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Acquiring genome‐wide gene expression profiles in Guthrie card blood spots using microarrays
Author(s) -
Khoo Sok Kean,
Dykema Karl,
Vadlapatla Naga Manjari,
LaHaie David,
Valle Saul,
Satterthwaite David,
Ramirez Sara Angeline,
Carruthers Janell Alee,
Haak Peterson Todd,
Resau James Howard
Publication year - 2011
Publication title -
pathology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.73
H-Index - 74
eISSN - 1440-1827
pISSN - 1320-5463
DOI - 10.1111/j.1440-1827.2010.02611.x
Subject(s) - dna microarray , microarray , biology , nucleic acid , rna , genome , dried blood spot , gene expression , computational biology , complementary dna , gene , spots , human genome , gene expression profiling , microbiology and biotechnology , genetics , botany
Standard Guthrie cards have been widely used to collect blood samples from essentially all USA and Japanese neonates for newborn screening programs. Thus, archival blood spot samples are a unique and comprehensive resource for molecular pathology studies. However, the challenge in using these samples is the presumed low quantity and degraded quality of nucleic acids that can be isolated from these samples, particularly the RNA. Here, we report a new assay using Agilent 4x44K microarrays for acquiring genome‐wide gene expression profiles from blood spots on Guthrie cards. Due to the small amount of RNA obtained from each sample, major modifications, such as concentrating and amplifying the RNA and using a different labeling procedure, were performed. Approximately 9000 expressed genes can be detected after normalization of data, an increment of 260% in detection power compared with previously reported cDNA microarrays made in‐house with standard procedures. The correlation coefficients in technical and biological replicates were 0.92 and 0.85, respectively, confirming the reproducibility of this study. This new and comprehensive assay will add value to the utility of archival Guthrie cards (e.g. neonatal blood spot cards) and open new opportunities to molecular epidemiology, pathology, genomic, and diagnostic studies of perinatal diseases.