The Long-term Biological Variability of Fasting Plasma Glucose and Serum Fructosamine in Healthy Beagle Dogs

The aim of this study was to estimate the long-term (month-to-month) between-dog, within-dog and analytical components of variance for fasting plasma glucose and serum fructosamine in healthy dogs to assess the usefulness of a single measurement of these analytes in a single dog.Fasting plasma glucose and serum fructosamine were measured in blood samples collected every month for 9 months from 23 clinically healthy dogs, and the results were subjected to nested analysis of variance. The between-dog variation, the within-dog variation, and the analytical variation were 3.8%, 9.5% and 3.7%, respectively, for plasma glucose and 4.2%, 11.1% and 2.8%, respectively, for serum fructosamine.The maximum allowable analytical imprecision, analytical inaccuracy and difference between analytical methods were 4.8%, 2.6% and 3.2%, respectively, for plasma glucose and 5.6%, 3.0% and 3.7%, respectively, for serum fructosamine.The index of individuality, 2.7 for both analytes, indicated that the test results from single dogs can be compared usefully to the corresponding population-based reference intervals.The number of samples required to estimate the true individual mean value ±5% for a single dog was 16 for fasting plasma glucose and 20 for serum fructosamine.The one- and two-sided critical differences expressing the difference needed for two serial results from the same dog to be significantly different at a 5% level was 24% and 28%, respectively, for plasma glucose and 27% and 32%, respectively, for serum fructosamine.     

Study on biological variation of haemostatic parameters in clinically healthy dogs

Thromboelastography (TEG) may be a valuable supplement to the coagulation assays activated partial thromboplastin time (aPTT), prothrombin time (PT), thrombin time (TT), fibrinogen, antithrombin (AT) and D-Dimer currently used in most clinical pathology laboratories. Allowable imprecision and bias reference limits for analytical tests can be calculated based on measurements of biological variation. No studies to date have examined the effect of biological variation on these haemostasis parameters in the same group of dogs. Plasma samples were collected after a set protocol once weekly for five consecutive weeks from eight healthy dogs (four males and four females) and stored at -80 degrees C until analysis. Randomized duplicate coagulation tests and TEG analyses were performed on all plasma samples within one run. The data were analyzed for outliers and subsequently subjected to nested analysis of variance to obtain the coefficient of analytical, intra-individual and inter-individual variation. From these objective analytical performance standards for imprecision, critical difference, total error and the index of individuality were calculated to assess the utility of conventional population-based reference ranges. All the clotting times (aPTT, PT and TT), fibrinogen, AT and D-Dimer showed a degree of individuality, which may make the use of population-based reference ranges alone an insensitive interpretation criterion, whereas a population-based reference interval seems to be sensitive for interpreting all TEG parameters. Analytical performance standards for imprecision were only met for one of the coagulation assays, whereas all TEG parameters except the alpha angle, alpha achieved this analytical goal.     

Biological Variability and Measurement Error Variability in Ocular Biometry in Samoyed Dogs

A-scan ultrasonography was performed in 40 healthy Samoyeds aged 2–5 years. Mydriasis and cycloplegia were induced in all dogs and 25 of the dogs were also sedated prior to the ultrasound examination. Five consecutive A-scans were taken on each eye and the intrasubject variance (measurement error) and true intersubject variance (true biological variation) were computed. The true biological variation was found to be of the same magnitude in both sedated and unsedated dogs, whereas the measurement errors were considerably larger in the group of unsedated dogs compared to the sedated ones. Magnitudes of the measurement errors for the anterior chamber depth and length of the vitreous body in the unsedated group were unacceptable. The sizes of the measurement errors in the sedated dogs were all within an acceptable range.     

