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Molecular & Cellular Proteomics 7:1863-1875, 2008.
© 2008 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

Identification of Diagnostic Biomarkers for Infection in Premature Neonates^*

Stephen F. Kingsmore^,, Neil Kennedy^¶, Henry L. Halliday^¶, Jennifer C. Van Velkinburgh, Shengiang Zhong^||, Vanessa Gabriel^**, Judith Grant^**, William D. Beavis^||, Velizar T. Tchernev^,, Lorah Perlee, Serguei Lejnine, Brian Grimwade, Martin Sorette and J. David M. Edgar^¶¶

From the National Center for Genome Resources, Santa Fe, New Mexico, 87505, ^¶ Royal-Jubilee Maternity Service and ^¶¶ Regional Immunology Service, Royal Hospitals Trust, Belfast, Northern Ireland BT12 6BB, United Kingdom, ^|| Agronomy Department, Iowa State University, Ames, Iowa 50011, ^** Department of Neonatal Medicine, Queen's Medical Centre, Nottingham NG7 2UH, United Kingdom, and Molecular Staging, Inc., New Haven, Connecticut 06511

Infection is a leading cause of neonatal morbidity and mortality worldwide. Premature neonates are particularly susceptible to infection because of physiologic immaturity, comorbidity, and extraneous medical interventions. Additionally premature infants are at higher risk of progression to sepsis or severe sepsis, adverse outcomes, and antimicrobial toxicity. Currently initial diagnosis is based upon clinical suspicion accompanied by nonspecific clinical signs and is confirmed upon positive microbiologic culture results several days after institution of empiric therapy. There exists a significant need for rapid, objective, in vitro tests for diagnosis of infection in neonates who are experiencing clinical instability. We used immunoassays multiplexed on microarrays to identify differentially expressed serum proteins in clinically infected and non-infected neonates. Immunoassay arrays were effective for measurement of more than 100 cytokines in small volumes of serum available from neonates. Our analyses revealed significant alterations in levels of eight serum proteins in infected neonates that are associated with inflammation, coagulation, and fibrinolysis. Specifically P- and E-selectins, interleukin 2 soluble receptor , interleukin 18, neutrophil elastase, urokinase plasminogen activator and its cognate receptor, and C-reactive protein were observed at statistically significant increased levels. Multivariate classifiers based on combinations of serum analytes exhibited better diagnostic specificity and sensitivity than single analytes. Multiplexed immunoassays of serum cytokines may have clinical utility as an adjunct for rapid diagnosis of infection and differentiation of etiologic agent in neonates with clinical decompensation.

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