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Differences in Cardiac Troponin T Composition in Myocardial Infarction and End-Stage Renal Disease Patients: A Blood
Wim H M Vroemen1, Ellen J S Denessen1,2, William P T M van Doorn1
1Central Diagnostic Laboratory, Maastricht University Medical Center, Maastricht, the Netherlands.
Insights
Cardiac troponin T (cTnT) degradation varies by anticoagulation method, impacting proteoform composition but not concentration. Larger cTnT proteoforms indicate myocardial infarction (MI), while smaller fragments suggest end-stage renal disease (ESRD).
Area of Science:
- Biochemistry
- Clinical Chemistry
- Cardiovascular Diagnostics
Background:
- Cardiac troponin T (cTnT) is crucial for diagnosing myocardial infarction (MI).
- Elevated cTnT levels in end-stage renal disease (ESRD) patients complicate diagnosis.
- Previous studies suggested pre-analytic effects, like thrombin generation in serum, influence cTnT findings.
Purpose of the Study:
- To investigate the impact of different anticoagulation methods on cTnT concentration and proteoform composition.
- To compare cTnT proteoform profiles between MI and ESRD patients.
Main Methods:
- Simultaneous collection of serum, lithium-heparin (LH) plasma, and ethylenediaminetetraacetic acid (EDTA) plasma.
- Analysis using a high-sensitivity (hs-)cTnT immunoassay.
- Investigation of cTnT proteoform composition via spike experiments and gel filtration chromatography (GFC).
Main Results:
- High agreement in hs-cTnT concentrations across serum, LH plasma, and EDTA plasma.
- Significant time-dependent degradation of cTnT observed in LH and EDTA plasma, and serum, with distinct proteoform ratios.
- MI patients predominantly exhibited larger cTnT proteoforms, whereas ESRD patients showed mainly small 15-18 kDa fragments.
Conclusions:
- In vitro cTnT degradation is influenced by the anticoagulation method but does not affect hs-cTnT concentrations.
- Distinct cTnT proteoform profiles differentiate MI (larger proteoforms) from ESRD (small fragments).
- These findings are vital for developing novel hs-cTnT assays targeting specific proteoforms.
Background:
Cardiac troponin T (cTnT) is key in diagnosing myocardial infarction (MI) but is also elevated in end-stage renal disease (ESRD) patients. Specific larger cTnT proteoforms were identified for the acute phase of MI, while in serum of ESRD patients solely small cTnT fragments were found. However, others allocated this to a pre-analytic effect due to abundant thrombin generation in serum. Therefore, we investigated the effect of various anticoagulation methods on cTnT composition and concentration and compared the cTnT composition of MI and ESRD patients.
Methods:
The agreement of cTnT concentrations between simultaneously collected serum, lithium-heparin (LH) plasma, and ethylenediaminetetraacetic acid (EDTA) plasma was studied using the high-sensitivity (hs-)cTnT immunoassay. cTnT proteoform composition was investigated in a standardized time-dependent manner through spike experiments and in simultaneously collected blood matrixes of MI and ESRD patients.
Results:
Excellent hs-cTnT concentration agreements were observed across all blood matrixes (slopes > 0.98; 95% CI, 0.96-1.04). Time-dependent degradation (40 kDa intact:29 kDa fragment:15 to 18 kDa fragments) was found in LH plasma and EDTA plasma, and serum in ratios (%) of 90:10:0, 0:5:95, and 0:0:100, respectively (48 h after blood collection). Moreover, gel filtration chromatography (GFC) profiles illustrated mainly larger cTnT proteoforms in MI patients, while in ESRD patients mainly 15 to 18 kDa fragments were found for all matrices.
Conclusions:
The extent of cTnT degradation in vitro is dependent on the (anti)coagulation method, without impacting hs-cTnT concentrations. Furthermore, mainly larger cTnT proteoforms were present in MI patients, while in ESRD patients mainly small 15 to 18 kDa cTnT fragments were found. These insights are essential when developing a novel hs-cTnT assay targeting larger cTnT proteoforms.
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