Comparison of Multistix PRO dipsticks with other biochemical assays for determining urine protein (UP), urine creatinine (UC) and UP:UC ratio in dogs and cats

BACKGROUND: Urine protein: urine creatinine (UP:UC) ratio determined from the quantitative measurement of protein and creatinine in a single urine sample is the best feasible assessment of clinically significant proteinuria in dogs and cats. A dipstick that measures urine protein, urine creatinine, and UP:UC ratio has been used in human medicine and could have application for veterinary practice. OBJECTIVE: The objective of this study was to compare the Multistix PRO dipstick (Bayer Corporation, Elkhart, IN, USA) to other biochemical methods for determination of urine protein and creatinine, and UP:UC ratio in canine and feline urine. METHODS: A complete urinalysis, including sulfosalicylic acid (SSA) precipitation, was performed on urine samples submitted to our laboratory between February and April 2003 from 100 dogs and 49 cats. Urine protein and creatinine concentrations were determined by the Multistix PRO dipstick using a Clinitek 50 analyzer (Bayer) and compared with the results of SSA precipitation and quantitative biochemical analysis. The UP:UC ratios from the dipstick results (calculated by the Clinitek 50 and also manually) were compared with those calculated from quantitative values. Pearson product-moment correlation analysis and diagnostic sensitivity and specificity (using quantitative results as the gold standard) were determined. RESULTS: For both canine and feline urine, protein and creatinine concentrations determined by the Multistix PRO correlated closely with quantitative concentrations for protein (dogs r = .78, P = .0001; cats r = .87, P = .0001) and creatinine (dogs r = .78, P = .0001; cats r = .76, P = .0001). The Multistix PRO was more sensitive and less specific than SSA precipitation for diagnosing clinically significant proteinuria. UP:UC ratios obtained by manual calculation of dipstick results correlated best with quantitative UP:UC ratios in dogs, and had higher specificity but lower sensitivity for the diagnosis of proteinuria. In cats, UP:UC ratios determined by the dipstick method did not correlate (r = -.24, P = .0974) with quantitative values. CONCLUSIONS: The Multistix PRO, with manual calculation of UP:UC, may be a good alternative for the diagnosis of clinically significant proteinuria in dogs, but not cats. Dipstick creatinine concentration should be considered as an estimate.     

Influence of urine pH on accurate urinary protein determination in Sprague-Dawley rats

BACKGROUND: Rat urinary protein concentration is commonly measured during safety assessment studies to evaluate potential drug-induced nephrotoxicity. It has been reported that impregnated reagent test strips (dipsticks) can yield false-positive urinary protein results for alkaline urine samples. OBJECTIVE: The objective of this study was to determine if urinary dipsticks accurately assess protein concentrations, especially in alkaline rat urine. METHODS: Ten male Sprague-Dawley rats were treated with 2% sodium bicarbonate and 2% ammonium chloride to alkalinize and acidify the urine, respectively. Urine pH was measured in treated and control rats using a pH meter and urinary dipsticks with the Clinitek 500. Quantitative urinary protein results were compared to urinary dipstick protein evaluations obtained with the Clinitek 500 and sulfosalicylic acid precipitation test methods. RESULTS: The urinary dipstick pH measurement had a very high correlation (r = .98) with the pH meter technique. Samples with alkaline pH (>or=7.5) analyzed for protein by dipstick analysis were in complete agreement 34.7% of the time with the quantitative technique, which was very similar to the 39.3% agreement for samples with neutral and acidic pH (相似文献   

The prevalence of albuminuria in dogs and cats in an ICU or recovering from anesthesia

Objective – To evaluate the prevalence of albuminuria in dogs and cats admitted to the ICU or recovering from an anesthetic event. Design – Prospective clinical study over a 10‐week period in 2003. Setting – Veterinary teaching hospital. Animals – One hundred and five dogs and 22 cats. Interventions – Urine was collected from dogs and cats admitted to the ICU or recovering from an anesthetic event. When possible, a second urine sample was collected approximately 48 hours later from those animals that had albuminuria during the initial screening. Measurements and Main Results – All dog samples and most cat samples were screened for albumin using a commercial point‐of‐care immunoassay. Aliquots of samples that tested positive were stored at –20°C until subsequent albumin quantification via antigen capture ELISA. Albuminuria was detected in 63 of 105 (60.0%) dogs and in 14 of 22 (63.6%) cats; the prevalence was higher in animals admitted to ICU than in those recovering from anesthesia. In subsequent samples from 26 dogs, urine albumin decreased in 20 (76.9%) when compared with the first sample; urine albumin was undetectable in 5 (19.2%). In subsequent samples from 6 cats, 4 (66.7%) had decreases in urine albumin when compared with the first sample; 1 (16.7%) was negative for urine albumin. Eleven of 12 dogs (91.7%) and 3 of 4 cats (75%) that died within 3 days of admission to the ICU had abnormal urine albumin; whereas 52 of 93 (55.9%) and 11 of 18 (61.1%) dogs and cats, respectively, who survived more than 3 days had abnormal urine albumin. Dogs with albuminuria were at increased risk of death. Conclusions – The prevalence of albuminuria in animals admitted to the ICU or recovering from anesthesia is higher than reported previously and transient in some patients. The presence of albuminuria may be a negative prognostic indicator in this population.     

