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Automation and Analytical Techniques |
1 Department of Clinical Biochemistry, Statens Serum Institut, Copenhagen, Denmark.
2 NANEA at Department of Epidemiology and Social Medicine, Aarhus University, Aarhus, Denmark.
3 National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention, Atlanta, GA.
4 Department of Pediatrics, Copenhagen University Hospital, Hvidovre, Denmark.
aAddress correspondence to this author at: Department of Clinical Biochemistry, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark. Fax 45-3268-3878; e-mail dh{at}ssi.dk.
Background: Inflammatory reactions and other events in early life may be part of the etiology of late-onset diseases, including cerebral palsy, autism, and type 1 diabetes. Most neonatal screening programs for congenital disorders are based on analysis of dried blood spot samples (DBSS), and stored residual DBSS constitute a valuable resource for research into the etiology of these diseases. The small amount of blood available, however, limits the number of analytes that can be determined by traditional immunoassay methodologies.
Methods: We used new multiplexed sandwich immunoassays based on flowmetric Luminex® xMAP technology to measure inflammatory markers and neutrophins in DBSS.
Results: The high-capacity 25-plex multianalyte method measured 23 inflammatory and trophic cytokines, triggering receptor expressed on myeloid cells-1 (TREM-1), and C-reactive protein in two 3.2-mm punches from DBSS. It also measured 26 cytokines and TREM-1 in serum. Standards Recovery in the 25-plex method were 90%–161% (mean, 105%). The low end of the working range for all 25 analytes covered concentrations found in DBSS from healthy newborns. Mean recovery of exogenous analytes added at physiologic concentrations in DBSS models was 174%, mean intra- and interassay CVs were 6.2% and 16%, respectively, and the mean correlation between added and measured analytes was r2 = 0.91. In DBSS routinely collected on days 5–7 from 8 newborns with documented inflammatory reactions at birth, the method detected significantly changed concentrations of inflammatory cytokines. Measurements on DBSS stored at –24 °C for >20 years showed that most cytokines are detectable in equal concentrations over time.
Conclusions: The method can reliably measure 25 inflammatory markers and growth factors in DBSS. It has a large potential for high-capacity analysis of DBSS in epidemiologic case–control studies and, with further refinements, in neonatal screening.
The following articles in journals at HighWire Press have cited this article:
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  A.-D. Gu, H.-Y. Mo, Y.-B. Xie, R.-J. Peng, J.-X. Bei, J. Peng, M.-Y. Li, L.-Z. Chen, Q.-S. Feng, W.-H. Jia, et al. Evaluation of a Multianalyte Profiling Assay and an Enzyme-Linked Immunosorbent Assay for Serological Examination of Epstein-Barr Virus-Specific Antibody Responses in Diagnosis of Nasopharyngeal Carcinoma Clin. Vaccine Immunol., November 1, 2008; 15(11): 1684 - 1688. [Abstract] [Full Text] [PDF]
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 A.-D. Gu, Y.-B. Xie, H.-Y. Mo, W.-H. Jia, M.-Y. Li, M. Li, L.-Z. Chen, Q.-S. Feng, Q. Liu, C.-N. Qian, et al. Antibodies against Epstein-Barr virus gp78 antigen: a novel marker for serological diagnosis of nasopharyngeal carcinoma detected by xMAP technology J. Gen. Virol., May 1, 2008; 89(5): 1152 - 1158. [Abstract] [Full Text] [PDF]
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 K. Skogstrand, D. M. Hougaard, D. E. Schendel, N.-P. Bent, C. Svaerke, and P. Thorsen Association of Preterm Birth With Sustained Postnatal Inflammatory Response Obstet. Gynecol., May 1, 2008; 111(5): 1118 - 1128. [Abstract] [Full Text] [PDF]
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 E. L Madsen, A. Rissanen, J. M Bruun, K. Skogstrand, S. Tonstad, D. M Hougaard, and B. Richelsen Weight loss larger than 10% is needed for general improvement of levels of circulating adiponectin and markers of inflammation in obese subjects: a 3-year weight loss study Eur. J. Endocrinol., February 1, 2008; 158(2): 179 - 187. [Abstract] [Full Text] [PDF]
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A Klamer, K Skogstrand, D M Hougaard, B Norgaard-Petersen, A Juul, and G Greisen Adiponectin levels measured in dried blood spot samples from neonates born small and appropriate for gestational age Eur. J. Endocrinol., August 1, 2007; 157(2): 189 - 194. [Abstract] [Full Text] [PDF]
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 M. V. Hollegaard, K. M. Sorensen, H. K. Petersen, M. B. Arnardottir, B. Norgaard-Pedersen, P. Thorsen, and D. M. Hougaard Whole Genome Amplification and Genetic Analysis after Extraction of Proteins from Dried Blood Spots Clin. Chem., June 1, 2007; 53(6): 1161 - 1162. [Full Text] [PDF]
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 I. Vanhorebeek, R. P. Peeters, S. Vander Perre, I. Jans, P. J. Wouters, K. Skogstrand, T. K. Hansen, R. Bouillon, and G. Van den Berghe Cortisol Response to Critical Illness: Effect of Intensive Insulin Therapy J. Clin. Endocrinol. Metab., October 1, 2006; 91(10): 3803 - 3813. [Abstract] [Full Text] [PDF]
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K. Kofoed, U. V. Schneider, T. Scheel, O. Andersen, and J. Eugen-Olsen Development and Validation of a Multiplex Add-On Assay for Sepsis Biomarkers Using xMAP Technology Clin. Chem., July 1, 2006; 52(7): 1284 - 1293. [Abstract] [Full Text] [PDF]
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