Reference interval and critical difference for canine serum fructosamine concentration

The purposes of the study were to obtain a reference interval and to calculate the critical difference between two analytical results for canine serum fructosamine concentration. To obtain a reference interval, the serum fructosamine concentration was measured in blood samples from 29 adult dogs after a 15-h fasting period. To calculate the critical difference, blood samples from 20 apparently clinically healthy dogs were collected once weekly for five consecutive weeks, and the total variance of the analytical results was divided into the component of variance between dogs (S _inter ² ), the component of variance for weeks within dogs (S _intra ² ) and the component of variance for measurements (S _anal ² ), using nested analysis of variance. The critical difference was then calculated fromS _intra ² andS _anal ² .The main conclusions are in summary: The reference interval for canine serum fructosamine concentration is 258.6–343.8 µmol/L, and the critical difference between two consecutive measurements on a week-to-week basis is 32.4 µmol/L. The critical difference may be used as a guideline to indicate potentially important changes in the serum fructosamine concentration, though the analytical results should not be assessed by the critical differences alone, but should also be compared to the corresponding reference intervals.     

Effects of short-term trimethoprim-sulfamethoxazole administration on thyroid function in dogs

OBJECTIVE: To determine how rapidly trimethoprim-sulfamethoxazole affects serum total thyroxine (T4) and thyroid-stimulating hormone (TSH) concentrations in euthyroid dogs and how quickly hormone concentrations return to reference values following discontinuation of administration. DESIGN: Prospective study. ANIMALS: 7 healthy euthyroid dogs. PROCEDURE: Dogs were given trimethoprim-sulfamethoxazole (26.5 to 31.3 mg/kg [12 to 14.2 mg/lb], PO, q 12 h) for a maximum of 6 weeks. A CBC and Schirmer tear test were performed and serum total T4 and TSH concentrations were measured weekly. Administration of trimethoprim-sulfamethoxazole was discontinued if total T4 concentration was less than the lower reference limit and TSH concentration was greater than the upper reference limit or if persistent neutropenia developed. RESULTS: Six dogs had total T4 concentrations less than the lower reference limit within 3 weeks; T4 concentration was decreased after 1 week in 3 of these 6 dogs. In these 6 dogs, TSH concentration was greater than the upper reference limit within 4 weeks. In 1 dog, T4 and TSH concentrations were not affected, despite administration of trimethoprim-sulfamethoxazole for 6 weeks. Neutropenia developed in 4 dogs. In 1 dog, the neutropenia resolved while trimethoprim-sulfamethoxazole was still being administered. In the other 3, neutrophil counts returned to reference values 1 week after drug administration was discontinued. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of trimethoprim-sulfamethoxazole at a dosage of 26.5 to 31.3 mg/kg, PO, every 12 hours can substantially alter serum total T4 and TSH concentrations and neutrophil counts in dogs within as short a time as a few weeks.     

Serum concentrations of thyroxine and 3,5,3'-triiodothyronine in dogs before and after administration of freshly reconstituted or previously frozen thyrotropin-releasing hormone

Concentrations of serum thyroxine (T4) and 3,5,3'-triiodothyronine (T3) were determined after the administration of freshly reconstituted thyrotropin-releasing hormone (TRH), reconstituted TRH that had been previously frozen, or thyrotropin (TSH) to 10 mature dogs (6 Greyhounds and 4 mixed-breed dogs). Thyrotropin-releasing hormone (0.1 mg/kg) or TSH (5 U/dog) was administered IV; venous blood samples were collected before and 6 hours after administration of TRH or TSH. Concentrations of the T4 and T3 were similar (P greater than 0.05) in serum after administration of freshly reconstituted or previously frozen TRH, indicating that TRH can be frozen at -20 C for at least 1 week without a loss in potency. Concentrations of T4, but not T3, were higher after the administration of TSH than they were after the administration of TRH (P less than 0.01). Concentrations of T4 increased at least 3-fold in all 10 dogs given TSH, whereas a 3-fold increase occurred in 7 of 10 dogs given freshly reconstituted or previously frozen TRH. Concentrations of T4 did not double in 1 dog given freshly reconstituted TRH and in 1 dog given previously frozen TRH. Concentrations of T3 doubled in 5 of 10, 2 of 10, and 5 of 10 dogs given TSH, freshly reconstituted TRH, or previously frozen TRH, respectively. Results suggested that concentrations of serum T4 are higher 6 hours after the administration of TSH than after administration of TRH, using dosage regimens of 5 U of TSH/dog or 0.1 mg of TRH/kg.(ABSTRACT TRUNCATED AT 250 WORDS)     

