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Archived Unfrozen Neonatal Blood Spots Are Amenable to Quantitative Gene Expression AnalysisHaak P.T.a, c, e · Busik J.V.f · Kort E.J.b · Tikhonenko M.f · Paneth N.c, d · Resau J.H.a, b
Laboratories of aMicroarray Technology and bMolecular Epidemiology, Van Andel Research Institute, Grand Rapids, Mich., Departments of cEpidemiology and dPediatrics and Human Development, College of Human Medicine, eCollege of Osteopathic Medicine, and fDepartment of Physiology, Michigan State University, East Lansing, Mich., USA Corresponding Author
Dr. James H. Resau
Van Andel Research Institute
333 Bostwick Ave., NE
Grand Rapids, MI 49503 (USA)
Tel. +1 616 234 5288, Fax +1 616 234 5289, E-Mail email@example.com
Background: State laws in the USA mandate that blood be drawn from all newborn infants to screen for health-threatening conditions. These screening assays consume only a small portion of the blood samples, which are collected on filter paper (‘Guthrie’) cards. Many states archive unused blood spots, often in unrefrigerated storage. Objectives: While individual RNA transcripts have been identified from archived neonatal blood spots, no study to date has performed quantitative analysis of archived blood spot RNA. Methods: We demonstrate that RNA can be isolated and amplified from newborn blood spots stored unfrozen for as long as 9 years, and can be analyzed by microarray and qPCR. Results: Microarray assays of archived neonatal blood spots consistently detected 3,000–4,000 expressed genes with correlations of 0.90 between replicates. Blood spot mRNA is amenable to qPCR and we detected biologically relevant expression levels of housekeeping and immune-mediating genes. Conclusions: These experiments demonstrate the feasibility of using blood spots as a source of RNA which can be analyzed using quantitative microarray and qPCR assays. The application of these methods to the analysis of widely collected biological specimens may be a valuable resource for the study of perinatal determinants of disease development.
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