Monoclonal antibody to fibrin D-dimer (DD-3B6) recognizes an epitope on the gamma-chain of fragment D

Laboratory determination of fibrinolysis has been facilitated by diagnostic tests that use monoclonal antibody DD-3B6 to measure fibrin D-dimer levels in plasma. When DD-3B6 is reacted with soluble fibrin fragments, it is specific for fragment D-dimer. The authors have used immunoblot analysis of DD-3B6 binding to purified fragment D and fragment D-dimer to localize the binding site of the antibody. Although DD-3B6 recognizes only fragment D-dimer in solution, it binds to both immobilized fragment D-dimer and fragment D in immunoblots. When immunoblots were performed using protein which was electrophoresed under reducing conditions, DD-3B6 bound to the gamma-chain of fragments D1, D2, and D3. Therefore, the epitope recognized by DD-3B6 resides between amino acids 86 and 302 in the gamma-chain of fragment D. This epitope is masked in soluble non-cross-linked or nondegraded fibrin, but becomes expressed after cross-linked fibrin has been cleaved by plasmin.     

Measurement of plasma D-dimer for diagnosis of deep venous thrombosis

Venography was performed on fifty-six patients suspected of having deep venous thrombosis (DVT) of the legs. The accuracy of the D-dimer measurement in plasma using two latex tests and an enzyme-linked immunosorbent assay (ELISA) was compared with that of usual determination of total fibrin(ogen) degradation products (FDPs) in serum with respect to the presence of DVT. The three D-dimer tests were clearly superior to the FDP assay, but only the ELISA could accurately rule out the diagnosis of DVT with a predictive value of 100% when plasma D-dimer level was less than 200 micrograms/L. However, this test cannot be used for positive diagnosis (false positive rate of 69%). Thus, plasma D-dimer measurement with ELISA allows identification of patients in whom further investigation by means of more specific tests (venography or plethysmography) is indicated in order to establish the diagnosis of DVT. In contrast to this, sensitivity of the two latex tests studied was low (60 and 76%, respectively), which makes them unsuitable for emergency screening. In addition, the potential of D-dimer dosage for diagnosis of DVT in hospitalized patients is hampered by the presence of associated conditions that are responsible for elevated plasma levels in most cases.     

Diagnosis of disseminated intravascular coagulation. Role of D-dimer

Detection of the cross-linked fibrin degradation fragment, D-dimer, in patients at risk for disseminated intravascular coagulation (DIC) is strong evidence for the diagnosis. D-dimer confirms that both thrombin generation and plasmin generation have occurred. Patients at risk for DIC (58) and normal controls (7) were studied. Thirty-three patients had DIC--with fragment D-dimer identified in their serum by immunoblotting. Latex agglutination measurements of fibrin(ogen) degradation products (FDPs) and D-dimer were compared with immunoblotting in the detection of D-dimer. FDP measurement was extremely sensitive but not specific. D-dimer measurement was less sensitive but highly specific. Used in tandem, screening with FDP and confirming with D-dimer, sensitivity and specificity were maximized, rendering a predictive value of a confirmed FDP of 100% in this cohort. D-dimer is a valuable adjunct for the laboratory diagnosis of DIC but is most appropriately used as a confirmatory test for the very sensitive FDP test.     

Rapid detection of cross-linked fibrin degradation products in plasma using monoclonal antibody-coated latex particles

Conformational and structural changes on conversion of fibrinogen to fibrin and its cross-linking by Factor XIIIa lead to the development of new antigenic determinants that permit differentiation between their plasminolytic cleavage products. A monoclonal antibody (DD-3B6/22) that is specific for cross-linked fibrin derivatives containing the D dimer configuration has been used in developing a latex agglutination procedure that can detect fibrin degradation products in either plasma or serum. Fibrinogen or its degradation products do not cross-react with this antibody. Results were calibrated with an enzyme immunoassay, which used a purified D dimer standard. Plasmas from 40 normal subjects, all having D dimer levels below 250 ng/mL measured by enzyme immunoassay, were all negative by latex assay. In contrast, positive latex agglutination titers were obtained with 87 of 88 patients with demonstrated deep venous thrombosis, pulmonary embolism, or disseminated intravascular coagulation. Compared to enzyme immunoassay, latex agglutination assay is less sensitive, but this latex procedure provides a rapid and less elaborate test for elevated levels of cross-linked fibrin degradation products in patients with thrombosis. Plasma assays for fibrin degradation products are preferable to those using serum.     