Short-term biological variation of clinical chemical values in Dumeril's monitors (Varanus dumerili).

Plasma biochemical values are routinely used in the medical management of ill reptiles, and for monitoring the health of clinically normal animals. Laboratory tests, including clinical biochemical values, are subject to biological and analytical variation, the magnitude of which determines the utility of population-based reference ranges for the detection of abnormal results in the individual animal. Nested analysis of variance of repeated measurements allows the variance to be broken into within-individual, between-individual, and analytical variation. When the within-individual variation is large and the interindividual variation is low, a sample may be accurately classified as normal or abnormal based on a population-based reference interval. However, if the intraindividual variation is low and the interindividual variation high, population-based reference intervals are of limited value as the ranges for an individual encompass only a part of the conventional reference interval. Between-lizard, within-lizard, and analytical components of variance were assessed by nested analysis of variance for 16 commonly measured plasma biochemical parameters in eight healthy adult Dumeril's monitors (Varanus dumerili). Albumin, cholesterol, phosphate, calcium, sodium, and total protein demonstrated levels of individuality suggesting that comparison of a single measurement to a conventional population-based reference range may be too insensitive to detect small but significant alterations in the value for that animal. Only for potassium and AST did the index of individuality suggest that the use of reference values may be warranted. Uric acid, globulin, glucose, and amylase fell in a gray zone, where population-based ranges should be used with caution. The critical difference indicates the difference between two consecutive analytical results that may be safely ascribed to natural variation. In the present study critical difference varied from 7 and 11%, respectively, for sodium and chloride to 75 and 125% for uric acid and AST.     

Homocysteine measurement by an enzymatic method and potential role of homocysteine as a biomarker in dogs

In humans, homocysteine (Hcy) is employed to monitor renal, cardiovascular, and other diseases and their complications. The aim of the current study was to define the analytical performances of an enzymatic method not yet validated in dogs for measuring homocysteine, and to assess the possible clinical usefulness of Hcy measurement. Using conventional approaches, the analytical performances were investigated by assessing, imprecision, inaccuracy, and interference of hemoglobin, triglycerides, and bilirubin. The possible clinical usefulness of Hcy determination was assessed by comparing the results of healthy dogs (n = 8) with those of dogs with heart disease (n = 10), inflammation (n = 6), gastrointestinal disorders (n = 7), neoplasia (n = 8), renal failure (n = 4), trauma (n = 7), and other miscellaneous diseases (n = 6). Preliminary evaluation of this enzymatic method showed it to be precise at Hcy levels close to or higher than the values in dogs with renal or cardiac disorders that had the highest Hcy levels. By contrast, at low Hcy levels, which were recorded basically in control dogs, the method suffers from high imprecision. The sample of choice is serum. The use of icteric samples should be avoided, while hemoglobin and lipids have only a minor effect on Hcy measurement. In conclusion, the enzymatic method employed in the current study provides useful information in dogs and could be used to monitor cardiac and renal disorders, in which Hcy concentrations are elevated.     

