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Technical Briefs |
1
Division of Genetic Disorders, Wadsworth Center, New York State Department of Health, Albany, NY 12201-0509
2
Department of Biomedical Sciences, School of Public Health, University at Albany, One University Place, Rensselaer, NY 12144-3456
a author for correspondence: fax 518-473-8627, e-mail bellisar@wadsworth.org
All newborns in the United States are screened by state-sponsored programs for congenital hypothyroidism. Traditionally, this has involved an initial screen for thyroxine (T4), followed by testing thyrotropin (TSH) concentrations of infants with low T4 values. This algorithm or an alternative whereby only TSH is assayed does not detect all hypothyroid newborns (1). A simultaneous assay that measures both T4 and TSH would be preferred for screening. Luminex MultiAnalyte Profiling Technology (LabMAP) was used to develop an immunoassay that measures T4 and TSH simultaneously from a single blood-spot sample. The method uses a mixture of two distinct sets of uniquely fluorescent microspheres, which are identified by distinct red and orange fluorescent dyes by the Luminex100 flow analyzer (2). Quantification is accomplished with a green fluorescent reporter molecule. At present, 100 distinct sets of fluorescent microspheres are available, permitting the simultaneous measurement of as many as 100 different analytes.
Uniquely, LabMAP provides the ability to combine different immunoassay
formats in a single detection system: a competitive-inhibition format
for T4 measurement and a sandwich-capture format
for measurement of TSH. Using two Luminex spectral-addressable
fluorescent microsphere sets, we covalently coupled
T4-BSA and anti-TSH capture antibody to two
distinct microsphere sets. The anti-TSH monoclonal capture antibody
(100 µg), clone 204-12410 (OEM Concepts), and 25 µg of the
T4-BSA antigen (Fitzgerald Industries
International) were separately covalently attached to the carboxylate
groups
Acknowledgments
References
The following articles in journals at HighWire Press have cited this article:
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  K. Skogstrand, P. Thorsen, B. Norgaard-Pedersen, D. E. Schendel, L. C. Sorensen, and D. M. Hougaard Simultaneous Measurement of 25 Inflammatory Markers and Neurotrophins in Neonatal Dried Blood Spots by Immunoassay with xMAP Technology Clin. Chem., October 1, 2005; 51(10): 1854 - 1866. [Abstract] [Full Text] [PDF]
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S. Faucher, A. Martel, A. Sherring, T. Ding, L. Malloch, J. E. Kim, M. Bergeron, P. Sandstrom, and F. F. Mandy Protein Bead Array for the Detection of HIV-1 Antibodies from Fresh Plasma and Dried-Blood-Spot Specimens Clin. Chem., July 1, 2004; 50(7): 1250 - 1253. [Full Text] [PDF]
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D. H. Chace, T. A. Kalas, and E. W. Naylor Use of Tandem Mass Spectrometry for Multianalyte Screening of Dried Blood Specimens from Newborns Clin. Chem., November 1, 2003; 49(11): 1797 - 1817. [Abstract] [Full Text] [PDF]
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 R. E. Biagini, S. A. Schlottmann, D. L. Sammons, J. P. Smith, J. C. Snawder, C. A. F. Striley, B. A. MacKenzie, and D. N. Weissman Method for Simultaneous Measurement of Antibodies to 23 Pneumococcal Capsular Polysaccharides Clin. Vaccine Immunol., September 1, 2003; 10(5): 744 - 750. [Abstract] [Full Text] [PDF]
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Z. Lukacs, C. Mordac, A. Kohlschutter, and R. Kruithof Use of Microsphere Immunoassay for Simplified Multianalyte Screening of Thyrotropin and Thyroxine in Dried Blood Spots from Newborns Clin. Chem., February 1, 2003; 49(2): 335 - 336. [Full Text] [PDF]
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K. A. Pass Commentary on: Use of Microsphere Immunoassay for Simplified Multianalyte Screening of Thyrotropin and Thyroxine in Dried Blood Spots from Newborns Clin. Chem., February 1, 2003; 49(2): 336 - 336. [Full Text] [PDF]
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