False-positive D-dimer result in a patient with Castleman disease

In suspected cases of disseminated intravascular coagulation, concurrent elevation of both fibrin(ogen) degradation products (FDPs) and D-dimer levels aids in confirming the diagnosis. This pattern of results reflects the action of plasmin proteolysis of cross-linked fibrin polymers as well as fibrinogen. We report the case of a patient with human immunodeficiency virus (HIV) and Castleman disease who presented with a high-positive D-dimer level and a negative FDP level in the course of a workup for disseminated intravascular coagulation. This finding suggested the possibility of either a false-positive D-dimer or a false-negative FDP level. To investigate the former, a Western blot was performed on the patient's serum to determine the presence of the D-dimer. No D-dimer band was visualized on the Western blot, confirming the false-positive nature of the D-dimer result. Insufficient quantity of patient serum, however, prevented further investigation into the etiology of this result. The false-positive D-dimer result is likely attributable to interference caused by the patient's Castleman disease-associated monoclonal gammopathy, a phenomenon that has been reported in other immunoassays. As the development of lymphoproliferative disorders is especially common within the HIV population, and hypergammaglobulinemia in Castleman disease is particularly common, clinicians should be aware of this phenomenon when the laboratory findings do not fit the clinical picture. Although it is rare, recognition of potential paraprotein interference in immunoassays will help avoid undertreatment or overtreatment of patients based on erroneous laboratory results.     

Clinical significance of a new fibrinogen/fibrin degradation products(FDP) test using plasma samples for the diagnosis of fibrinogenolysis and fibrinolysis

Recently, a new fibrinogen/fibrin degradation products(FDP) test using monoclonal antibodies against FDP(LPIA FDP-P: FDP-P) has been developed, which is able to measure FDP directly in plasma. The objective of this study is to clarify clinical significance of the test in the diagnosis of fibrinogenolysis and fibrinolysis in comparison with a conventional FDP test using polyclonal antibodies against fibrinogen(FDP-S) and D-dimer test using monoclonal antibodies against D-dimer(D-D). The monoclonal antibodies used in FDP-P test was shown to recognize fragment X, Y and D1 derived from fibrinogen digested by urokinase, and was also to recognize XDP fragments, D-dimer and D derived from cross-linked fibrin digested by tissue plasminogen activator using SDS-PAGE and immunoblotting analysis. There was a good correlation of FDP levels between FDP-P test and FDP-S test. However, levels of FDP in both tests were discrepant in several samples. There was a tendency that the levels of FDP were higher in FDP-S test than in FDP-P test. Such discrepancy was suggesting that soluble fibrin monomer complex(FM) was recognized by the antibodies used in FDP-S test, but not recognized by the antibodies used in FDP-P test. There was also a good correlation of FDP levels between FDP-P test and D-D test. However, the levels of FDP in both tests were discrepant in several samples. The levels of FDP were higher in FDP-P test than in D-D test. These discrepant samples had lower levels of antiplasmin and higher levels of plasmin antiplasmin complex(PIC), and also showed XDP fragments, D-dimer, X, Y, and D1 by using SDS-PAGE. These observations suggest that D-D test measures only fibrinolytic fragments, while FDP-P test measures fibrinogenolytic fragments as well as fibrinolysis. In results, the FDP-P test was confirmed to be a useful tool to examine fibrinogenolysis as well as fibrinolysis more specifically than the conventional FDP test.     

Fibrinolysis in idiopathic menorrhagia.