Effect of Thyrotropin Storage on Thyroid-Stimulating Hormone Response Testing in Normal Dogs

The stability of reconstituted, refrigerated thyrotropin was evaluated. Thyrotropin (TSH) was reconstituted at the start of the study and stored at 4 degrees C. A TSH stimulation test was performed in eight healthy, euthyroid dogs at weekly intervals for 1 month. In seven of eight dogs, there was no significant difference (P less than 0.05) between the post-TSH T3 concentrations and the post-TSH T4 concentrations for the duration of the study. For one dog, the post-TSH T4 concentration was below the normal post-TSH T4 range following the administration of reconstituted TSH that had been stored 4 weeks. The T3 response to the TSH, however, was normal. This dog responded normally to freshly reconstituted TSH. The results of this study suggest that reconstituted bovine TSH can be stored at 4 degrees C for at least 3 weeks without loss of biologic activity in the dog.     

Evaluation of analytical performance assisted by total error criteria of a commercial enzyme immunometric assay for canine serum thyrotropin

The aim of the present study was to evaluate analytical performance using total error criteria of a commercial enzyme immunometric assay for the determination of endogenous canine thyrotropin (TSH). The allowable total error for this assay (22.6%) was estimated using previously reported data on biological variation. Inaccuracy and imprecision of the assay (0% and 5.7% for the low control material; 6.8% and 3.0% for the high control material) were estimated by measuring the same lot of control material for 21 consecutive weeks, during which time the assay was considered stable and in control. Analytical performance was assessed using a MEDx chart, a graphical tool for comparing inaccuracy and imprecision, with an analytical quality requirement stated in the form of allowable total error. The results of the present study showed that the canine TSH assay had good to excellent analytical performance.     

Effects of administration of a low dose of frozen thyrotropin on serum total thyroxine concentrations in clinically normal dogs.

Thyroid function was evaluated in 18 healthy dogs by thyrotropin (TSH) stimulation. Two dose regimens were used in each dog: 0.1 IU/kg body weight of freshly reconstituted lyophilized TSH and 1 IU/dog of previously frozen and stored TSH (up to 200 days), both given intravenously. Blood samples were collected prior to and at four and six hours after TSH administration. Serum was evaluated for total thyroxine concentrations by radioimmunoassay. All dogs were classified as euthyroid on the basis of response to 0.1 IU/kg body weight of freshly reconstituted TSH at four and six hours. The 1 IU dose of TSH, previously frozen for up to 200 days, induced increases in serum total thyroxine concentration over baseline at four and six hours that were not significantly different from those resulting from the use of the higher dose of fresh TSH. In all test groups, there were no statistically significant differences between total thyroxine concentrations at four and six hours post-TSH administration. It was concluded that an adequate TSH response can be achieved with the use of 1 IU of TSH/dog for clinically normal dogs between 29.0 kg and 41.6 kg body weight, even if this TSH has been frozen at -20 degrees C for up to 200 days. Further, blood collection can be performed at any time between four and six hours. Similar studies are needed to evaluate this new protocol in hypothyroid dogs and euthyroid dogs suffering nonthyroidal systemic diseases.     

Critical differences of acute phase proteins in canine serum samples

The critical difference values for acute phase proteins in canine serum samples were established on a week-to-week basis. Blood samples from 11 apparently clinically healthy dogs were collected once weekly for five consecutive weeks. For each protein the total variance of analytical results was divided into intraindividual variance (S(Intra)(2)), interindividual variance (S(Inter)(2)), and analytical variance (S(Analytical)(2)). The critical difference (d(K)) was then calculated as d(k)=22(S(Intra)(2)+S(Analytical)(2)). The critical difference values were 1.95 g/L for haptoglobin, 4.85 mg/L for C-reactive protein, and 0.016 DeltaAbs/min for ceruloplasmin. When used in conjunction with the corresponding reference interval, critical difference values can be an aid in correctly interpreting acute phase protein results, by determining whether observed differences between two consecutive measurements in individual animals are due to natural variation or due to disease therapy or experimental procedures.     

