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

Am J Transplant. 2008 May;8(5):1006-15. doi: 10.1111/j.1600-6143.2008.02199.x.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biopsy
  • Canada
  • Ethnicity / genetics
  • Humans
  • Kidney Transplantation / immunology
  • Kidney Transplantation / pathology*
  • Oligonucleotide Array Sequence Analysis*
  • RNA / genetics
  • RNA / isolation & purification
  • Racial Groups / genetics
  • Reverse Transcriptase Polymerase Chain Reaction*
  • T-Lymphocytes, Cytotoxic / immunology
  • Transcription, Genetic
  • Transplantation, Homologous / immunology
  • Transplantation, Homologous / pathology

Substances

  • RNA