Evaluation of IGF-I levels and serum protein profiles of diabetic cats and dogs

In this study, we measured the insulin-like growth factor (IGF)-I levels and evaluated the serum protein profiles of diabetic, insulin-treated, and healthy cats and dogs. The total IGF-I concentrations were 33.74 ± 3.4 ng/mL for normal, 25.8 ± 4.5 ng/mL for diabetic, and 180.4 ± 31.4 ng/mL for insulin-treated cats. IGF-I concentrations were 46.4 ± 6.6 ng/mL for normal, 25.1 ± 4.1 ng/mL for diabetic, and 303.0 ± 61.3 ng/mL for insulin-treated dogs. Total serum protein profiles were analyzed by SDS-PAGE. Fourteen bands ranging from 25 to 240 kDa in size were observed for cats, and 17 bands ranging from 25 to 289 kDa were observed for dogs. The densities of the bands differed among control, diabetic, and insulin-treated animals. In conclusion, we found that serum protein profiles and IGF-I concentrations were altered in both diabetic and insulin-treated animals. When judiciously interpreted in the light of other clinical and laboratory data, the techniques used in our study provide a valuable modality for measuring the severity of diabetes mellitus in dogs and cats.     

Effects of dietary crude protein and tannic acid on nitrogen excretion,urinary nitrogenous composition and urine nitrous oxide emissions in beef cattle

Two consecutive trials were carried out to study the effects of dietary crude protein (CP) and tannic acid (TA) on nitrogen (N) metabolism of beef cattle and consequently, the N₂O emissions from the urine of cattle. In Trial I, eight growing castrated cattle were used as the experimental animals. Two levels of dietary CP (110.6 and 135.7 g/kg dry matter [DM]) and two levels of TA (0 and 16.9 g/kg DM) were allocated in a replicated 2 × 2 crossover design. In Trial II, the N₂O emissions from the urine of cattle collected from Trial I were determined using the static incubation technique. An interaction between dietary CP and TA on the urinary N excretion (p < .05) was found but not on the N₂O‐N emission of cattle urine. Increasing dietary CP level from 110.6 g/kg DM to 135.7 g/kg DM increased the total N excretion (p < .001), the N retention (p < .05) and the ratio of urinary urea‐N/urinary N (p < .01), did not affect the N use efficiency (NUE; p > .05) and shifted the N excretion from faeces to urine. Increasing the dietary CP level increased the N₂O‐N emission of cattle urine. Dietary addition of TA decreased the urinary excretions of urea (p < .001) and shifted the N excretion from urine to faeces, did not affect the NUE of beef cattle (p > .10), and decreased the N₂O‐N emission of cattle urine. Pyrogallol and resorcinol of the TA metabolites were detected in urine with dietary addition of TA. Feeding beef cattle with relatively low CP level and adding TA in rations are effective approaches to mitigate the N₂O‐N emissions from cattle urine.     

Screening for proteinuria in cats using a conventional dipstick test after removal of cauxin from urine with a Lens culinaris agglutinin lectin tip

Proteinuria is an important indicator of urinary tract disease and urine dipsticks are simple and sensitive tools to screen for this marker. However, the use of dipsticks to screen for proteinuria may not be appropriate in cats, since cauxin, a 70 kDa glycoprotein, is secreted by the kidneys in clinically normal animals of this species. To circumvent this problem, a Lens culinaris agglutinin (LCA) lectin tip was developed to remove cauxin from feline urine, followed by conventional urine dipstick testing for proteinuria. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) with Coomassie brilliant blue R-250 staining indicated that >90% cauxin in the urine of 13 clinically normal cats was trapped by the LCA lectin tip, so that the dipstick protein ‘score’ changed from ‘positive’ (?30 mg/dL) for untreated urine to ‘negative’ (?10 mg/dL) for lectin tip-treated urine. In contrast, SDS–PAGE indicated that lectin tip-treated samples from 20 animals with renal disease contained high concentrations of albumin and low-molecular weight proteins; dipstick testing of lectin tip-treated urine resulted in a consistently positive protein score. The accuracy of the dipstick method for detecting cats with abnormal proteinuria is enhanced if dipsticks are used with urine samples that have first been passed through the LCA lectin tip.     