Renal perfusion parameters measured by contrast‐enhanced ultrasound in healthy dogs demonstrate a wide range of variability in the long‐term

Contrast‐enhanced ultrasound may be helpful for detecting early renal microvascular damage and dysfunction in dogs. However, before this noninvasive imaging method can be tested as an early‐stage screening tool in clinical patients, an improved understanding of long‐term variation in healthy animals is needed. In this prospective, secondary, longitudinal, serial measurements study, variability of contrast‐enhanced ultrasound renal perfusion parameters was described for eight healthy dogs, using seven time points and a period of 83 weeks. Dogs were sedated with butorphanol (0.4 mg/kg), and contrast‐enhanced ultrasound of each kidney was performed after an intravenous bolus injection of a microbubble contrast agent (0.04 mL/kg). Time‐intensity curves were created from regions‐of‐interest drawn in the renal cortex and medulla. Intensity‐related parameters representing blood volume and time‐related parameters representing blood velocity were determined. A random‐effects model using restricted maximum likelihood was used to estimate variance components. Within‐dog coefficient of variation was defined as the ratio of the standard deviation over the mean. Time‐related parameters such as time‐to‐peak, rise and fall time had lowest within‐dog variability. Intensity‐related parameters such as peak enhancement, wash‐in and wash‐out area under the curve, total area under the curve, and wash‐in and washout rates had high within‐dog variability (coefficient of variation > 45%). Authors therefore recommend the use of time‐related parameters for future studies of renal perfusion. Within‐dog variability for bilateral kidney measurements was extremely low, therefore contrast‐enhanced ultrasound may be particularly useful for detecting unilateral changes in renal perfusion. Future studies are needed to compare contrast‐enhanced ultrasound findings in healthy dogs versus dogs with renal disease.     

Evaluation of thyroid function in obese dogs and in dogs undergoing a weight loss protocol

Obesity and weight loss have been shown to alter thyroid hormone homeostasis in humans. In dogs, obesity is the most common nutritional problem encountered and weight loss is the cornerstone of its treatment. Therefore, it is important to clarify how obesity and weight loss can affect thyroid function test results in that species. The objectives of this study were to compare thyroid function in obese dogs and in lean dogs and to explore the effects of caloric restriction and weight loss on thyroid hormone serum concentrations in obese dogs. In the first experiment, 12 healthy lean beagles and 12 obese beagles were compared. Thyroid function was evaluated by measuring serum concentrations of total thyroxine (TT4), free thyroxine (FT4), total triiodothyronine (TT3), thyrotropin (TSH), and reverse triiodothyronine (rT3) as well as a TSH stimulation test using 75 microg i.v. of recombinant human TSH. In the second experiment, eight obese beagles were fed an energy-restricted diet [average 63% maintenance energy requirement (MER)] until optimal weight was obtained. Blood samples for determination of TT4, FT4, TT3, TSH and rT3, were taken at the start and then weekly during weight loss. Only TT3 and TT4 serum concentrations were significantly higher in obese dogs as compared to lean dogs. In the second experiment, weight loss resulted in a significant decrease in TT3 and TSH serum concentrations. Thus obesity and energy restriction significantly alter thyroid homeostasis in dogs, but the observed changes are unlikely to affect interpretation of thyroid function test results in clinics.     

Analytical and Clinical Validation of a Time-resolved Immunofluorometric Assay (TR-IFMA) for Canine C-reactive Protein in Serum

A time-resolved immunofluorometric assay (TR-IFMA) was developed for the determination of C-reactive protein (CRP) in canine serum. CRP was isolated from canine acute-phase serum by affinity chromatography on agarose coupled with phosphorylethanolamine. This isolated dog CRP was used as standard to calibrate the assay. Intra-assay and inter-assay coefficients of variation were in the ranges 5.3–7.1% and 4.8–13.3%, respectively. Accuracy, evaluated by adding 2 and 10 μg/ml of CRP to serum samples, provided recoveries of 99.9% and 106.8%. High correlation was found between CRP measurements by TR-IFMA and a by commercial enzyme-linked immunosorbent assay (R² = 0.98). The limit of detection for the TR-IFMA method was 0.000067 μg/ml and the measurement of CRP in serial dilutions of acute-phase dog sera generated curves with the same slope as the one constructed with purified CRP. The TR-IFMA provides a precise, accurate and highly sensitive assay for CRP determination in dog samples. CRP levels in dogs with different diseases ranged between 10.2 and 210.7 μg/ml and were significantly higher than those observed in healthy dogs (< 7.1 μmg/ml).     

