Evaluation of Cystatin C for the Detection of Chronic Kidney Disease in Cats.

BACKGROUND: Serum cystatin C (sCysC) and urinary cystatin C (uCysC) are potential biomarkers for early detection of chronic kidney disease (CKD) in cats. An in-depth clinical validation is required. OBJECTIVES: To evaluate CysC as a marker for CKD in cats and to compare assay performance of the turbidimetric assay (PETIA) with the previously validated nephelometric assay (PENIA). ANIMALS: Ninety cats were included: 49 CKD and 41 healthy cats. METHODS: Serum CysC and uCysC concentrations were prospectively evaluated in cats with CKD and healthy cats. Based on plasma exo-iohexol clearance test (PexICT), sCysC was evaluated to distinguish normal, borderline, and low GFR. Sensitivity and specificity to detect PexICT < 1.7 mL/min/kg were calculated. Serum CysC results of PENIA and PETIA were correlated with GFR. Statistical analysis was performed using general linear modeling. RESULTS: Cats with CKD had significantly higher mean ± SD sCysC (1.4 ± 0.5 mg/L) (P < .001) and uCysC/urinary creatinine (uCr) (291 ± 411 mg/mol) (P < .001) compared to healthy cats (sCysC 1.0 ± 0.3 and uCysC/uCr 0.32 ± 0.97). UCysC was detected in 35/49 CKD cats. R(2) values between GFR and sCysC or sCr were 0.39 and 0.71, respectively (sCysC or sCr = µ + GFR + Îµ). Sensitivity and specificity were 22 and 100% for sCysC and 83 and 93% for sCr. Serum CysC could not distinguish healthy from CKD cats, nor normal from borderline or low GFR, in contrast with sCr. CONCLUSION: Serum CysC is not a reliable marker of reduced GFR in cats and uCysC could not be detected in all CKD cats.

The effect of feeding, storage and anticoagulant on feline serum cystatin C.

Serum cystatin C (sCysC) is a possible marker for early detection of chronic kidney disease (CKD) in cats. In contrast with serum creatinine (sCr), feline sCysC is not affected by age, breed or sex. However, further biological and clinical validation is required. The objectives of this study were: (1) to investigate if food intake and circadian rhythm affect feline sCysC; (2) to determine the stability of sCysC under different storage conditions, and (3) to investigate if plasma concentrations of CysC (pCysC) differed from sCysC. A crossover study with 10 healthy laboratory cats fed the same commercial dry food was performed to study the influence of feeding and diurnal variation. Storage effects and comparison of pCysC with sCysC were determined using healthy cats (n = 3 and n = 10, respectively) and cats with CKD (n= 4 and n = 17, respectively). A significant daily sCysC variation was seen. Pre- and postprandial sCysC and sCr concentrations did not change significantly. Serum CysC significantly increased during storage at room temperature. After freezing, sCysC significantly decreased after 5 and 12 months at both -20 °C and -72 °C. Plasma CysC was significantly lower than sCysC. These findings suggest that it is not mandatory to fast cats before evaluation of sCysC and sCr. Samples were stable during routinely used storage conditions. Based on these findings, freezing for more than 5 months is not recommended, although additional studies are required to evaluate the clinical relevance of decreased sCysC after prolonged storage. Plasma and serum CysC cannot be compared directly.

Biological validation of feline serum cystatin C: The effect of breed, age and sex and establishment of a reference interval.

Chronic kidney disease (CKD) is common in cats, but the routine renal markers, serum creatinine (sCr) and urea, are not sensitive or specific enough to detect early CKD. Serum cystatin C (sCysC) has advantages over sCr, both in humans and dogs, and sCysC concentration is significantly higher in cats with CKD than in healthy cats. The objective of this study was to determine the effect of age, sex and breed on feline sCysC and to establish a reference interval for feline sCysC. In total, 130 healthy cats aged 1-16 years were included. sCysC was determined using a validated particle-enhanced nephelometric immunoassay. sCr, urea, urine specific gravity, urinary protein:creatinine ratio (UPC) and systolic blood pressure (SBP) were also measured. No significant differences in sCysC concentration were observed among young, middle-aged and geriatric cats, female intact, female neutered cats, male intact and male neutered cats, or among purebred and domestic short-or longhaired cats. The 95% reference interval for feline sCysC was determined to be 0.58-1.95 mg/L. sCr was significantly higher in geriatric cats than young cats. Serum urea in geriatric cats was significantly higher than in middle-aged and young cats (P = 0.004 and P <0.001, respectively). SBP in geriatric cats was significantly higher than in both middle-aged and young cats (P = 0.004 and P = 0.040, respectively). Male neutered and female neutered cats had significantly higher serum urea concentrations than female intact cats (P = 0.003 and P = 0.006, respectively). Male intact cats had a significantly higher UPC than female intact and female neutered cats (P = 0.02 for each comparison). There were no significant differences among sex groups for USG. It is of concern that sCysC in the majority of cats with CKD in previous studies falls within the reference interval calculated in this study. Further studies are warranted to evaluate the diagnostic value of sCysC as a renal marker in cats.