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Filter Paper Blood Spot Enzyme Linked Immunoassay for Adiponectin and Application in the Evaluation of Determinants of Child Insulin Sensitivity
- Source :
- PLoS ONE, Vol 8, Iss 8, p e71315 (2013), PLoS ONE
- Publication Year :
- 2013
- Publisher :
- Public Library of Science (PLoS), 2013.
-
Abstract
- Background Adiponectin is an adipocyte-derived hormone that acts as a marker of insulin sensitivity. Bloodspot sampling by fingerstick onto filter paper may increase the feasibility of large-scale studies of the determinants of insulin sensitivity. We first describe the validation of an enzyme-linked immunoassay (ELISA) for quantifying adiponectin from dried blood spots and then demonstrate its application in a large trial (PROBIT). Methods We quantified adiponectin from 3-mm diameter discs (≈3 µL of blood) punched from dried blood spots obtained from: i) whole blood standards (validation); and ii) PROBIT trial samples (application) in which paediatricians collected blood spots from 13,879 children aged 11.5 years from 31 sites across Belarus. We examined the distribution of bloodspot adiponectin by demographic and anthropometric factors, fasting insulin and glucose. Results In the validation study, mean intra-assay coefficients of variation (n = 162) were 15%, 13% and 10% for ‘low’ (6.78 µg/ml), ‘medium’ (18.18 µg/ml) and 'high’ (33.13 µg/ml) internal quality control (IQC) samples, respectively; the respective inter-assay values (n = 40) were 23%, 21% and 14%. The correlation coefficient between 50 paired whole bloodspot versus plasma samples, collected simultaneously, was 0.87 (95% CI: 0.78 to 0.93). Recovery of known quantities of adiponectin (between 4.5 to 36 µg/ml) was 100.3–133%. Bloodspot adiponectin was stable for at least 30 months at −80°C. In PROBIT, we successfully quantified fasting adiponectin from dried blood spots in 13,329 of 13,879 (96%) children. Mean adiponectin (standard deviation) concentrations were 17.34 µg/ml (7.54) in boys and 18.41 µg/ml (7.92) in girls and were inversely associated with body mass index, fat mass, triceps and subscapular skin-fold thickness, waist circumference, height and fasting glucose. Conclusions Bloodspot ELISA is suitable for measuring adiponectin in very small volumes of blood collected on filter paper and can be applied to large-scale studies.
- Subjects :
- Male
Anatomy and Physiology
Epidemiology
medicine.medical_treatment
lcsh:Medicine
030204 cardiovascular system & hematology
Endocrinology
0302 clinical medicine
Pediatric Endocrinology
Blood plasma
Pathology
Insulin
Medicine
030212 general & internal medicine
Child
lcsh:Science
2. Zero hunger
Multidisciplinary
medicine.diagnostic_test
Child Health
Clinical Laboratory Sciences
3. Good health
Female
Public Health
Adiponectin
Research Article
Test Evaluation
Adult
Paper
medicine.medical_specialty
Adolescent
Fingerstick
Endocrine System
Enzyme-Linked Immunosorbent Assay
03 medical and health sciences
Insulin resistance
Diagnostic Medicine
Internal medicine
Humans
Diabetic Endocrinology
Endocrine Physiology
Filter paper
business.industry
lcsh:R
Reproducibility of Results
Insulin sensitivity
medicine.disease
Immunoassay
Immunology
lcsh:Q
Dried Blood Spot Testing
Insulin Resistance
business
Biomarkers
Filtration
General Pathology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 8
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....8dcc12ad2315a3e8c78f272c732ee158