Analytical validation of an enzyme‐linked immunosorbent assay for the quantification of S100A12 in the serum and feces of cats

Bridges, Cory Seth; Grützner, Niels; Suchodolski, Jan S.; Steiner, Jörg M.; Heilmann, Romy M.

doi:10.1111/vcp.12812

Cited by 4 publications

(8 citation statements)

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“… 34 , 53 – 55 A canine S100A12-ELISA was first developed and shown to be accurate, reproducible, and sufficiently linear and precise for the measurement of S100A12 in cat serum and fecal samples. 55 A reference interval (RI) for feline fecal S100A12 was reported in a previous study. 55 However, since this RI has not been updated and the assay is only used infrequently, a statistical comparison between groups was performed rather than comparing results to the previously reported RI.…”

Section: Methodsmentioning

confidence: 99%

“… 55 A reference interval (RI) for feline fecal S100A12 was reported in a previous study. 55 However, since this RI has not been updated and the assay is only used infrequently, a statistical comparison between groups was performed rather than comparing results to the previously reported RI.…”

Section: Methodsmentioning

confidence: 99%

“…8,52 Fecal S100A12 concentrations were measured at the Gastrointestinal Laboratory at TAMU using an analytically validated in-house ELISA. 34,[53][54][55] A canine S100A12-ELISA was first developed and shown to be accurate, reproducible, and sufficiently linear and precise for the measurement of S100A12 in cat serum and fecal samples. 55 A reference interval (RI) for feline fecal S100A12 was reported in a previous study.…”

Section: Sample Analysesmentioning

confidence: 99%

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Fecal S100A12 concentrations in cats with chronic enteropathies

Zornow

Slovak

Lidbury

et al. 2023

Journal of Feline Medicine and Surgery

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Objectives The aim of this study was to compare fecal S100A12 concentrations in cats diagnosed with chronic enteropathy (CE) with healthy control cats. Methods This was a prospective, cross-sectional study. Forty-nine cats that had gastrointestinal signs for >3 weeks and a complete diagnostic work-up, including bloodwork, abdominal ultrasound and upper and/or lower gastrointestinal endoscopic biopsies, were enrolled into the CE group. Nineteen cats from the CE group were diagnosed with inflammatory bowel disease (IBD) or chronic inflammatory enteropathy (CIE) and 30 with alimentary lymphoma (LSA), based on histopathology results and additional testing with immunohistochemistry or molecular clonality testing with PCR if indicated. Nineteen apparently healthy control cats were included in the study. One fecal sample was collected from each cat and S100A12 concentrations were quantified by an analytically validated in-house ELISA. Results Fecal S100A12 concentrations differed between cats with LSA (median 110 ng/g; interquartile range [IQR] 18-548) and control cats (median 4 ng/g; IQR 2-25 [ P <0.001]) and between cats with IBD (median 34 ng/g; IQR 15–973) and control cats ( P <0.003). S100A12 concentrations in CE cats (median 94 ng/g; IQR 16–548) were statistically significantly higher compared with control cats ( P <0.001). The area under the receiver operating characteristic curve (AUROC) to separate healthy cats from CE cats was 0.81 (95% confidence interval [CI] 0.70–0.92) and was statistically significant ( P <0.001). The AUROC to separate cats with IBD from cats with LSA was 0.51 (95% CI 0.34–0.68) and was not statistically significant ( P = 0.9). Conclusions and relevance Fecal S100A12 concentrations at the time of diagnostic investigation were higher in cats with CIE and LSA than in healthy controls but did not differ between cats with LSA and those with CIE/IBD. This study is an initial step toward evaluating a novel non-invasive marker of feline CIE. Further studies are needed to determine the diagnostic utility of fecal S100A12 concentrations in cats with CE, including comparing cats with IBD/CIE and LSA, and to compare them with cats with extra-gastrointestinal disease.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Sample Analysesmentioning

confidence: 99%

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Fecal S100A12 concentrations in cats with chronic enteropathies

Zornow

Slovak

Lidbury

et al. 2023

Journal of Feline Medicine and Surgery

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“…Polyclonal rabbit antibodies generated against canine S100A8/A9 [ 49 , 50 ] and canine S100A12 protein [ 51 , 52 , 53 ] were used for IHC analysis of S100A8/A9 and S100A12 expression. Cross-reactivity of these polyclonal antibodies with feline S100A8/A9 and feline S100A12 has been demonstrated in immunodiffusion experiments and xenospecies immunoassays employing these antibodies or antiserum [ 49 , 51 , 54 , 55 ]. Surplus materials provided by the autopsy service at the Institute of Veterinary Pathology at UL-CVM were used as IHC controls.…”

