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American Diagnostica TAT ELISA product no. 803

www.americandiagnostica.de
A presentation of American Diagnostica/Sekisui's ELISA for quantitative determination of Thrombin/Antithrombin III (TAT) complexes in human plasma samples.

Transcript:
In this presentation you will learn more about American Diagnostica/Sekisui's product No. 803 the so called TAT-Elisa. This is a quantitative enzyme-linked immunosorbent assay for the determination of Thrombin/Antithrombin III complexes in human plasma. The conversion of prothrombin to thrombin is a key event in thrombus formation. Thrombin is a serine protease that acts on various factors in the coagulation pathway and is a major player in thrombus formation. Antithrombin III (ATIII) is the major inhibitor of thrombin. Inactivation of thrombin by antithrombin III leads to the formation of a covalent inactive complex. Inactivation of thrombin, when being generated, proceeds immediately after formation so that the detection of the complex indicates ongoing thrombin formation. In other words: The "Thrombin-Antithrombin III" complex ("TAT"-complex) can be quantified in peripheral blood as an indicator of thrombin activation.

Elevated plasma levels of TAT were found to be associated with a wide variety of active thrombotic events; in particular during disseminated intravascular coagulation, so called DIC; in cases of deep vein thrombosis (DVT); in sepsis and multiple trauma; during pregnancy complications, so called pre-eclampsia, and in a wide variety of malignancies.

What are the key features of the product no. 803?
It is a 96-well plate microtiter format Elisa. The sample type used for the assay is human plasma, be it citrated or EDTA plasma. The assay range is from 0.12 to 7.0 ng/ml. For the assay antibodies specific for human TAT are being used. Some precision date: the intra-assay variation coefficient is 3.3 % and inter-assay CV 12.9 %.

This shows a typical standard curve of TAT concentration versus absorbance at 450 nm.

Here you see data on a correlation study of our IMUBIND TAT ELISA with a comparator product.

The package contains a detailed package insert and it is important to mention that the IMBUBIND TAT-ELISA is CE-marked.

Order and shipping information:
The Imubind TAT Elisa is available from American Diagnostica's German subsidiary, American Diagnostica GmbH. The contact telephone number is shown here (+49 6157 990899). You may use email to info@amdiag.de or order@amdiag.de.

And if you have more specific questions or need more info please contact Andrea Perlik, our Marketing Manager (EMEA) at a.perlik@amdiag.de. It is also important to know that more marketing material for the Imubind TAT Elisa is available at our distributor website (www.adg-distributors.de).

Some information on publications relating to TAT determination.
There was a publication by Fernandez-Cadenas in 2009 who analyzed the thrombin-antithrombin complex in patients treated with t-PA after ischemic stroke. The conclusion drawn from the data was that "...lower levels of TAT were associated with better recanalisation rates at all timepoints..."

Somewhat older, a seminal publication from 1988 by Hoek at al., the major conclusion here is that "TAT concentrations were increased in 25 of the 41 patients who fulfilled the criteria of disseminated intravascular coagulation ... and in 30 of the 35 patients with deep-vein thrombosis of the leg..."

Often I'm asked about the relation of TAT and D-Dimer. There is one publication by Speiser et al. in 1990 who analyzed D-Dimer and TAT in patients with deep vein thrombosis and analyzed their utility in diagnosis and judgment of anticoagulant treatment effectiveness. A major conclusion here is that the determination of the plasma levels of D-Dimer and TAT may be useful for judging the effect of anticoagulant treatment on thrombotic processes.

Again order and shipping information for the TAT-Elisa; important, that the Elisa is sold via the German subsidiary of American Diagnostica/Sekisui and here we have the contact information:

The Imubind TAT ELISA (product no #803) is available from:

American Diagnostica GmbH
Tel: +49 6157 990899
email: info@amdiag.de or order@amdiag.de

If you have questions or need more info, pls contact:
Andrea Perlik (Marketing Manager)
a.perlik@amdiag.de

More marketing material for the Imubind TAT ELISA is available at our distributor website:
www.adg-distributors.de

So, many thanks for your attention!

Prof. Michael Kramer, MD
Medical consultant to
American Diagnostica

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ADI Product 840: APC-R clotting assay

www.american-diagnostica.de
Product presentation ACTICLOT(R) Activated Protein C Resistance (no. 840) of American Diagnostica/Sekisui
Hello, my name is Michael Kramer, I'm medical consultant to American Diagnostica. I want to present a few aspects of American Diagnostica's ACTICLOT Protein C Resistance Assay, Product No. 840, a clotting assay with high specificity and sensitivity for the determination of resistance to activated Protein C.