Acute phase proteins in dogs and cats: current knowledge and future perspectives

The acute phase response is a nonspecific inflammatory reaction of the host that occurs shortly after any tissue injury. The response includes changes in the concentration of plasma proteins called acute phase proteins (APPs), some of which decrease in concentration (negative APPs), such as albumin or transferrin, and others of which increase in concentration (positive APPs), such as C-reactive protein, serum amyloid A, haptoglobin, alpha-1-acid glycoprotein, and ceruloplasmin. Most positive APPs are glycoproteins synthesized mainly by hepatocytes upon stimulation by proinflammatory cytokines and released into the bloodstream. The acute phase response and clinical application of monitoring APPs in dogs and cats are reviewed in this article, including biochemical characteristics, assays developed for each individual APP, and preanalytic and analytic factors influencing APP results that should be taken into account for proper and adequate clinical interpretation. In addition, the diagnostic use of APPs and their possible application in monitoring treatment, which can be considered one of the most interesting and promising practical applications of these proteins, will be discussed. Finally, challenges and future developments of APPs in dogs and cats will be considered, because it is expected that new and cheaper automated assays for determination of the main APPs in small animals will contribute to a wider use of these proteins as biomarkers of infection and inflammatory lesions.     

Effects of crude protein and sodium intake on water turnover in cats fed extruded diets

The comprehension of strategies to increase urine production may be important, especially in kibble diets to prevent urolithiasis in cats. The effects of increasing amounts of crude protein (CP) and sodium on the water turnover of cats were evaluated using the water balance (WB) method and the deuterium dilution technique. The study followed a randomized block design, with three blocks of eight cats, two cats per food type in each block, and six cats per food. Four extruded diets with different amounts of CP and sodium were evaluated (on DM basis): 28% CP and 0.58% sodium; 39% CP and 0.64% sodium; 52% CP and 0.76% sodium; and 64% CP and 0.87% sodium. Cats were individually housed in cages for 8 days to measure WB, urea excretion, and faecal and urine characteristics. Deuterium oxide was used to evaluate water turnover, and during the period cats were housed in a collective cattery. The data were analysed by an F test, and the means were compared by polynomial contrasts. The ɑ level of significance was set at 0.05. The methods were compared by Pearson correlation, and Bland and Altman analysis. The increase in the CP content elevated linearly the renal excretion of urea (p < .001), and, together with the higher sodium intake, elevated the renal solute load, which resulted in a linear increase in urine production and water intake (p < .01). The urine density, metabolic water, and faecal and insensible water losses did not differ (p > .05). The water flux increased linearly when using the deuterium method (p < .001), but the obtained values were 20.85 ± 11.11 ml/cat/day higher than those verified using the WB method (p = .001). Higher CP and sodium amounts in dry diets increased the urine production and water consumption of cats, and this can be explored as a possible option to increase urination.     

Laboratory procedures for the diagnosis of gastrointestinal tract diseases of dogs and cats

An increasing number of laboratory tests are available for diagnosis of gastrointestinal tract diseases in dogs and cats. Use of these tests can lead to more accurate and rapid diagnoses. This review discusses laboratory tests, both new and old, and the role they currently play in the evaluation of animals presented with gastrointestinal problems. A minimum database helps assess the severity of the disorder, detect extra-gastrointestinal causes of problems and assists in formulating diagnostic and therapeutic plans.

Faecal examination remains one of the most important diagnostic procedures in the investigation of gastrointestinal problems. Zinc sulphate faecal flotation is an excellent routine screening technique for helminth and protozoal infections, including giardiasis. Rectal cytology can assist in the diagnosis of large bowel disorders. Interpretation of faecal immunodiagnostic tests is hampered by insufficient knowledge of test sensitivities and specificities. Routine faecal cultures are not warranted and faecal occult blood tests are rarely indicated.

