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Науковий вгсникЛНУВМБТ 1мен1 С.З. Гжицького Том 10№ 2 (37) Частина 2, 2008
Pawe! Jonkisz, Agnieszka Sikorska, Agnieszka Kurosad, Igor Maksymovych*©
Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life
Sciences, Poland
*Lviv National University of Veterinary Medicine and Biotechnologies named after
S.Z. Gzhytskyj
COMPARISON OF CYSTATINE C LEVELS MEASURED IN DOGS BY TWO DIFFERENT METHODS: PENIA AND PETIA
Abstract
Cystatine C is a low-molecule protein which is suggested as a marker of chronic renal insufficiency in dogs. It is measured by radioimmunological methods, using rabbit antibodies against the human cystatine C and turbidimetric (PETIA) and nephelometic (PENIA) immunoassay.
40 dogs of various breeds, sex and age were divided into 4 groups, consist of 10 and cystatine C was measured in those groups using PENIA and PETIA, comparably. The urea and creatinine were also evaluated in dogs blood serum. The correlation coefficient for Cystatine C, measured by PENIA and PETIA, was 0,7055. Those coefficient between Cysatine C in PENIA and PETIA and creatinine was evaluated on: 0,90353 and 0,61428, respectively. The correlation coefficient of urea and cystatine C, measured by PENIA and PETIA were: 0,63591 and 0,46311, respectively.
Key words: renal insufficiency, cystatine C, PETIA, PENIA, dog.
Introduction
Cystatine C is a low-molecule protein, protease inhibitor, which is produced by most nucleus cells. Its synthesis do not change according to sex, age and muscle masses. Cystatine C is stabile according to inflammatory condition and easily filtrate by renal glomerule [7]. From 1985, in human medicine it is suggested as a marker of renal function. Although cystatine C is a good indicator for monitoring renal disease, some scientists prefer creatinine for it [4]. Laterza and Newman observed that cystatine C was increased earlier than creatinine level in dogs with renal insufficiency [12,14]. Up till now cystatine C is not suggested as a good marker of renal insufficiency in cats [13]. Cystatyne C in dogs can be measured by Western-blott method and subsequently by PETIA (Particle-enhanced turbidimetric immunoassay), by which method the cystatine C was evaluated in group of 50 dogs [1]. The mean level of cystatine C, measured by PETIA, in group of healthy dogs was: 1,08 ±0,16 mg/L, and in animals with renal insufficiency it was: 4,37 ±1,79mg/L. Nearly the same results were achieved by Jensen et al. and there were: 1,06 and 5,01, respectively [10]. However Gonul observed the greater difference among testing groups and the values of cystatine C were: 1,2 ±0,42 and 2,96 ±1,09, respectively [8].
© Pawel Jonkisz, Agnieszka Sikorska, Agnieszka Kurosad, Igor Maksymovych, 2008
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Hayxoeuu eicnux flHYBMET iMeni C.3. í^ицbкого
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PENIA (Particle-enhanced nephelometric immunoassay), which was described in details in 1997, is more specific method for measuring the cystatine C level than PETIA [7] and today it is usually used in most hospital laboratories. Nowadays, the lower reference range of cystatine C is proposed for healthy dogs and it is: 0,25±0,15 mg/L, measured by PETIA [3]. Cystatine C is rather stabile in room temperature and do not change during 10 cycles of freezing above -20 °C [7]. However, other scientists suggested various stability of cystatine C in various temperature for prolong term of time [6, 11,14].
The, the aim of the article
The main aim of the experiment was to compare two methods: PETIA and PENIA for measuring the level of cystatine C in dogs and establish the correlation coefficient among cystatine C, creatinine and urea in dogs.
Material and methods
40 dogs of various breeds, age and sex were divided into 4 groups, consisted of 10. The animals were classified into the proper group based on IRIS scale. Group no I consisted of healthy dogs and the creatinine level was: 125 mmol/l; group no II had the level of creatinine between: 125 and 180 mmol/l and the group no III and IV consisted of animals with chronic renal disease with the creatinine measured on the level between: 181 and 440 and above 440 mmol/l, respectively. Morphology and biochemical analyses were evaluated using the Animal Blood Counter model ABC Vet (ABX - France) and MAXMAT PL (Montpellier - France) apparatus. Cystatine C was measured by PETIA - Dako Cytomation Cystatin C Immunoparticles (Hitachi Modular) and PENIA - Dade Behring N Latex Cystatin C assay (Dade Behring Nephelometr II). Statistical analysis was done by the computer program Statistica 5 and the value of P<0,05 was taken as a statistically significant.
