Comparison of 'real-time' and immunometric RT-PCR: RNA interference of reverse transcriptase-PCR.
We measured interleukin-6 (IL-6) mRNA in whole blood, using an immunometric reverse transcriptase-polymerase chain reaction (IRT-PCR) method and 'real-time' RT-PCR using the Roche LightCycler. Both methods showed similar precision, in terms of inter- and intra-assay variation, where a lower total RNA concentration was used at a relatively high ratio of (IL-6) mRNA to total RNA. However, the 'real-time' RT-PCR method showed a greater loss of accuracy at higher concentrations of total RNA.
Hoadley
Margaret E
ME
Injury Research, University of Manchester Division of Medicine and Neurosciences, Clinical Sciences Building, Hope Hospital, Salford, M6 8HD, UK. mhoadley@fsl.ho.man.ac.uk
Hopkins
Stephen J
SJ
D006801:Humans; D007118:Immunoassay / Q000379:methods; D015850:Interleukin-6 / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D034622:RNA Interference; D012333:RNA, Messenger / Q000097:blood*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D015850:Interleukin-6; D012333:RNA, Messenger