[Analysis of antibody reactivity for FDP D-dimer fragments by Western blotting].
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.
Wakabayashi
Saki
S
Department of Laboratory Medicine, Shinshu University Hospital, Matsumoto, Japan.
Ishikawa
Shinsuke
S
Hirota-Kawadobora
Masako
M
Fujihara
Noriko
N
Kamijyo
Yuka
Y
Yamauchi
Kazuyoshi
K
Terasawa
Fumiko
F
Okumura
Nobuo
N
Katsuyama
Tsutomu
T
D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis / Q000276:immunology*; D006801:Humans; D011933:Reagent Kits, Diagnostic
D000911:Antibodies, Monoclonal; D005338:Fibrin Fibrinogen Degradation Products; D011933:Reagent Kits, Diagnostic; C036309:fibrin fragment D