Serum fibrin/fibrinogen degradation product (FDP) was studied in 64 patients of menorrhagia without any organic cause in addition to 24 healthy women by Thrombo - Wellcotest HA - 13 Kit (Wellcome, England). Serum FDP levels were found to be less than 10 micrograms/ml in healthy subjects, whereas in idiopathic menorrhagia it was more than 10 micrograms/ml in 59.34% patients. Semi-quantitative estimation of FDP in 14 patients of idiopathic menorrhagia indicated a positive correlation between duration of bleeding and FDP levels. Bleeding appears to be due to increased fibrinolytic activity in uterus secondary to plasminogen activator. Such patients are likely to be benefitted with anti-fibrinolytic agents.     

Degradation products of fibrin and of fibrinogen in synovial fluid and in plasma of patients with rheumatoid arthritis

Degradation products of fibrin and of fibrinogen, together (FDP) and separately, were measured in plasma and synovial fluid of patients with rheumatoid arthritis using specific enzyme immunoassays. Considerable amounts of fibrin degradation products (fibrin-DP) were found in the synovial fluid; only 1 case yielded some fibrinogen-derived products (fibrinogen-DP). In plasma relatively small amounts of degradation products, both fibrin-DP and fibrinogen-DP, were present. From the discrepancy between the amount and composition of degradation products in synovial fluid and in plasma we conclude that there are extra-articular sites of fibrin(ogen) breakdown in rheumatoid arthritis. The new specific methods may contribute to the understanding of the role of fibrinolysis in rheumatoid disease and in inflammation in general.     

Incidence and possible reasons for discordant results between positive FDP and negative D-dimer latex assays in clinical specimens.

In general, FDP and D-dimer values have a correlation in clinical conditions associated with disseminated intravascular coagulation(DIC) or coagulation activation. However, there are some patients with discordant results who demonstrate elevated FDP and negative D-dimer results by latex agglutination assays. The incidence and possible reasons for the discordance between FDP and D-dimer results were investigated through simultaneous measurements (n = 763) from clinical patients with suspected DIC or coagulation activation. 24.8% (189/763) of samples with elevated FDP were negative for D-dimer assays by the latex agglutination method. Further detailed analysis on randomly-selected discordant samples (n = 41) revealed that the most common reason for the discordance was the lower sensitivity of the semiquantitative latex agglutination method for D-dimer, compared with quantitative enzyme or other latex immunoassay. The other contributing factors to the discordance were accelerated fibrinogenolysis without secondary fibrinolysis, elevated soluble fibrin monomer and rheumatoid factor.     

30例D-二聚体假性增高的数据分析

目的探讨30例临床检测中血浆D-二聚体水平>FDP水平的原因及相关资料分析。方法收取临床检测过程中首次出现血浆D-二聚体水平>FDP水平的标本30例。分别用血浆D-二聚体检测试剂(STA-LIATEST D-DI和STA-LIATEST D-DI PLUS)检测。并对其进行数据比较和原因分析。结果血浆D-二聚体检测试剂STA-LIATEST D-DI与STA-LIATEST D-DI PLUS数据相比较,[3.63(2.89~10.49)μg/mL vs 0.36(0.26~1.00)μg/mL,P<0.0001],两组数据的差别具有统计学意义。30例患者中21例患者类风湿因子(rheumatoid factor,RF)水平异常,>20 IU/mL(正常参考值上限)。9例患者类风湿因子水平正常,<20 IU/mL。结论血浆D-二聚体检测试剂STA-LIATEST D-DI PLUS对于D-二聚体水平假性增高的标本有很强的的纠正能力。血浆D-二聚体水平>FDP水平时,绝大多数是由于类风湿因子干扰造成的,少部分原因不明确,可能由其他异嗜性抗体导致。     

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.     

Fibrin monomer assay

Quantitative assay for fibrin monomer was done by use of a chromogenic substrate (S-2390, Coa set fibrin monomer). Samples from DIC prone patients with the underlying disease were assayed and classified into four groups. The pre DIC group showed higher FM values than the control with no laboratory coagulation abnormality, although the FDP . D-dimer showed no significant rise. FM assay is a useful marker for the detection of early coagulopathy in DIC. Administration of the AT III concentrate in the case of low level of plasma ATIII, thrombin . antithrombin complex I (TAT) caused a significant transient rise. The clinical course of DIC by TAT is often affected by the fluctuation of ATIII level in plasma, the usefulness of FM is that it reflects the real thrombin generation in DIC.     

