A Lysis- and Sonication-Based Method for the Quantification of Extracellular Vesicle-Bound Cardiac Troponin T Using a High-Sensitivity Immunoassay

Q: What are the key findings of this study?

A combined lysis and sonication protocol significantly increased measurable cTnT levels by 10% to 34% compared to native samples by releasing EV-bound cTnT, reducing processing time to 10 minutes.

Q: What is the clinical evidence from this study?

Study design: Observational. Population: Myocardial injury (n=98). Intervention: Combined lysis and sonication protocol vs. Native, unprocessed plasma and conventional EV isolation. Primary outcome: hs-cTnT levels compared to native, unprocessed samples (p=<0.001).

Q: What does this research mean for the field?

A rapid lysis- and sonication-based protocol liberates extracellular vesicle-bound cardiac troponin T in approximately 10 minutes, significantly increasing measurable cTnT concentrations by 10% to 34% across cardiovascular disease cohorts and healthy controls. Novelty: ClaimNovelty.METHODOLOGICAL. Consensus alignment: ConsensusAlignment.NEUTRAL.

Q: What question did this study set out to answer?

To establish and validate a rapid lysis- and sonication-based protocol that releases extracellular vesicle-bound cardiac troponin T for quantification via standard high-sensitivity immunoassay.

Y. Ding; Dominika Bernáth-Nagy; Chiara Heß; Florian Leuschner; Hugo A. Katus; Norbert Frey; Jona B. Krohn; Evangelos Giannitsis

doi:10.3390/biomedicines14081653

July 23, 2026BiomedicinesOpen Access

A Lysis- and Sonication-Based Method for the Quantification of Extracellular Vesicle-Bound Cardiac Troponin T Using a High-Sensitivity Immunoassay

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Key result

Rapid lysis-sonication protocol increases measurable cTnT up to ~34% by releasing extracellular vesicle-bound troponin.

P<0.001
n=98

Figures

Why the study?

A fraction of circulating cTnT enclosed within extracellular vesicles goes widely undetected by standard hs-cTn assays.

Does a combined lysis and sonication protocol improve the rapid quantification of EV-bound cTnT compared to native plasma and conventional EV isolation in patients with various cardiovascular conditions?

Key Points

To establish and validate a rapid lysis- and sonication-based protocol that releases extracellular vesicle-bound cardiac troponin T for quantification via standard high-sensitivity immunoassay.
Treated plasma with a lysis buffer containing 0.1% Triton X-100 and protease/phosphatase inhibitor followed by low-intensity sonication (5 cycles of 30 s ON / 30 s OFF at 4 °C) to lyse extracellular vesicles.
Quantified cardiac troponin T with a fifth-generation high-sensitivity immunoassay on Cobas analyzers and compared performance against conventional exosome precipitation.
Combined lysis and sonication significantly increased detectable cardiac troponin T over unprocessed samples in all cohorts, with median increases ranging from ~10% in decompensated aortic stenosis to ~34% in young healthy controls.
Extracellular vesicle-bound cardiac troponin T accounted for ~15% of total circulating plasma troponin T in NSTEMI patients and remained stable across 72 h.
Pre-analytical turnaround time dropped from ~2.5 h using conventional precipitation isolation to ~10 min with the combined lysis and sonication method.

Study Design

Type

Observational (n=98)

Multicenter

No

Structured PICO

Does a combined lysis and sonication protocol improve the rapid quantification of EV-bound cTnT compared to native plasma and conventional EV isolation in patients with various cardiovascular conditions?

P

Population

98 participants, including patients with various cardiovascular diseases such as NSTEMI and healthy controls, whose blood samples were analyzed to validate a novel method for quantifying EV-bound cardiac troponin T.

I

Intervention

Combined lysis (0.1% Triton X-100) and low-intensity sonication protocol prior to high-sensitivity cardiac troponin T (hs-cTnT) immunoassay.

C

Comparator

Native, unprocessed plasma samples and a conventional kit-based extracellular vesicle (EV) isolation method.

O

Outcome

Total cTnT post EV disruption (quantification of EV-bound cTnT).surrogate

Main Result

p-value: p=<0.001

Limitations

Healthy controls in the primary study cohort were not age-matched to the patient cohorts, potentially confounding results with age-related differences in EV biology.
Long-term storage stability experiments were exploratory and require further confirmation in future studies.

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Implication

A novel 10-minute lysis and sonication protocol successfully liberates previously undetected EV-bound cardiac troponin T, enabling its rapid quantification using standard high-sensitivity immunoassays in clinical routine.

Abstract

Background/Objectives: High-sensitivity cardiac troponin (hs-cTn) assays are used in routine diagnostics to detect myocardial injury. However, a fraction of circulating cardiac troponin T (cTnT) enclosed within extracellular vesicles (EVs) goes widely undetected. This study introduces a combined lysis- and sonication-based protocol to release and quantify EV-bound cTnT in a time-efficient manner using a state-of-the-art hs-cTnT immunoassay. Methods: Plasma samples from patients with non-ST-segment elevation myocardial infarction (NSTEMI), unstable angina, pulmonary embolism, decompensated aortic stenosis, atrial fibrillation, myocarditis, and healthy controls were treated with a lysis buffer and subsequently sonicated. Treated and untreated samples were assessed and compared to a conventional EV isolation method. Results: Following combined lysis and sonication, cTnT levels were significantly higher compared to native, unprocessed samples across all cohorts. The median increase post-processing ranged from ~10% in decompensated aortic stenosis to ~34% in young healthy controls. In NSTEMI, the EV-bound cTnT accounted for ~15% of plasma cTnT and remained stable over 72 h. The EV cTnT/plasma cTnT ratios were comparable between the combined lysis and sonication approach and the EV isolation method. Processing time prior to cTnT measurement was reduced from ~2.5 h to ~10 min using the lysis and sonication protocol. Conclusions: Our method allows for the rapid liberation of a previously inaccessible EV-bound fraction of cTnT without the need for time-consuming and resource-intensive EV isolation workflows and is therefore readily implementable alongside standard hs-cTnT testing. The observed EV-cTnT patterns suggest differential compartmentation of cTnT, potentially reflecting the myocardial pathophysiology underlying troponin elevation.

Discussion

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Cite This Study

Ding et al. (2026) conducted an observational in Myocardial injury (n=98). Combined lysis and sonication protocol vs. Native, unprocessed plasma and conventional EV isolation was evaluated on hs-cTnT levels compared to native, unprocessed samples (p=<0.001). A combined lysis and sonication protocol significantly increased measurable cTnT levels by 10% to 34% compared to native samples by releasing EV-bound cTnT, reducing processing time to 10 minutes.

synapsesocial.com/papers/6a65a6b5d3aea3239cd77d65 https://doi.org/10.3390/biomedicines14081653

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A Lysis- and Sonication-Based Method for the Quantification of Extracellular Vesicle-Bound Cardiac Troponin T Using a High-Sensitivity Immunoassay

Figures

Key Points

Study Design

Structured PICO

Main Result

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

A Lysis- and Sonication-Based Method for the Quantification of Extracellular Vesicle-Bound Cardiac Troponin T Using a High-Sensitivity Immunoassay

Figures

Key Points

Study Design

Structured PICO

Main Result

Limitations

Ask about this paper

Implication

Abstract

Discussion

Cite This Study

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