Activated Protein C Resistance (APC-R) is the most frequent genetic defect associated with deep vein thrombosis. Heterozygous carriers have a 3-7 fold increased risk, homozygous carriers have a 80-100 fold increased risk. About 95% of the APC-R phenotype is caused by the Factor V Leiden mutation, abbreviated as FV:Q506.

Here is a schematic description of the assay principle. The assay contains two snake venoms. 1st the venom RVV-V, a Factor V activating venom from Russel's viper and 2nd Noscarin, a Prothrombin activating venom from the Tiger snake. Noscarin activates Prothrombin to Thrombin, which then converts Fibrinogen to Fibrin and leads to the formation of a clot. Clotting is the analytical endpoint of the assay. The Noscarin mediated Prothrombin to Thrombin activation is accelerated by activated Factor Va, which is generated by the RVV-V venom. Activated Protein C is an inhibitor of Factor Va and once activated Protein C is present and Factor Va is susceptible to inhibition by activated Protein C, this leads to a reduced Noscarin mediated Prothrombin to Thrombin activation, to a reduced Fibrin generation and in the clotting assay to a prolonged clotting-time.
Summary: Activated Factor Va is specifically inhibited by Activated Protein C.
What happens when Factor V is mutated, contains the Factor V Leiden mutation. This leads to resistance of Factor Va to inhibition by activated Protein C. Activated Protein C can no longer inhibit Factor Va. The Factor Va induced acceleration of Prothrombin to Thrombin conversion and Fibrin generation goes on uninhibited. Thus clotting is massively enhanced, the clotting time is reduced.
Summary: Factor V Leiden mutation leads to resistance of activated Factor Va to inactivation by Protein C.

The set-up of the assay and the calculation of results. Basically clotting-time ratios are calculated: a comparison of the clotting time in the presence of activated Protein C versus the clotting-time in the absence of Protein C. The set-up is: plasma plus "840 test reagent" plus APC; and plasma plus test reagent without APC. There are control plasmas to be tested: either negative, which are normal plasmas. Positive plasmas are mostly heterozygous Factor V Leiden mutation carriers. Then ratio calculation, with respect to clotting times, is done.

Here you see a graph of patient samples versus normals, i.e. Wildtype. With respect to Factor V Leiden mutation you see a significant ratio, which means that activated Protein C does inhibit Factor Va in the testing-system. In the case of heterozygous or homozygous genotypes, you see a significant reduction of the ratio. There is a reasonable and significant difference between wildtype and heterozygous genotypes.
Summary: ACTICLOT APC-R Test clearly distinguishes between Wildtype, Homozygous and Heterozygous Factor V Leiden (FV:Q506) Mutation Carriers.

What are the mayor advantages of the assay?
1st a clear discrimination between wildtype, heterozygous and homozygous individuals. 2nd a sensitivity very similar to Polymerase-Chain-Reaction (PCR). In contrast to PCR-technology, the clotting test is cost and time saving. The testing can be done with standard clotting instrumentation. The assay is not influenced by a variety of coagulation abnormalities (Lupus anticoagulant, FVIII, abnormal PT, aPTT, Protein C or Protein S, heparins, different FIIa activities, fibrinogens, D-Dimer or antithrombin plasma levels). Protocols are available for all major routine analyzers.

The package-content of Product 840 allows 120 determinations (3 x 40). Control plasmas are available, negative control plasmas from individuals which do not contain the Factor V Leiden Mutation and positive control plasmas, from individuals that are genetically heterozygous for the Factor V Leiden Mutation. The assay is CE-Marked.

The APC-R Kit and the Control Plasmas are available from the German subsidiary of American Diagnostica, American Diagnostica GmbH. Please contact Dr. Martina Kloeppinger, the Managing Director. Send an email to info@amdiag.de or order@amdiag.de. Instrument applications for the ACTICLOT Protein C Resistance Assays and protocols for all mayor routine analyzers are available. New Instrument Applications are added from time to time.

Additional marketing material that can be used for marketing of Product 840 is available at American Diagnostica GmbH's distributor website www.adg-distributors.de.

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