Comparing microarray versus RT-PCR assessment of renal allograft biopsies: similar performance despite different dynamic ranges.
In renal allografts, assessing gene expression can add relevant diagnostic information to histopathology. Results can be expressed as single genes or gene sets, representing pathogenesis-based transcript sets (PBTs): cytotoxic T-cell-associated, interferon gamma- induced or decreased kidney parenchymal transcripts. Two technology platforms are available: RT-PCR and microarrays. We compared RT-PCR, U133plus2.0 microarrays and histopathology in 86 biopsies. We compared 13 potentially diagnostic genes as RT-PCR probes to microarray-derived PBTs, 'mini'-PBTs (small sets of 3-5 transcripts) and a histology classifier. Most RT-PCR probes (10/13) correlated well with the corresponding microarray probe sets (r > 0.8). Exceptions included FASLG and CD8B1 microarray probe sets, which were not performing on microarrays but were detectable by RT-PCR most likely due to differences in sensitivity. In general, RT-PCR showed greater dynamic range, detecting small changes in normal kidneys, but RT-PCR and microarrays gave similar results in abnormal kidneys. Individual transcripts or mini-PBTs assessed by either platform correlated well with one another, with microarray PBTs and the histology classifier. Thus, microarrays and RT-PCR assessments agree strongly with one another and histopathology in assessing transplant inflammation, particularly, when results are expressed as PBTs or mini-PBTs. The dynamic range of both platforms was sufficient to detect the relevant changes in rejection.
Allanach
K
K
Department of Medicine, Division of Nephrology & Immunology, Alberta Transplant Applied Genomics Centre, University of Alberta, Edmonton, Canada.
Mengel
M
M
Einecke
G
G
Sis
B
B
Hidalgo
L G
LG
Mueller
T
T
Halloran
P F
PF
D001706:Biopsy; D002170:Canada; D005006:Ethnicity / Q000235:genetics; D006801:Humans; D016030:Kidney Transplantation / Q000276:immunology / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D044469:Racial Groups / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014158:Transcription, Genetic; D014184:Transplantation, Homologous / Q000276:immunology / Q000473:pathology
D012313:RNA