Abstract C-reactive protein (CRP) is able to bind phosphocholine in the presence of calcium ions. According to a previous functional property of CRP, we tried to develop an affordable and cheap high sensitive nephelometric CRP assay using soy oil. Serum samples were measured by Nephelometer BNII (Siemens), by mixing the serum with diluted soy oil emulsion (Intralipid ® 20%) and Tris-calcium buffer (PH 7.5). The measurement took place after 12 min incubation time at 37°C by measuring the agglutination between CRP and phosphocholine. Results from our automated functional assay were compared with results obtained using an immunoturbidimetric CRP assay. Results showed a good correlation coefficient for method comparison between functional nephelometric CRP assay and immunoturbidimetric CRP assay, r = 0.895, significance level p <0.0001. The limit of detection for the functional nephelometric CRP assay was 0.1 mg/L. However, the within run % CV values for the functional assay were 6.1 % (20 mg/L), 4.7 % (50 mg/L) and 4.5 % (100 mg/L). The between-run % CV values were 17.6 % (20 mg/L), 18.8 % (50 mg/L), and 11.3 % (100 mg/L). The new functional nephelometric CRP assay enables high sensitive CRP measurement in serum in the range of 0.1 mg/L to 300 mg/L. The functional assay could be used for veterinary analysis due to the ability to measure CRP according to the functional properties, not the morphological properties which depend on specific antibodies.

An inflammatory reaction is induced after release of proinflammatory mediators such asinterleukin 1 and 6 and tumour necrosis factor α. These mediators stimulate the liver tosuppress the syntheses of albumin and endure the syntheses of acute phase protein forinstance C-reactive protein. The aim of this paper was to perform a method validation on animmune turbidimetric assay to quantify C-reactive protein in canine serum at the laboratory atSkara Animals Hospital, Skara, Sweden. The validation involved evaluation of the assaylinearity, precision, stability and recovery.The method was proved to be linear for both TruLab control and Medinor control. TheTruLab control is based on human C-reactive protein while the Medinor control is based oncanine C-reactive protein. The precision of the method had a CV of 2.26 % (n=40). Themethod is considered stable and has good precision, under the circumstance that the TruLabcontrol is used. It was concluded that a Canine CRP-control is required and should beincluded in routine analysis.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2014-11-19T12:58:15Z No. of bitstreams: 2 Dissertação - Letícia Rodrigues Carneiro - 2013.pdf: 1923429 bytes, checksum: f5e2e5e1811c92e7dd1e953454554af4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-11-20T14:23:59Z (GMT) No. of bitstreams: 2 Dissertação - Letícia Rodrigues Carneiro - 2013.pdf: 1923429 bytes, checksum: f5e2e5e1811c92e7dd1e953454554af4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-11-20T14:23:59Z (GMT). No. of bitstreams: 2 Dissertação - Letícia Rodrigues Carneiro - 2013.pdf: 1923429 bytes, checksum: f5e2e5e1811c92e7dd1e953454554af4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-12-06 / Acute phase proteins contribute to homeostasis, limit bacterial growth and, more recently, have been used as biochemical markers of nutritional status. This study aimed to determine the concentrations of acute phase proteins in dogs with different body condition scores (BCS). We used a 1-9 scale to determine the body condition score, established by inspection and palpation. We evaluated 60 dogs of different breeds and ages, being 20 lean animals (10 males and 10 females – BCS 3), 20 with ideal body score (10 males and 10 females – BCS 4 to 6), and 20 obese animals (10 males and 10 females - BCS 7 to 9). The animals fasted for 12 hours before blood sample collection for laboratory evaluation. We measured CBC, fibrinogen quantification, glucose, total protein, albumin, globulin, urea, cholesterol, triglycerides, iron, iron binding capacity (IBC), transferrin saturation index (STI), transferrin, ferritin determination, and serum protein electrophoretic fractionation on agarose gel. Leukocytes, fibrinogen, glucose, total protein, globulin, cholesterol, triglycerides, iron, IST, transferrin, alpha-1, alpha-2, beta-2 and gamma globulin showed significant difference among groups (p < 0.05). Regarding acute phase proteins, we concluded that obesity in dogs determines an increase in fibrinogen and alpha-2, beta-2 and gamma globulin fractions, indicating chronic inflammation. Higher values of leukocytes, total protein, globulin, iron, IST, glucose, cholesterol and triglycerides are expected in hematological and metabolic profile in obese dogs. / As proteínas de fase aguda contribuem para homeostasia, limitam o crescimento bacteriano e, mais recentemente, passaram a ser utilizadas como marcadores bioquímicos do estado nutricional. Com este estudo objetivou-se determinar as concentrações das proteínas de fase aguda em cães com diferentes escores de condição corporal (ECC). Na determinação do escore de condição corporal foi utilizada escala de 1 a 9, estabelecida por palpação e inspeção. Foram avaliados 60 cães de diferentes raças e idades, sendo 20 com ECC abaixo do ideal, 20 com ECC ideal e 20 animais com ECC acima do Ideal, em cada grupo, sendo 10 machos e 10 fêmeas. Para as colheitas das amostras sanguíneas para avaliação laboratorial os animais estavam em jejum de 12 horas. Foi realizado hemograma, quantificação do fibrinogênio, determinação da glicose, proteína total, albumina, globulina, ureia, colesterol, triglicerídeos, ferro, capacidade de ligação do ferro (IBC), índice de saturação da transferrina (IST), transferrina, ferritina e fracionamento eletroforético das proteínas séricas em gel de agarose. Apresentaram diferença significativa entre grupos (p<0,05) os leucócitos, proteína total, globulina, glicose, colesterol, triglicerídeos, ferro, IST, fibrinogênio, transferrina, alfa-1, alfa-2, beta-2 e gama globulina. Em relação às proteínas de fase aguda pode-se concluir que a obesidade em cães determina elevação do fibrinogênio e das frações globulínicas alfa-2, beta-2 e gama, indicando presença de processo inflamatório crônico. No perfil hematológico e metabólico de cães obesos são esperados valores mais elevados de leucócitos, proteína total, globulina, ferro, IST, glicose, colesterol e triglicerídeos.