Fully Automated Ultrasensitive Digital Immunoassay for Cardiac Troponin I Based on Single Molecule Array Technology

Jarolím, Petr; Patel, Purvish P.; Conrad, Michael; Chang, Lei; Melenovský, Vojtěch; Wilson, David H.

doi:10.1373/clinchem.2015.242081

Cited by 22 publications

(23 citation statements)

References 30 publications

(27 reference statements)

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“…47 Disadvantages of the method are that it requires relatively large sample volumes, up to 100 uL, and is limited to single proteins. 23 Another method referred to as digital ELISA, offered by Quanterix (Lexington, MA), has also been shown to detect troponin in this lower range 48 and is based on analyte enrichment with microbeads which are each loaded into a femtoliter microwell, avoiding antibody cross-reactions, and converted to fluorescence products using enzymatic methods. 49 Digital ELISA has been multiplexed to measure six low-abundance cytokines in parallel.…”

Section: Increasing the Sensitivity Of Immunoaffinity Assays To Singlmentioning

confidence: 99%

Emerging Affinity-Based Proteomic Technologies for Large-Scale Plasma Profiling in Cardiovascular Disease

Smith¹,

2017

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Plasma biomarkers that reflect molecular states of the cardiovascular system are central for clinical decision-making. Routinely used plasma biomarkers include troponins, natriuretic peptides and lipoprotein particles, yet interrogate only a modest subset of pathways relevant to cardiovascular disease. Systematic profiling of a larger portion of circulating plasma proteins (the plasma proteome) will provide opportunities for unbiased discovery of novel markers to improve diagnostic or predictive accuracy. In addition, proteomic profiling may inform pathophysiological understanding and point to novel therapeutic targets. Obstacles for comprehensive proteomic profiling include the immense size and structural heterogeneity of the proteome, as well as the broad range of abundance levels. Proteome-wide, untargeted profiling can be performed in tissues and cells with tandem mass spectrometry. However, applications to plasma are limited by the need for complex preanalytical sample preparation stages limiting sample throughput. Multiplexing of targeted methods based on capture and detection of specific proteins are therefore receiving increasing attention in plasma proteomics. Immunoaffinity assays are the workhorse for measuring individual proteins but have been limited for proteomic applications by long development times, cross-reactivity preventing multiplexing, specificity issues, and incomplete sensitivity to detect proteins in the lower range of the abundance spectrum (below picograms per milliliter). Emerging technologies to address these issues include nucleotide-labeled immunoassays and aptamer reagents which can be automated for efficient multiplexing of thousands of proteins at high sample throughput, coupling of affinity capture methods to mass spectrometry for improved specificity, and ultrasensitive detection systems to measure low-abundance proteins. In addition, proteomics can now be integrated with modern genomics tools to comprehensively relate proteomic profiles to genetic variants, which may both influence binding of affinity reagents and serve to validate the target specificity of affinity assays. The application of deep quantitative proteomic profiling to large cohorts has thus become increasingly feasible with emerging affinity methods. The aims of this article are to provide the broad readership of Circulation with a timely overview of emerging methods for affinity proteomics and recent progress in cardiovascular medicine based on such methods.

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Section: Increasing the Sensitivity Of Immunoaffinity Assays To Singlmentioning

confidence: 99%

Emerging Affinity-Based Proteomic Technologies for Large-Scale Plasma Profiling in Cardiovascular Disease

Smith¹,

2017

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“…In a research the level of cTnI was significantly higher in heart failure patients, and exhibited increasing precise concentrations with rising New York Heart Association classification of heart failure severity, which meant cTnI is an accurate indicator for the diagnosis of heart failure [32]. There are also other studies demonstrated that the release of cTnI is associated with increased 1 year mortality and heart failure related readmission, and the cTnI level can act as an independent predicator, and complete in prognostic utility of CHF patients [33].…”

Section: Discussionmentioning

confidence: 99%

Effects of Active Components of Fuzi and Gancao Compatibility on Bax, Bcl‐2, and Caspase‐3 in Chronic Heart Failure Rats

