title: Evaluation of two commercial global miRNA expression profiling platforms for detection of less abundant miRNAs.
abstract: BACKGROUND
  microRNAs (miRNA) are short, endogenous transcripts that negatively regulate the expression of specific mRNA targets. miRNAs are found both in tissues and body fluids such as plasma. A major perspective for the use of miRNAs in the clinical setting is as diagnostic plasma markers for neoplasia. While miRNAs are abundant in tissues, they are often scarce in plasma. For quantification of miRNA in plasma it is therefore of importance to use a platform with high sensitivity and linear performance in the low concentration range. This motivated us to evaluate the performance of three commonly used commercial miRNA quantification platforms: GeneChip miRNA 2.0 Array, miRCURY Ready-to-Use PCR, Human panel I+II V1.M, and TaqMan Human MicroRNA Array v3.0.
  
  
  RESULTS
  Using synthetic miRNA samples and plasma RNA samples spiked with different ratios of 174 synthetic miRNAs we assessed the performance characteristics reproducibility, recovery, specificity, sensitivity and linearity. It was found that while the qRT-PCR based platforms were sufficiently sensitive to reproducibly detect miRNAs at the abundance levels found in human plasma, the array based platform was not. At high miRNA levels both qRT-PCR based platforms performed well in terms of specificity, reproducibility and recovery. At low miRNA levels, as in plasma, the miRCURY platform showed better sensitivity and linearity than the TaqMan platform.
  
  
  CONCLUSIONS
  For profiling clinical samples with low miRNA abundance, such as plasma samples, the miRCURY platform with its better sensitivity and linearity would probably be superior.
authors.affiliation: Department of Molecular Medicine (MOMA), Aarhus University Hospital-Skejby, DK-8200 Aarhus N, Denmark.
authors.forename: Steffen G
authors.initials: SG
authors.lastname: Jensen
authors.forename: Philippe
authors.initials: P
authors.lastname: Lamy
authors.forename: Mads H
authors.initials: MH
authors.lastname: Rasmussen
authors.forename: Marie S
authors.initials: MS
authors.lastname: Ostenfeld
authors.forename: Lars
authors.initials: L
authors.lastname: Dyrskjøt
authors.forename: Torben F
authors.initials: TF
authors.lastname: Orntoft
authors.forename: Claus L
authors.initials: CL
authors.lastname: Andersen
mesh_terms: D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000302:isolation & purification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D035683:MicroRNAs