Assessment of a pyrogallol red technique for total protein measurement in the cerebrospinal fluid of dogs

The measurement of protein concentration in the cerebrospinal fluid is a basic analytical method in neurology. In this study, a pyrogallol red technique using a human albumin calibrator previously validated in human medicine was tested for canine samples, and the results were compared with those obtained using urine test strips. Pyrogallol red significantly (P<0.05) but moderately underestimated purified dog albumin and globulins. The imprecision of the technique was low: intra- and between-series coefficients of variation were 1.6 and 4.3 per cent at protein concentrations of about 0.3 g/litre. Over 49 samples, there was good agreement between the pyrogallol red and test strip results (r=0.63), especially for low and high protein concentrations, but misclassifications were observed with '+' test strip readings.     

Assessment of three automated assays for C-reactive protein determination in dogs

OBJECTIVE: To determine the characteristics of an automated canine C-reactive protein (CRP) assay and evaluate 2 human CRP assays for use in dogs. Animals-56 client-owned dogs with pyometra and 11 healthy control dogs. PROCEDURES: Samples from 11 dogs with high (> 100 mg/L) or low (< 10 mg/L) CRP concentrations (determined by use of a canine ELISA) were evaluated by use of the automated canine CRP assay. Intra- and interassay imprecision was determined (by use of those 2 plasma pools), and assay inaccuracy was assessed by use of logistic regression analysis of results obtained via ELISA and the automated canine CRP assay. Two automated human CRP assays were used to measure plasma CRP concentration in 10 dogs. RESULTS: By use of the ELISA, mean +/- SD plasma CRP concentration was 96.1 +/- 38.5 mg/L and 10.1 +/- 23.2 mg/L in dogs with pyometra and control dogs, respectively. The automated canine assay had intra-assay coefficients of variation (CVs) of 7.8% and 7.9%, respectively, and interassay CVs of 11.1% and 13.1%, respectively. Results from the automated assay were highly correlated with results obtained via ELISA. The human assay results did not exceed 0.4 mg/L in any dog. CONCLUSIONS AND CLINICAL RELEVANCE: The automated canine CRP assay had less interassay imprecision, compared with the ELISA. The 2 human CRP assays were not suitable for analysis of canine plasma samples. The automated canine CRP assay was more precise than the ELISA for serial evaluations of plasma CRP concentration in dogs.     

A non-radiometric method for measuring serum thymidine kinase activity in malignant lymphoma in dogs

The aim of this study was to evaluate an enzyme-linked immunosorbent assay (ELISA), for determination of serum thymidine kinase 1 (sTK1) activity in dogs with malignant lymphoma (ML) and compare it with a thymidine kinase (TK) radioenzymatic assay (TK-REA). The TK-REA has recently been shown to be useful in determining the clinical stage and prognosis in canine ML. In addition, serum lactate dehydrogenase (LDH) was measured. Forty-five dogs were included in the study. Sixty serum samples from these dogs, stored in a tumour serum sample bank (stored at -20 degrees C), were analysed. Apart from 37 dogs with ML, four normal dogs as well as two dogs with mammary carcinomas, one dog with bladder carcinoma, and one dog with malignant fibrous histiocytoma were included. Staging of ML was based on the modified World Health Organization (WHO) staging system for canine ML. The diagnosis of all tumours was verified by histopathology. The TK activity (units per litre [U/L]) ranged from 1.0 to 607.9 in the TK-REA analysis and from 1.1 to 510 in the TK-ELISA (normal reference value <7U/L). The range for LDH was between 12 and 1194 U/L (normal reference value <228 U/L). There was a significant correlation between the TK-REA and the TK-ELISA. The correlation coefficient (CC) was 0.97 and the standard error of the estimate (SEE) was 3.7 U/L. There was no correlation between LDH and either the TK-REA or the TK-ELISA (CC=0.53 for both assays; SEE=26.7 and 12.7 U/L, respectively). Most of the variation in LDH was still within the normal reference range. The mean LDH in dogs with high-stage (stage IV+V) disease was 201.9 U/L. The corresponding values for the TK-REA and TK-ELISA were 109 and 109.9 U/L, respectively. The significant relation between the TK-REA and the TK-ELISA was confirmed by Bland-Altman analysis. The TK-ELISA assay, because of its relative simplicity, will permit measurement of TK in cases of ML in dogs to become a routine procedure.     

