A functional turbidimetric method to determine C-reactive protein in horses.

A turbidimetric method to determine serum C-reactive protein (CRP) concentration, based on soybean oil-phosphocholine interaction, was performed on horse serum samples to evaluate its potential diagnostic value in veterinary medicine. Intralipid 20% in 0.1 M Tris-calcium buffer (pH 7.5) was added to horse serum. After 30 min of incubation at 37 °C, the CRP-phosphocholine complexes were turbidimetrically, bichromatically (660 nm/700 nm) quantified on a commercial analyzer. Furthermore, comparison between CRP and other inflammatory markers, including white blood cell and neutrophil counts, was performed to evaluate the diagnostic value of both tests. Standardization of the assay was done using a commercial human CRP calibrator. The CRP measurements were performed on serum samples (296 patients and 34 controls). Reference values were found to be lower than 10 mg/l. The method was found to be linear between 1 and 400 mg/l. A moderate correlation was observed between CRP values and the relative neutrophil counts. Receiver-operating characteristics analysis demonstrated the area under the curve for CRP was 0.928, which was superior (P < 0.001) to the neutrophil count (0.804) and the leukocyte count (0.664) in detecting the presence of inflammation. This CRP assay showed reliable results as an acute phase test in horses, confirming its species-independent capability to detect CRP in various mammals, including horses.

A new turbidimetric method for assaying serum C-reactive protein based on phosphocholine interaction.

BACKGROUND: C-reactive protein (CRP) is able to bind phospholipids (mainly phosphocholine) in the presence of calcium ions. We investigated the use of this property for developing an affordable turbidimetrical CRP assay based on diluted soy oil. METHODS: Serum (or heparinized plasma) was mixed with Intralipid 20% in Tris-calcium buffer (pH 7.5). After 30 min of incubation at 37 degrees C, the CRP-phospholipids complexes were measured by turbidimetry (660 nm/700 nm) with a Cobas 6000 analyzer (Roche). Results were compared with those obtained using a typical immunoturbidimetric method (Roche). RESULTS: Good correlation (r(2)=0.931) was obtained between the functional and the immunoturbidimetric CRP assay. Within-run and between-run %CV values for the functional assay were 2.4% (100 mg/L); 6.0% (50 mg/L); 10% (20 mg/L), and 3.6% (100 mg/L); 8.0% (50 mg/L); 11% (20 mg/L), respectively. The limit of detection was 7 mg/L. Results were not affected by serum calcium, triglyceride, or phospholipid concentrations. CONCLUSIONS: The functional CRP assay allowed measurement of CRP in serum and plasma in the range of 7 mg/L-400 mg/L. The assay is particularly suited in conditions where resources are restricted. Since the assay is species independent, the described functional CRP assay could be used for veterinary purposes as well.