Results of researches All results were presented in tables no: 1-3:
"ab. 1. Comparison of cystatine C and urea mean levels.
"""""-Parrametr Cystatyna C/PENIA mg/l Cystatyna C/PETIA mg/l Mocznik mmol/l Kreatynina umol/l
Gr. I 0,207 ± 0,039 0,81 ± 0,166 5,21 ± 1,34 97,5 ± 10,7
Gr. II 0,326 ± 0,138 0,98 ± 0,162 6,30 ± 3,16 142,6 ± 13,5
Gr. III 0,593 ± 0,245 1,05 ± 0,217 30,19 ± 18,81 266,4 ± 63,1
Gr. IV 1,226 ± 0,646 1,18 ± 0,249 55,68 ± 16,92 780,5 ±256,3
The mean difference level of cystatine C, measured by PENIA and PETIA was: 0,417 ± 0,394
Gr. II Gr. III Gr. IV
Gr. I 0,0197 0,00077 0,00074
Gr. II 0,00952 0,00154
Gr. III 0,01342
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Науковий вгсник ЛНУВМБТ 1мен1 С.З. Гжицького
Том 10 № 2 (37) Частина 2, 2008
Tab. 3. Comparison of P for cystatine C (PETIA) between groups: II-IV
Gr. II Gr. III Gr. IV
Gr. I 0,03257 0,01301 0,00124
Gr. II 0,42534 0,04995
Gr. III 0,22903
Discussion
The experiment provided in 4 groups of dogs showed the significantly lower level of cystatine C, measured by PENIA than PETIA (tab.1). Similar experiment in human medicine showed a higher precision of PENIA for measuring the level of cystatine C than PETIA, greater difference between evaluated values (1,13 ± 0,86 mg/L) and good correlation between those two methods [7]. The value of cystatine C, measured by PENIA in group of healthy dogs was: 0,207 ± 0,039 and it was similar to the values of reference range proposed earlier. The value of cystatine C for groups of dogs with chronic renal failure was: 1,226 ± 0,646 and it was higher than estimated earlier [2,3]. Results of cystatine C obtained in PETIA for groups of healthy and dogs with chronic renal insufficiency were: 0,81 ± 0,166 and 1,18 ± 0,249, respectively and there were lower than measured by other scientists [1,9]. P values for cystatine C, estimated by PENIA were significantly lower than measured by PETIA. However, the statistically important differences for cystatine C levels, measured by PENIA were observed between all groups and by PETIA only among the group no II and IV. Those results suggest that PENIA is more precise method for evaluation the cystatine C level than PETIA and it should be used in clinical practice as a standard method [5]. However, in diagnosis of early renal insufficiency in dogs the value of cystatine C should be evaluated according to creatinine level and GRF [9] .
In statistical analysis the better correlation was observed between cystatine C, measured by PENIA and urea and creatinine level than the cystatine C evaluated by PETIA.
References:
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6. Erlandsen E.J., Randers E., Kristensen J.H.: Evaluation of Dade Behring N Latex Cystatin C assay on the Dade Behring Nephelometer II System. 1999,1,1-8,
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11. Kyhse-Andersen J., Schmidt C., Nordin G., Andersson B., Nilsson-Ehle P., Lindstrom V., Grubb A.: Serum cystatin C, determined by a rapid, automated particle-enhanced turbidimetric method, is a better marker than serum creatinine for glomerular filtration rate. Clin. Chem. 1994,40,1921-1926.
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Summary
Jonkisz P., Sikorska A., Kurosad A., Maksymovych I. * Department of Internal and Parasitic Diseases with Clinic for Horses, Dogs and Cats, Faculty of Veterinary Medicine, Wroclaw University of Environmental and
Life Sciences, Poland
*Lviv National University of Veterinary Medicine and Biotechnologies named after
S.Z. Gzhytskyj
COMPARISON OF CYSTATINE C LEVELS MEASURED IN DOGS BY TWO DIFFERENT METHODS: PENIA AND PETIA
The article describes the values of cystatine C, measured by two different
methods: PENIA and PETIA in 40 dogs divided in 4 groups, classified by IRIS scale.
The results of cystatine C measured by PENIA were more precise in all groups of
dogs.
Cmammx nadiümna do peda^ii 18.02.2008
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