Comparison of quantitative immunoturbidimetric and semiquantitative latex-agglutination assays for D-dimer measurement in canine plasma.

BACKGROUND: D-dimer measurement in dogs is considered the most reliable test for detecting disseminated intravascular coagulation or thromboembolism. OBJECTIVES: The purposes of this study were to compare 2 D-dimer assays, a quantitative immunoturbidimetric and a semiquantitative latex agglutination assay, and to assess the effect of hemolysis and storage conditions on D-dimer concentration using the quantitative assay. METHODS: The immunoturbidimetric assay was validated using canine citrated plasma samples containing different concentrations of D-dimer. The effect of storage at various temperatures and times was assessed. Hemolysis was produced by adding lysed RBCs to the samples for a final hemoglobin concentration of 0.35 g/dL. RESULTS: For clinically relevant values (>250 microg/L), intra-assay and interassay coefficients of variation were 6.8% and 7.2%. The assay was linear (r(2)=1.00), and the tests had good agreement (kappa=0.685, P<.001). Storage at 4 degrees C and -20 degrees C and hemolysis had no significant effect on D-dimer concentrations. In hemolyzed samples stored at room temperature for > or =48 hours, fine clots were noted and often resulted in falsely increased D-dimer concentrations. CONCLUSIONS: Our findings suggest that the immunoturbidimetric assay validated in this study is reliable and accurate for the measurement of D-dimer in canine plasma. Samples can be stored for up to 1 month at -20 degrees C and moderate hemolysis does not significantly affect the D-dimer concentration in frozen or refrigerated samples.

Dynamics of bacterial growth and distribution within the liver during Salmonella infection.

Salmonella enterica causes severe systemic diseases in humans and animals and grows intracellularly within discrete tissue foci that become pathological lesions. Because of its lifestyle Salmonella is a superb model for studying the in vivo dynamics of bacterial distribution. Using multicolour fluorescence microscopy in the mouse typhoid model we have studied the interaction between different bacterial populations in the same host as well as the dynamic evolution of foci of infection in relation to bacterial growth and localization. We showed that the growth of Salmonella in the liver results in the spread of the microorganisms to new foci of infection rather than simply in the expansion of the initial ones. These foci were associated with independently segregating bacterial populations and with low numbers of bacteria in each infected phagocyte. Using fast-growing and slow-growing bacteria we also showed that the increase in the number of infected phagocytes parallels the net rate of bacterial growth of the microorganisms in the tissues. These findings suggest a novel mechanism underlying growth of salmonellae in vivo with important consequences for understanding mechanisms of resistance and immunity.