| Background: Heart failure(HF)is the end stage of all heart diseases.Its morbidity and mortality have increased year by year,and it has caused great economic burden on humans.Therefore,it is important to identify patients with chronic heart failure early.Cardiotrophin-1(CT-1),a member of the interleukin-6 superfamily,is a cytokine that could induce cardiomyocytes hypertrophy and dysfunction.Amounts of studies indicated that plasma CT1 might serve as a cardiac biomarker both in diagnosis,staging,and prognostic assessment of heart failure.However,the lack of a rapid and simple technique for the quantitative analysis of CT-1 greatly limits the application of CT-1 in clinical research.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.1.Establishment of the MPs-CILA platform using biotin-labeled rabbit anti-human CT-1 polyclonal antibody(bio-Ab)and acridine ester labeled rabbit anti-human CT-1polyclonal antibody(AE-Ab).Optimization of MPs-CLIA experimental condition,including antibodies concentration,reaction time,capture time and so on.2.Evaluation of MPs-CLIA3.Assess the clinical application of MPs-CILA.Plasma CT-1 from 140 subjects with or without chronic heart failure was analyzed to assess the clinical application of MPs-CILA and Preliminary evaluated the diagnostic value of CT-1 for chronic heart failure.Results:1.The proposed MPs-CLIA presents a laudable linear relationship ranging from7.8pg/m L to 2000pg/m L with a detection limit of 2.6pg/m L.Calibration curve was RLU=32.646CCT1+174.35(R2=0.9996).The recoveries of spiked human plasma samples at low(10pg/m L),medium(100pg/m L)and high(800 pg/m L)levels of CT-1 were 96%,104%,110% respectively.The intra-analysis coefficient variation(CVs)of the 3 samples were 8.92%,6.69% and 3.54%,respectively.And the inter-analysis coefficient variation(CVs)were 9.25%,10.9% and 4.3%respectively.No cross-reaction with IL-6 and BNP.These results strongly indicate high sensitivity,wide linear range,acceptable precision,and applicable reproducibility of the proposed method to detect plasma level of CT-1.2.The CT-1 concentrations were significantly higher in CHF patients(median:70.43pg/m L)compared with healthy individuals(median: 40.70pg/m L)(P<0.05),ROC curve was further performed to evaluate the diagnostic value of CT-1,which yielded an area under the curve of 0.66.Regression analysis revealed significant association between CT-1 and pro-BNP(R2 =0.398,P<0.001).Conclusions: The performance indicators of the MPs-CLIA methodology established in this study meet the clinical needs.Noteworthily,the MPs-CLIA method is highly automated such that it is suitable for high-throughput detection of CT-1 in clinical inspection.And the detection of plasma CT-1 has a good application prospect for the diagnosis of heart failure. |
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