Comparing microarray versus RT-PCR assessment of renal allograft biopsies: Similar performance despite different dynamic ranges

被引:51
作者
Allanach, K. [1 ]
Mengel, M. [1 ]
Einecke, G. [1 ]
Sis, B. [1 ,2 ]
Hidalgo, L. G. [1 ]
Mueller, T. [1 ]
Halloran, P. F. [1 ]
机构
[1] Univ Alberta, Dept Med, Div Nephrol & Immunol, Alberta Transplant Appl Genom Ctr, Edmonton, AB, Canada
[2] Univ Alberta, Dept Lab Med & Pathol, Edmonton, AB, Canada
关键词
gene expression analysis; kidney transplantation; real time PCR;
D O I
10.1111/j.1600-6143.2008.02199.x
中图分类号
R61 [外科手术学];
学科分类号
摘要
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.
引用
收藏
页码:1006 / 1015
页数:10
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