- Blood spot
- Real-time PCR
- Neonatal screening
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.
Copyright © 2008 S. Karger AG, Basel
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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 firstname.lastname@example.org
Received: October 16, 2007
Accepted after revision: April 22, 2008
Published online: September 18, 2008
Number of Print Pages : 7
Number of Figures : 3, Number of Tables : 1, Number of References : 39
Neonatology (Fetal and Neonatal Research)
Vol. 95, No. 3, Year 2009 (Cover Date: March 2009)
Journal Editor: Halliday H.L. (Belfast), Speer C.P. (Würzburg)
ISSN: 1661–7800 (Print), eISSN: 1661–7819 (Online)
For additional information: http://www.karger.com/NEO
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