ratio 1.18, p = 0.014). Conclusions: Results of the present study demonstrated systematic, statistically significant differences between pre-bowel and post-bowel preparation levels for three independent blood-based biomarkers (BAG4, CyFra21-1, cfDNA), illustrating the importance of timing of sample collection for biomarker analyses.

KW - AFP

KW - BAG4

KW - Biomarkers

KW - Bowel preparation

KW - CA19-9

KW - CD44

KW - CEA

KW - Centrifugation

KW - cfDNA

KW - Colorectal cancer

KW - CyFra21-1

KW - EGFR

KW - Ferritin

KW - Galectin-3

KW - Galectin-3 ligand

KW - hs-CRP

KW - IL6ST

KW - Preanalytical variation

KW - TIMP-1

KW - vWF

U2 - 10.1016/j.cca.2020.03.035

DO - 10.1016/j.cca.2020.03.035

M3 - Journal article

C2 - 32272156

AN - SCOPUS:85083238100

VL - 507

SP - 39

EP - 53

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

SN - 0009-8981

ER -