AI Article Synopsis
- The study aimed to develop a urine test for detecting kidney injury molecule 1 (KIM-1) in both healthy and diseased cats.
- Methods included creating and purifying proteins from engineered cells, immunizing mice, and constructing a lateral flow assay (LFA) tested with various cat urine samples.
- Results showed that the LFA could detect KIM-1 in feline urine, with higher levels found in cats with acute kidney injury (AKI), indicating its potential as a useful diagnostic tool for feline renal health.
Article Abstract
Objectives: The aim of this study was to design and carry out a preliminary evaluation of a urine point-of-care test for kidney injury molecule 1 (KIM-1) in healthy and diseased cats.
Methods: Part of the feline gene was amplified, ligated into a plasmid with a signal peptide and monomeric human IgGFc, and transfected into a mammalian cell line. Supernatant was purified and tested for the fusion protein by gel electrophoresis and Western blot. Mice were immunized three times with purified proteins, and hybridomas were generated from splenocytes. Antibodies were tested by ELISA for detection of recombinant KIM-1 and naturally occurring KIM-1 in disease-state urine. Next, a lateral flow assay (LFA) with capture and detection antibodies was constructed, and tested with 34 urine samples from healthy and diseased cats. Antibodies were also tested for reactivity with formalin-fixed paraffin-embedded kidney tissue.
Results: Three antibodies were assessed. Antibodies detected between 0.4 and 60 ng/ml feline KIM-1 fusion protein in the LFA. Urine samples from healthy cats yielded faint bands in the LFA corresponding to optical density (OD) values of 4.8-8.8. Samples from cats with suspected or confirmed acute kidney injury (AKI) had OD values ranging from 1.6-20.5. Urine KIM-1 varied over multiple days in cats with sepsis or urethral obstruction despite normalizing serum creatinine concentration. In tissue sections, KIM-1 antibodies labeled tubular cells with morphological features of injury.
Conclusions And Relevance: A practical patient-side assay for detection of KIM-1 in feline urine has been developed. Preliminary results show marked though transient increases in cats with sepsis and urethral obstruction-associated AKI, and expression in injured tubules. Although initial data indicating that the LFA is sensitive and specific for KIM-1 in cats with AKI are promising, values associated with different types of injury, urine collection, urine storage and specific gravity need to be investigated.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10814266 | PMC |
| http://dx.doi.org/10.1177/1098612X18812494 | DOI Listing |
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