Archived unfrozen neonatal blood spots are amenable to quantitative gene expression analysis.
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.


OBJECTIVE
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.
Haak
Peterson T
PT
Laboratory of Microarray Technology, Van Andel Research Institute, Grand Rapids, Mich. 49503, USA.
Busik
Julia V
JV
Kort
Eric J
EJ
Tikhonenko
Maria
M
Paneth
Nigel
N
Resau
James H
JH
D001800:Blood Specimen Collection; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D015997:Neonatal Screening / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000097:blood*; D013997:Time Factors
D012333:RNA, Messenger
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