High Sensitive ELISA Kit for N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP)

NT-Pro-BNP; ; N-BNP

Specificity

This assay has high sensitivity and excellent specificity for detection of High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP).
No significant cross-reactivity or interference between High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-101 89
EDTA plasma(n=5) 92-101 95
heparin plasma(n=5) 86-93 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of High Sensitive N-Terminal Pro-Brain Natriuretic Peptide (NT-ProBNP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 88-103% 84-98% 86-93% 96-104%
EDTA plasma(n=5) 81-101% 96-103% 84-95% 98-105%
heparin plasma(n=5) 85-95% 95-105% 99-105% 87-94%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

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Magazine Citations
Eur Rev Med Pharmacol Sci. The role of serum n-terminal pro-brain natriuretic peptide in transient tachypnea of the newborn Europeanreview: Source
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Integrative Medicine Insights A Randomized Controlled Trial to Study the Effect of Yoga Therapy on Cardiac Function and N Terminal Pro BNP in Heart Failure NCBI: PMC3981569
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Eur J Heart Fail. Growth-differentiation factor 15 and osteoprotegerin in acute myocardial infarction complicated by cardiogenic shock: a biomarker substudy of the IABP-SHOCK II-trial. Pubmed:24903195
Scientific Reports Biomarkers and Echocardiographic Predictors of Myocardial Dysfunction in Patients with Hypertension PubMed: 25747153
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IJC Heart & Vasculature Cardiovascular changes in patients with non-severe Plasmodium vivax malaria article:S2352-9067(16)30010-0
Heart Lung Circ Soluble Glycoprotein 130 and Heat Shock Protein 27 as Novel Candidate Biomarkers of Chronic Heart Failure with Preserved Ejection Fraction Pubmed:27067668
International journal of cardiology Differences in biochemical and genetic biomarkers in patients with heart failure of various etiologies. pubmed:27448535
Experimental and Therapeutic Medicine Expression of pericardial fluid T-cells and related inflammatory cytokines in patients with chronic heart failure. pubmed:28565777
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International Journal of Cardiology 5-methoxytryptophan is a potential marker for post-myocardial infarction heart failure - apreliminary approach to clinical utility. pubmed:27526355
Cardiovasc Toxicol Beneficial Effects of Ozone Therapy on Oxidative Stress, Cardiac Functions and Clinical Findings in Patients with Heart Failure Reduced Ejection Fraction pubmed:28097518
Sci Rep.  Serial heart rhythm complexity changes in patients with anterior wall ST segment elevation myocardial infarction. pubmed:28252107
Journal of Cardiovascular Magnetic Resonance The predictive capabilities of a novel cardiovascular magnetic resonance derived marker of cardiopulmonary reserve on established prognostic surrogate markers in patients with pulmonary vascular disease: results of a longitudinal pilot study pubmed:28065166
Repositório Institucional UNESP AVALIAÇÃO ELETROCARDIOGRÁFICA AMBULATORIAL DE CÃES COM EHRLIQUIOSE MONOCÍTICA CRÔNICA handle:11449
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Original article Evaluation of N-Terminal Pro Brain Natriuretic Peptide as a biomarker for clinical severity of heart failure in pediatric population Attach432
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