Section: Methodsmentioning

confidence: 99%

Intestinal S100/Calgranulin Expression in Cats with Chronic Inflammatory Enteropathy and Intestinal Lymphoma

Riggers

Gurtner

Protschka

et al. 2022

Animals

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Diagnosing chronic inflammatory enteropathies (CIE) in cats and differentiation from intestinal lymphoma (IL) using currently available diagnostics is challenging. Intestinally expressed S100/calgranulins, measured in fecal samples, appear to be useful non-invasive biomarkers for canine CIE but have not been evaluated in cats. We hypothesized S100/calgranulins to play a role in the pathogenesis of feline chronic enteropathies (FCE) and to correlate with clinical and/or histologic disease severity. This retrospective case-control study included patient data and gastrointestinal (GI) tissues from 16 cats with CIE, 8 cats with IL, and 16 controls with no clinical signs of GI disease. GI tissue biopsies were immunohistochemically stained using polyclonal α-S100A8/A9 and α-S100A12 antibodies. S100A8/A9+ and S100A12+ cells were detected in all GI segments, with few significant differences between CIE, IL, and controls and no difference between diseased groups. Segmental inflammatory lesions were moderately to strongly correlated with increased S100/calgranulin-positive cell counts. Clinical disease severity correlated with S100A12+ cell counts in cats with IL (ρ = 0.69, p = 0.042) and more severe diarrhea with colonic lamina propria S100A12+ cells with CIE (ρ = 0.78, p = 0.021) and duodenal S100A8/A9+ cells with IL (ρ = 0.71, p = 0.032). These findings suggest a role of the S100/calgranulins in the pathogenesis of the spectrum of FCE, including CIE and IL.

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“…Although development and validation of assays for the measurement of certain APPs in cats' feces have been reported, [17][18][19] to the authors' knowledge, only α1-proteinase inhibitor (α1-PI) has been evaluated in cats with CE. 20 Our hypothesis was that certain APPs could be useful as a biomarker to diagnose, differentiate, and monitor cats with CE.…”

Section: Introductionmentioning

confidence: 99%

Fecal acute phase proteins in cats with chronic enteropathies

Karra,

Chadwick,

Stavroulaki

et al. 2023

Veterinary Internal Medicne

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BackgroundChronic enteropathies (CE) are common in cats and reliable biomarkers that can distinguish different causes and predict or monitor response to treatment are currently lacking.HypothesisTo evaluate certain acute phase proteins in feces that could potentially be used as biomarkers in cats with CE.AnimalsTwenty‐eight cats with either inflammatory bowel disease (IBD; n = 13), food‐responsive enteropathy (FRE; n = 3) or small cell gastrointestinal lymphoma (SCGL; n = 12) and 29 healthy control cats were prospectively enrolled.MethodsFecal concentrations of haptoglobin, alpha‐1‐acid‐glycoprotein (AGP), pancreatitis‐associated protein‐1 (PAP‐1), ceruloplasmin, and C‐reactive protein (CRP) were measured using Spatial Proximity Analyte Reagent Capture Luminescence (SPARCL) immunoassays before and after initiation of treatment. Cats were treated with diet and/or prednisolone (IBD cats), plus chlorambucil (SCGL cats).ResultsCompared with controls, median fecal AGP concentrations were significantly lower (25.1 vs 1.8 μg/g; P = .003) and median fecal haptoglobin (0.17 vs 0.5 μg/g), PAP‐1 (0.04 vs 0.4 μg/g) and ceruloplasmin (0.15 vs 4.2 μg/g) concentrations were significantly higher (P < .001) in cats with CE. Median fecal AGP concentrations were significantly lower (P = .01) in cats with IBD and FRE (0.6 μg/g) compared with cats with SCGL (10.75 μg/g). A significant reduction was found in CE cats after treatment for median fecal ceruloplasmin concentrations (6.36 vs 1.16 μg/g; P = .04).ConclusionsFecal AGP concentration shows promise to differentiate cats with SCGL from cats with IBD and FRE. Fecal ceruloplasmin concentrations may be useful to objectively monitor response to treatment in cats with CE.

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Analytical validation of an enzyme‐linked immunosorbent assay for the quantification of S100A12 in the serum and feces of cats

Cited by 4 publications

References 15 publications

Fecal S100A12 concentrations in cats with chronic enteropathies

Fecal S100A12 concentrations in cats with chronic enteropathies

Intestinal S100/Calgranulin Expression in Cats with Chronic Inflammatory Enteropathy and Intestinal Lymphoma

Fecal acute phase proteins in cats with chronic enteropathies

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