Clinical usefulness of the measurement of plasma D-dimer levels

To evaluate the clinical usefulness of D-dimer, various effects on the measurement of D-dimer were examined. Although both fibrinolytic and fibrinogenolytic products were detected by the measurement of FDP, only fibrinolytic products were detected by the measurement of D-dimer. In patients with DIC and other thrombo-embolic diseases, plasma D-dimer levels were significantly higher than in normal persons. A significant positive correlation between plasma D-dimer and serum FDP was found in DIC patients. In patients with DIC associated with acute promyelocytic leukemia, which is thought to be an increased fibrinogenolysis state, serum FDP was higher than the plasma D-dimer which suggests that increased fibrinogenolysis affects the result of serum FDP measurement. Plasma D-dimer significantly increased 5 minutes after endoscopic embolization with thrombin in the patients with esophageal varices. However serum FDP increased 30 minutes after the treatment, which suggests that the D-dimer is more useful for rapid detection of coagulo-fibrinolytic change than serum FDP. Plasma D-dimer was significantly higher in patients with cerebral infarction and increased with age. These finding suggest the usefulness of plasma D-dimer measurement for the specific and rapid evaluation of coagulo-fibrinolytic activation and thrombo-embolic state.     

Diagnostik der disseminierten intravasalen Gerinnung: Aussagekraft von löslichem Fibrin,D-Dimeren und Fibrin(ogen)-Spaltprodukten

Summary The validity of the fibrin(ogen) derivatives soluble fibrin, D-dimers and fibrin(ogen) degradation products was compared with other parameters in early and sensitive diagnosing of disseminated intravascular coagulation (DIC). In a clinical study 900 patients' samples from separate, defined groups were examined, including course observations of intensive care patients (n=38) and patients with acute pancreatitis. The fibrin(ogen) derivatives correlated very well with the degree of blood coagulation disturbances: in particular, D-dimers and soluble fibrin proved to be more sensitive in early diagnosis of DIC than other parameters. The SF-PS-turbidimetry demonstrated a good validity and practicality in the quantitative determination of soluble fibrin, but a suitable analyzer is essential. Determination of D-dimers is preferable to that of fibrin(ogen) degradation products (both in the latex-agglutination test) because of the better sensitivity and practicality; even more sensitive results were provided by the D-dimer-ELISA, which is, however, not practical in acute diagnostics. The decrease in protein C was at least equally sensitive as the antithrombin III-levels in indicating the consumption of the hemostatic potential. The decrease of thrombocyte counts and fibrinogen levels could first be detected in a later stage of DIC.In conclusion, D-dimers and soluble fibrin can improve the DIC diagnostics, making them more reliable; additionally, antithrombin III and possibly protein C deserve further consideration, although the fibrin(ogen) derivatives are apparently of greater importance.

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Abkürzungsverzeichnis ATIII Antithrombin III - DIC disseminierte intravasale Gerinnung - ELISA Enzyme-linked Immuno Sorbent Assay - FII, FV, FXIII Gerinnungsfaktor II, V, XIII - FDP Fibrin(ogen)-Spaltprodukte - FPA Fibrinopeptid A - LAT Latex-Agglutinationstest - PS Protaminsulfat - PTT partielle Thromboplastinzeit - tPA Gewebeplasminogen-Aktivator - TAT Thrombin-AntithrombinIII-Komplex - TPZ Thromboplastin-Zeit     

Measurement of cross linked fibrin derivatives in plasma: an immunoassay using monoclonal antibodies.

