A quantitative detection of Cardiotrophin-1 in chronic heart failure by chemiluminescence immunoassay
Ying Ping1, Xuchu Wang1, Yibei Dai1
1Department of Laboratory Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.
Insights
A new assay accurately detects cardiotrophin-1 (CT-1) in plasma, a potential biomarker for heart failure. This rapid, automated method offers high sensitivity and reproducibility for clinical use.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Immunology
Background:
- Cardiotrophin-1 (CT-1) is a cytokine linked to cardiomyocyte hypertrophy and dysfunction.
- Elevated plasma CT-1 levels may indicate heart failure, useful for diagnosis, staging, and prognosis.
Purpose of the Study:
- To establish a rapid and sensitive method for detecting plasma CT-1.
- To evaluate the clinical applicability of the developed assay for heart failure assessment.
Main Methods:
- A one-step paramagnetic particles-based chemiluminescence immunoassay (MPs-CILA) was developed.
- The assay utilizes biotin-labeled and acridine ester-labeled anti-CT-1 antibodies for sandwich complex formation.
- Paramagnetic particles facilitate rapid separation and signal generation.
Main Results:
- The MPs-CILA demonstrated a wide linear range (7.8 pg/mL to 200 ng/mL) with a low detection limit (1.0 pg/mL).
- High recovery rates (96-110%) and acceptable intra- and inter-assay coefficients of variation (CVs) confirmed precision and reproducibility.
- The assay was applied to analyze plasma CT-1 in 140 subjects with and without chronic heart failure.
Conclusions:
- The developed MPs-CILA is highly sensitive, precise, and reproducible for plasma CT-1 detection.
- The assay's automation makes it suitable for high-throughput clinical screening.
- This method can aid in the diagnosis and management of heart failure.
Background:
Cardiotrophin-1 (CT-1) is a cytokine that could induce cardiomyocytes hypertrophy and dysfunction. Plasma CT-1 might serve as a cardiac biomarker both in diagnosis, staging, and prognostic assessment of heart failure.
Methods:
In this study, a one-step paramagnetic particles-based chemiluminescence immunoassay (MPs-CILA) for rapid and sensitive detection of plasma CT-1 was established. Plasma samples were directly incubated with biotin-labeled anti-CT-1 antibody (bio-Ab) and acridine ester labeled anti-CT-1 antibody (AE-Ab) to form sandwiched complex. The sandwiched CT-1 was then captured by streptavidin modified paramagnetic particles (MPs-SA) for rapid separation and signal generation.
Results:
The proposed MPs-CLIA presents a laudable linear relationship ranging from 7.8 pg/mL to 200 ng/mL with a detection limit of 1.0 pg/mL. The recoveries of spiked human plasma samples at low (10pg/mL), medium (100 pg/mL), and high (800 pg/mL) levels of CT-1 were 96%, 104%, and 110% respectively. The intra-analysis coefficient variation (CVs) of the 3 samples was 8.92%, 6.69%, and 3.54%, respectively. And the inter-analysis coefficient variation (CVs) was 9.25%, 10.9%, and 4.3%, respectively. These results strongly indicate high sensitivity, wide linear range, acceptable precision, and applicable reproducibility of the proposed method to detect plasma level of CT-1. Finally, Plasma CT-1 from 140 subjects with or without chronic heart failure was analyzed to assess the clinical application of MPs-CILA.
Conclusions:
Noteworthily, the MPs-CLIA method is highly automated such that it is suitable for high-throughput detection of CT-1 in clinical inspection.
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