[Analysis of antibody reactivity for FDP D-dimer fragments by Western blotting].

Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Rinsho byori. The Japanese journal of clinical pathology 06/2008; 56(6):449-54.
Source: PubMed


We evaluated three test kits for fibrin degradation products (FDP) D-dimer. We found that six of 217 plasma sample values obtained by Nanopia test were markedly higher than the values obtained using the other two kits. The regression equation for 211 samples (excluding six) was y=0.64x+3.05 (y: Nanopia, x: LIAS AUTO) and the correlation coefficient was 0.915. Therefore, we classified these samples into three categories, namely correlated(y< 1.0x), incompatible (y= 1.0x-2.9x) and markedly incompatible (y> or =3.0x). Selected samples, eight correlated, four incompatible and four markedly incompatible, were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by Western blotting(WB). WB analysis using anti-fibrinogen antibody showed that both high molecular weight fragments of cross-linked fibrin (HMW-XDP) and DD/E fragments were present in the correlated samples, but there was less HMW-XDP than DD/E in the incompatible samples and mostly DD/E (HMW-XDP was significantly less than DD/E) in the markedly incompatible samples. These data suggest that plasma FDP samples that contain mostly DD/E and little HMW-XDP demonstrated markedly incompatible values using the three D-dimer test kits. These data was reflected by markedly elevated plasmin alpha2-plasmin inhibitor complex values in the incompatible and markedly incompatible samples. Unfortunately, we did not directly demonstrate these phenomena by WB analysis with two anti-D-dimer antibodies used Nanopia or LPIA reagent. In the near future, we expect that standardization of FDP D-dimer assay will be accomplished.

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    • "D-D reagents from various manufacturers have different normal values. Manufacturers prepare different D-D molecular weight degradation fragments with various monoclonal antibodies, thus, D-D has highly variable weights, ranging between 50 and >228,000 Da (14). In the present study, D-D concentrations were shown to be significantly higher in patients with acute cerebral infarction as compared with the controls (P<0.01). "
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    ABSTRACT: Cerebral infarction is usually associated with arteriosclerosis, vascular endothelial cell injury and blood flow through the vascular system. The diagnostic value of markers, including von Willebrand factor antigen (vWF:Ag), D-dimer (D-D) and fibrinogen/fibrin degradation product (FDP), have not been studied in patients with acute cerebral infarction. Thus, the aim of the present study was to use receiver operating characteristic (ROC) curves to evaluate the diagnostic significance of vWF:Ag, D-D and FDP in 94 cases of acute cerebral infarction. The results revealed that vWF:Ag and D-D concentrations were significantly higher in acute cerebral infarction patients as compared with the normal controls (P<0.01), whereas no statistically significant difference in FDP was observed between the groups (P>0.01). Plasma vWF:Ag and D-D concentrations significantly correlated with the National Institute of Health Stroke Scale (NIHSS) scores (r=0.625 and 0.582, respectively; P<0.01). In addition, the vWF:Ag concentration significantly correlated with the D-D concentration (r=0.320; P<0.01), whereas FDP concentration did not correlate with D-D or vWF:Ag concentrations or the NIHSS scores (r=0.172, 0.188 and 0.065, respectively; P>0.05). The area under the ROC curve using vWF:Ag as a diagnostic marker in patients with acute cerebral infarction was 0.900, while for D-D the area was 0.795 and for FDP the area was 0.465. Logistic regression analysis revealed that the odds ratios of vWF:Ag and D-D were 16.727 and 2.324, respectively, which were statistically significant (P<0.001 and 0.023, respectively). These results indicated that using vWF:Ag as a diagnostic marker is likely to significantly improve the sensitivity of diagnosing patients with acute cerebral infarction. The diagnostic value of vWF:Ag concentration was significantly higher compared with D-D and FDP levels.
    Experimental and therapeutic medicine 06/2014; 7(6):1573-1577. DOI:10.3892/etm.2014.1665 · 1.27 Impact Factor