Fibrinogen degradation, fibrin polymerisation, and the insertion of cross links into fibrin by fibrin stabilising factor lead to the appearance of new antigenic determinants. Antibodies against these antigenic sites may react specifically with the derivatives but not with the parent molecules. We have utilised a monoclonal antibody, which interacts with the cross linked fragment D dimer and related high molecular weight fibrin derivatives, to develop an enzyme immunoassay which measures cross linked fibrin derivatives in plasma and serum using D dimer as standard. Mean concentration in plasma from normal subjects was 75 ng/ml with an upper limit of about 144 ng/ml. Concentrations in patients with pulmonary embolism, deep venous thrombosis, arterial thromboembolism, and disseminated intravascular coagulation were raised in all cases. Confirmation of the specific increase of cross linked fibrin derivatives in patients with disseminated intravascular coagulation was obtained by parallel monitoring of their fibrin degradation products in serum using affinity chromatography and sodium dodecyl sulphate (SDS) polyacrylamide gel electrophoresis. In many patients the plasma concentrations greatly exceeded the serum values of cross linked fibrin degradation products, suggesting that the procedure can measure fibrin derivatives in plasma which are absent from serum.     

Coagulation and fibrinolysis in pregnancy]

We studied on coagulation and fibrinolysis systems during pregnancy by measuring plasma levels of thrombin-antithrombin III complex (TAT), plasmin-alpha 2 plasmin inhibitor complex (PIC), fibrinogen/fibrin degradation products (FDP), plasminogen (PLG) and antithrombin III (AT-III). Ninety seven pregnant, 5 post-delivery and 32 nonpregnant women aged from 20 to 52 years old were included in this study. Plasma concentrations of TAT and PIC in nonpregnant women were 3.38 +/- 1.02 micrograms/l and 0.65 +/- 0.24 micrograms/ml, respectively. TAT gradually increased with the progression of pregnancy and rapidly decreased after the delivery. Whereas PIC and AT-III concentrations did not change significantly during pregnancy. Fibrinogen, PLG and FDP concentrations changed similarly as TAT. Eight pregnants whose plasma PIC concentrations elevated more than 1.0 micrograms/l were further examined. In 6 women out of them (71.5%), FDP concentrations were elevated. In this particular group of subjects, however, they delivered normally without complications such as toxemia. These observations suggest that, at least, a hypercoagulative state progresses with pregnancy, being normalized after the delivery. Although we could not find the relationship between the hypercoagulation and clinical complications such as thrombosis and toxemia of pregnancy, present findings suggest that special caution should be paid on the pregnants whose TAT and FDP levels are elevated.     

A case of hookworm infestation with dissociation values between FDP-E and FDP-D dimer

We previously reported a five-year-old girl showing bleeding tendency and transient morphological and functional platelet abnormalities probably due to a hookworm, Necator Americanus, infestation. In this report, we describe the rarely accelerated fibrinogenolysis and/or fibrinolysis in this patient whose value of fibrinogen and/or fibrin degradation products(FDP) determined with an FDP-E assay was much higher than that determined with a D-dimer assay. Namely, on day-1 and day-13 of hospitalization, her D-dimer values were only 10 to 20% of the prospected values from FDP-E values. We speculated this phenomenon was induced by circulating protease(-like) agent(s) produced by hookworm, because the only slightly participation of plasmin and/or granulocyte elastase was evaluated by the determination of enzyme-inhibitor complexes. And the other possibility of fibrinogen degradation by blast- or tumor-associated protease was excluded by the clinical manifestations and primary disorders. In conclusion, we report a very rare case with the accelerated fibrinogenolysis and/or fibrinolysis in a patient with the hookworm infestation. We are interested in the mechanism that manifested the patient's bleeding tendency accompanied with morphological and functional platelet abnormalities.     