Evaluation of thyroid function in dogs suffering from recurrent flank alopecia

Thyroid function was assessed in euthyroid dogs (n = 20), dogs suffering from canine recurrent flank alopecia (CRFA, n = 18), and hypothyroid dogs (n = 21). Blood samples obtained from all dogs in each group were assayed for total thyroxine (TT4), thyrotropin (TSH), and thyroglobulin autoantibody (TgAA) serum concentrations. Total T4 and TSH serum concentrations were significantly decreased and increased, respectively, in the hypothyroid group compared with the other 2 groups. No significant differences in TT4 and TSH serum values were found between the euthyroid and CRFA groups. Thyroglobulin autoantibodies were detected in 10, 11.1, and 61.9% of euthyroid dogs, dogs with CRFA, and hypothyroid dogs, respectively. In conclusion, dogs suffering from CRFA have a normal thyroid function, and the determination of TT4 and TSH serum concentrations allows differentiation of these dogs from dogs with hypothyroidism, in most cases. Occasionally, the 2 diseases can be concomitant.     

Evaluation of a commercially available human C-reactive protein (CRP) turbidometric immunoassay for determination of canine serum CRP concentration

Background: Serum C-reactive protein (CRP) is an acute phase marker in dogs that is useful for the diagnosis and monitoring of inflammatory disease. Rapid, reliable, and automated assays are preferable for routine evaluation of canine serum CRP concentration.
Objective: The aim of this study was to evaluate whether canine serum CRP concentration could be measured reliably using an automated turbidometric immunoassay (TIA) designed for use with human serum.
Methods: A commercially available TIA for human serum CRP (Bayer, Newbury, UK) was used to measure canine serum CRP concentration. Cross-reactivity of antigen was evaluated by the Ouchterlony procedure. Intra-and interassay imprecision was investigated by multiple measurements on canine serum samples and serum pools, respectively. Assay inaccuracy was investigated by linearity under dilution and comparison of methodologies (canine CRP ELISA, Tridelta Development Ltd, Kildare, UK). Then the assay was applied to serum samples from 14 clinically healthy dogs, 11 dogs with neoplasia, 13 with infections, 8 with endocrine or metabolic diseases, and 10 with miscellaneous diseases.
Results: Cross-reactivity between canine serum CRP and the anti-human CRP antibody was found. Intra-and interassay imprecision ranged from 5.2% to 10.8% and 3.0% to 10.2%, respectively. Serum CRP concentration was measured in a linear and proportional manner. There was no significant disagreement and there was linear correlation of the results in the comparison of methodologies, except for a slight proportional discrepancy at low CRP concentrations (<10 μg/mL). Dogs with infections had a significantly higher concentration of serum CRP than did all other dogs, and dogs with neoplasia had a significantly higher concentration of serum CRP than did clinically healthy dogs.
Conclusions: Canine serum CRP concentration can be measured reliably using the commercially available TIA designed for human CRP.