Serum tests for gastric inflammation are now under development. The serum trypsin-like immunoreactivity test remains the gold standard technique for the diagnosis of exocrine pancreatic insufficiency. Breath hydrogen tests can be helpful in assessing the functional relevance of mild abnormalities in small-bowel biopsy specimens. Subnormal concentrations of serum cobalamin appear to be more specific indicators of gastrointestinal disease in cats than in dogs. Tests for small intestinal bacterial overgrowth remain controversial and assessment of gastrointestinal permeability has yet to prove its value in the diagnostic assessment of companion animals with gastrointestinal problems. Faecal alpha₁-protease inhibitor (α₁-PI) shows promise for the diagnosis of protein-losing enteropathy.     

Effect of storage time on the urine protein: creatinine ratio in alkaline ovine urine

The urine protein:creatinine (UPC) ratio is considered the reference method to assess proteinuria. Its diagnostic value in ovine medicine needs further elucidation. In population monitoring and/or for research purposes, it is convenient to collect many samples simultaneously and store them for later analysis. However, analyte stability data are required to ensure reliable results. We used 15 of 90 urine samples collected from sheep to assess the effect of storage time on the UPC ratio. After centrifugation, the supernatant of each sample was divided into 6 aliquots. Urine protein and creatinine concentrations were determined immediately in one aliquot using the pyrogallol red and a modified Jaffè method, respectively. The other aliquots were stored at −18°C. Based on the absence of active sediment, alkaline urine pH, and UPC ratio ≥0.2, we included 15 samples in our study. The UPC ratio was determined in the stored aliquots 2, 7, 14, 21, and 60 d after collection. The data were analyzed with univariate ANOVA. No significant difference was observed in the urinary concentrations of protein, creatinine, and the UPC ratio (0.8 ± 0.84 in conventional units and 0.09 ± 0.095 in SI units) among different times (p > 0.05). The UPC ratio remained stable for 2 mo in ovine urine samples stored at −18°C.     

Evaluating the use of plasma hematocrit samples to detect ketones utilizing urine dipstick colorimetric methodology in diabetic dogs and cats

Objective: To determine whether plasma from a heparinized hematocrit tube placed on a urine dipstick would accurately reflect (positive or negative) urine ketone results in diabetic dogs and cats. Design: Prospective study, 37 dogs and 43 cats, with a known history of diabetes or hyperglycemia, glucosuria, and symptoms of undiagnosed diabetes mellitus were tested. Setting: Veterinary Referral Hospital. Animals: Client owned dogs and cats. Interventions: None. Measurement and main results: Heparinized plasma and urine ketone results were recorded using urine reagent strips. Plasma dipstick results were compared to urine dipstick results as the standard. Results were recorded based on the color chart provided by the manufacturer. Two individuals were responsible for verifying the results of the colorimetric test. Test efficiency was 97% (sensitivity = 96%, specificity = 100%) for the canine population, 93% (sensitivity = 100%, specificity = 83%) for the feline population, and 95% (sensitivity = 98%, specificity = 91%) for the total population. Four of 80 animals were found to have discordant results (1 dog and 3 cats). Conclusion: Plasma from heparinized hematocrit tubes is clinically useful for detecting the presence or absence of ketonuria, and therefore ketosis, in diabetic dogs and cats using urine dipstick colorimetric methodology.     

C-reactive protein as an indicator of inflammatory responses to experimentally induced cystitis in dogs

The aim of this study was to demonstrate and assess C-reactive protein (CRP) changes in dogs with induced bacterial cystitis with or without antibiotics. We also evaluated availability of CRP levels to serve as an indicator for monitoring or diagnosing bacterial cystitis. Serial CRP concentrations in dogs with induced bacterial cystitis were higher than those of controls (p < 0.001). CRP concentrations peaked on day 7 and gradually decreased thereafter. In the treatment group, CRP concentrations decreased after medication compared to the untreated group (p = 0.032). CRP levels had a linear correlation with urine white blood cell counts among all groups (r = 0.837, p < 0.001, n = 140). Compared to the negative urine culture group, dogs with positive urine culture results had higher CRP concentrations (median 43.8 mg/L vs. 5.9 mg/L; p < 0.001). Area under the receiver operating characteristic curve was 0.955; when cut-off value was 12.2 mg/L, CRP measurements were found to have a sensitivity of 92.3% and specificity of 86.4%. This result indicates that rapid increases of CRP occurred after inducing bacterial cystitis and CRP may be a useful indicator for monitoring or diagnosing canine bacterial cystitis together with sediment urinalysis and urine bacterial culture.