Wang

Zhang

Zhou

et al. 2016

Evidence-Based Complementary and Alternative Medicine

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Hypaconitine (HA) and glycyrrhetinic acid (GA) are active components of Fuzi (Aconitum carmichaelii) and Gancao (Glycyrrhiza uralensis Fisch); they have been used in compatibility for chronic heart failure (CHF) from ancient times. The purpose of the present research was to explore whether apoptosis pathways were related with the protective effects of HA + GA against CHF rats or not. The rats were progressed with transverse-aortic constriction (TAC) operation for 4 weeks to build the CHF state, and then the Digoxin (1 mg/kg), HA (2.07 mg/kg), GA (25 mg/kg), and HA (2.07 mg/kg) + GA (25 mg/kg) were orally administrated to rats for 1 week. The levels of BNP and cTnI in the plasma were decreased in the HA + GA group, and the heart/body weight ratio (H/B) and left ventricular (LV) parameters of transthoracic echocardiography were also declined; moreover, the expressions of Bax, Bcl-2, and caspase-3 were all improved in the HA + GA group than other groups in the immunohistochemistry and western blot methods. In general, the data suggested that Fuzi and Gancao compatibility could protect the CHF rats from apoptosis, which provided a strong evidence for further searching for mechanisms of them.

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“…The sandwich-type immunosensor format allocates 40 min to allow for cTnI binding and an additional 80 min to accommodate the binding of the IRMOF-based detection probe. An alternative high sensitivity fluorescence-based detection assay capitalises on Single-molecule Array (Simoa) technology [123]. Simoa is based on “the simultaneous counting of singulated capture microbeads” to detect the target analyte [124].…”

Section: Evaluating Ctni Detection Techniquesmentioning

confidence: 99%

“…Figure 3 and Figure 4: [37,38,48,49,50,57,58,61,65,66,68,69,70,71,74,84,85,86,87,88,89,93,94,95,96,97,104,105,111,112,115,116,117,120,121,122,123,125,126,127,128,130,132,133,134,136,138,144,145,148,150,151,159,161,174,175,176,184,185,186,187,188,189,190,…”

Section: Reprinting Figuresmentioning

confidence: 99%

Point-of-Care Compatibility of Ultra-Sensitive Detection Techniques for the Cardiac Biomarker Troponin I—Challenges and Potential Value

2018

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Cardiac biomarkers are frequently measured to provide guidance on the well-being of a patient in relation to cardiac health with many assays having been developed and widely utilised in clinical assessment. Effectively treating and managing cardiovascular disease (CVD) relies on swiftly responding to signs of cardiac symptoms, thus providing a basis for enhanced patient management and an overall better health outcome. Ultra-sensitive cardiac biomarker detection techniques play a pivotal role in improving the diagnostic capacity of an assay and thus enabling a better-informed decision. However, currently, the typical approach taken within healthcare depends on centralised laboratories performing analysis of cardiac biomarkers, thus restricting the roll-out of rapid diagnostics. Point-of-care testing (POCT) involves conducting the diagnostic test in the presence of the patient, with a short turnaround time, requiring small sample volumes without compromising the sensitivity of the assay. This technology is ideal for combatting CVD, thus the formulation of ultra-sensitive assays and the design of biosensors will be critically evaluated, focusing on the feasibility of these techniques for point-of-care (POC) integration. Moreover, there are several key factors, which in combination, contribute to the development of ultra-sensitive techniques, namely the incorporation of nanomaterials for sensitivity enhancement and manipulation of labelling methods. This review will explore the latest developments in cardiac biomarker detection, primarily focusing on the detection of cardiac troponin I (cTnI). Highly sensitive detection of cTnI is of paramount importance regarding the rapid rule-in/rule-out of acute myocardial infarction (AMI). Thus the challenges encountered during cTnI measurements are outlined in detail to assist in demonstrating the drawbacks of current commercial assays and the obstructions to standardisation. Furthermore, the added benefits of introducing multi-biomarker panels are reviewed, several key biomarkers are evaluated and the analytical benefits provided by multimarkers-based methods are highlighted.

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Fully Automated Ultrasensitive Digital Immunoassay for Cardiac Troponin I Based on Single Molecule Array Technology

Cited by 22 publications

References 30 publications

Emerging Affinity-Based Proteomic Technologies for Large-Scale Plasma Profiling in Cardiovascular Disease

Emerging Affinity-Based Proteomic Technologies for Large-Scale Plasma Profiling in Cardiovascular Disease

Effects of Active Components of Fuzi and Gancao Compatibility on Bax, Bcl‐2, and Caspase‐3 in Chronic Heart Failure Rats

Point-of-Care Compatibility of Ultra-Sensitive Detection Techniques for the Cardiac Biomarker Troponin I—Challenges and Potential Value

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