Clinical application of laboratory diagnosis: leukemia and DIC]

Plasma levels of molecular markers of hemostatic activation were investigated in 205 samples from patients with haematopoietic malignancies. These markers included thrombin/antithrombin III complex (TAT), D-dimer, plasmin/alpha 2plasmin inhibitor complex (PIC) and thrombomodulin (TM), and were assayed by EIA methods. Samples were divided into 4 groups according to the level of FDP: group A; FDP 10 greater than, group B; 10 less than or equal to less than 20 group C; 20 less than or equal to less than 40, and group D; less than 40. The mean level of each marker except TM increased in the order of group A, B, C and D. However, in many samples belonging to group A the plasma TAT or PIC levels and both were increased in spite of low FDP level. Furthermore, levels of TAT and PIC in several samples belonging to groups C and D were within the normal range. Also, the mean levels of each marker except TM increased in the order of 2, 3, 4, 5 and over 6 points in DIC score according to the criteria of DIC diagnosis by the research committee on DIC of the Ministry of Health and Welfare in Japan. Eight of the 11 samples (72.7%) obtained from cases with a DIC score of 3 points had high plasma levels of TAT, PIC and D-dimer. Plasma levels of these markers were increased after chemotherapy. These findings lead to the following conclusions: 1) FDP reflexed activation of coagulation and fibrinolysis, but 2) FDP was not more sensitive than TAT and PIC, and 3) the increase of FDP rarely resulted from fibrinogenolysis or non-plasmin mediated fibrinolysis. Furthermore, 4) TAT, D-dimer and PIC may serve as sensitive parameters of hemostatic activation in circulating blood and be valuable markers for early diagnosis of DIC.     

Prevalence and clinical implications of heparin-associated false positive tests for serum fibrin(ogen) degradation products

An electrophoretic method has been applied to characterize specific fibrinogen and fibrin degradation products (FDP) in 135 serum samples from 59 consecutive patients having a positive latex agglutination test for serum FDP in the evaluation of consumption coagulopathy. In 20 of 135 positive samples, the principal fibrinogen derivatives present were not degradation products of fibrinogen or fibrin but were instead residual fibrinogen or fibrin monomer and polymers (FFMP) due to incomplete clotting. Heparin exposure was common in patients with positive FDP tests occurring in 29 of 59 patients (49%) with 81 of 135 samples (60%). Heparin exposure by parenteral administration or catheter was significantly correlated with a false positive serum FDP test because of residual FFMP occurring in 19 of 81 (23%) samples from heparin-exposed patients but in only 1 of 54 (2%) samples from patients without exposure (P less than 0.005). Treatment of the false positive samples with reptilase, an enzyme unaffected by heparin, resulted in complete removal of the residual FFMP, and in vitro experiments demonstrated that heparin-containing plasma samples could be completely clotted with either reptilase or protamine sulfate plus thrombin. Survey of 20 regional laboratories showed that only 10% used reptilase or protamine sulfate to prepare serum if heparin exposure had occurred and that this was done in only 22 of 5,049 (0.4%) of samples in the last calendar year. Greater attention should be given to proper preparation of serum from heparin-exposed patients, and physicians should be aware of the possibility of falsely positive or falsely elevated serum FDP tests in evaluation of consumption coagulopathy in heparin-exposed patients.     

Clinical evaluation of a test for plasma fibrin/fibrinogen degradation products (FDP) based on monoclonal anti-FDP antibody technology: an application for the scoring system of the disseminated intravascular coagulation (DIC) diagnostic criteria

Fibrin/fibrinogen degradation products(FDP) have been measured using serum samples which were specially prepared for the FDP test because of the usage of anti-human fibrinogen antibody for the assay. Since diagnostic criteria for DIC were established by the study group on thrombosis and hemostasis which is supported by the Japanese Ministry of Health and Welfare(JMHW), serum FDP assay have been used as standard methods to diagnose DIC in Japan. Recently, a reagent using an anti-human FDP monoclonal antibody was developed and this has enabled the use of plasma samples for FDP measurement. The comparability, especially of the DIC score, of a new assay, Latex test BL-2 P-FDP, using plasma samples with a conventional assay for serum was investigated. Two sets of DIC scores based on data from the two tests were compared and the correlation was high with 97.5% of the patients being diagnosed with the same DIC status. In four disease groups--DIC, thrombosis, leukemia and solid cancer--high comparability between the two tests was also shown and no significant difference was observed in the correlation coefficient and the slope coefficient between serum and plasma samples. To conclude, it is suggested that "Latex test BL-2 P-FDP" is applicable to the diagnostic criteria for DIC from JMHW without any difficulty.