Determination of heparin‐induced IgG antibody in heparin‐induced thrombocytopenia type II

Wang, L. C.; Huhle, G.; Malsch, Reinhard; Hoffmann, V; Song, Xiaoming; Harenberg, Job

doi:10.1046/j.1365-2362.1999.00433.x

Cited by 13 publications

(7 citation statements)

References 22 publications

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“…Whether this finding can also explain the lower incidence of HIT in danaparoid‐treated patients remains to be studied in a larger series of experiments (Magnani, 1993). To date, the lower incidence of HIT in these patients has been attributed to the difference in the chemical structure of danaparoid and the heparins (Magnani, 1993; Greinacher et al , 1994; Chong, 1995) and a lower incidence of the binding of danaparoid to HIT‐IgG in the presence of platelet factor 4 (Wang et al , 1999b). The present data support the literature findings whereby LMW heparins crossreact with HIT antibodies in 100% of patients.…”

Section: Discussionmentioning

confidence: 99%

Determination of serotonin release from platelets by enzyme immunoassay in the diagnosis of heparin‐induced thrombocytopenia

Harenberg¹,

Huhle²,

Giese³

et al. 2000

Br J Haematol

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Summary. [14 C]-Serotonin release assay (14C-SRA) from platelets is considered to be the most sensitive test for laboratory confirmation of heparin-induced thrombocytopenia (HIT). This study compared 14C-SRA with an enzyme immunoassay (EIA) to determine the release of serotonin from platelets in the presence of heparin and serum from HIT patients. The normal range (median, 2´5 and 97´5 percentiles) of serotonin release from platelets in healthy subjects (n 149) is 38 ng/ml (19 and 62) measured by EIA-SRA. Serum from HIT patients (n 42) released 2548 ng/ml (244 and 7987) serotonin in the presence of 0´1 IU/ml heparin and 29 ng/ml (13 and 76) in the presence of 100 IU/ml heparin. One hundred per cent and 15% of HIT samples exhibited an elevated serotonin release from platelets in the presence of 0´1 IU/ml low molecular weight (LMW) heparin, 2100 ng/ml (869 and 5968), or danaparoid, 272 ng/ml (143 and 403), respectively. The sensitivity and specificity of the EIA-SRA was 100% and 97´4% and of the 14C-SRA 100% and 92´9% in HIT patients. No falsepositive results were found in patients receiving heparin (n 28), in patients with elevated levels of bilirubin (n 5), in patients with antiphospholipid antibody syndrome (n 10) or in non-HIT patients (n 78) with both assays. The EIA technique to quantify serotonin release from platelets provides a reliable non-radioactive method to diagnose heparin-induced thrombocytopenia and to assess in vitro crossreactivity of low molecular weight heparins and heparinoid.

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Section: Discussionmentioning

confidence: 99%

Determination of serotonin release from platelets by enzyme immunoassay in the diagnosis of heparin‐induced thrombocytopenia

Harenberg¹,

Huhle²,

Giese³

et al. 2000

Br J Haematol

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“…This membrane‐based assay detected IgG antibodies in 73 SRA‐positive HIT sera, in which anti‐PF4–heparin antibody was detected. In a study using an assay based on capture of HIT antibody‐hPF4–heparin by protein A, Wang et al (1999) found HIT IgG in 37 out of 38 HIT sera. In contrast, using ELISA with covalently bound hPF4–heparin to capture HIT antibody, Amiral et al (1996b) failed to detect anti‐hPF4–heparin IgG from 12 HIT sera ( n = 38).…”

Section: Discussionmentioning

confidence: 99%

Anti‐PF4–heparin immunoglobulin G is the major class of heparin‐induced thrombocytopenia antibody: findings of an enzyme‐linked immunofiltration assay using membrane‐bound hPF4–heparin

2001

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An enzyme‐linked immunofiltration assay (ELIFA) was developed for detecting anti‐human platelet factor 4 (hPF4)–heparin antibody in sera of patients with heparin‐induced thrombocytopenia (HIT). The immunofiltration assay was developed to capture HIT antibody by hPF4–heparin complex adsorbed onto a positively charged nylon membrane, as an alternative to plastic bound hPF4. Of 75 sera with a positive serotonin‐release assay (SRA), anti‐PF4–heparin of the immunoglobulin (Ig)G class was detected in 72 (96%) sera. With SRA‐negative sera from thrombocytopenic patients treated with heparin, anti‐hPF4–heparin IgG and IgA were detected in 16% (n = 126) and 14% (n = 74) of sera respectively; 6% (n = 71) of SRA‐negative sera contained both IgG and IgA anti‐hPF4–heparin antibodies. The detection of anti‐hPF4–heparin IgG in all HIT sera supports the assay of anti‐PF4–heparin IgG as being a sensitive screening test for HIT. Alternatively, the absence of anti‐hPF4–heparin IgA cannot be used as a test for excluding HIT, as it was detected in only 48% of SRA‐positive HIT sera. However, it may be used to support the diagnosis of HIT, when HIT IgG is weak. This study emphasized the need to use different immobilizing media for the capture of anti‐PF4–heparin antibody.

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“…The assay was performed under previously reported experimental conditions. 8 14 C-labeled SRA. The biologic effect of the patient's heparin-induced IgG antibody was determined by using 14 Clabeled SRA.…”

Section: Methodsmentioning

confidence: 99%

“…5 Antigen assays determine antibodies toward the PF4/heparin complex by ELISA, 6 heparin-induced IgG by biotin-labeled PF4, 7 and fluorescence-labeled heparin. 8,9 The sensitivity of all assays ranges from 80% to 98%. Antigenic and biologic assays have been compared in several studies to define the sensitivity and specificity.…”

mentioning

confidence: 99%

Laboratory diagnosis of heparin-induced thrombocytopenia type II after clearance of platelet factor 4/heparin complex

Harenberg

Wang

Hoffmann

et al. 2001

Journal of Laboratory and Clinical Medicine

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Determination of heparin‐induced IgG antibody in heparin‐induced thrombocytopenia type II

Cited by 13 publications

References 22 publications

Determination of serotonin release from platelets by enzyme immunoassay in the diagnosis of heparin‐induced thrombocytopenia

Determination of serotonin release from platelets by enzyme immunoassay in the diagnosis of heparin‐induced thrombocytopenia

Anti‐PF4–heparin immunoglobulin G is the major class of heparin‐induced thrombocytopenia antibody: findings of an enzyme‐linked immunofiltration assay using membrane‐bound hPF4–heparin

Laboratory diagnosis of heparin-induced thrombocytopenia type II after clearance of platelet factor 4/heparin complex

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