Comparing microarray versus RT-PCR assessment of renal allograft biopsies: Similar performance despite different dynamic ranges
Comparing microarray versus RT-PCR assessment of renal allograft biopsies: Similar performance despite different dynamic ranges
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DOI：
10.1111/j.1600-6143.2008.02199.x
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发表时间：
2008-05-01
影响因子：
8.8
通讯作者：
Halloran, P. F.
中科院分区：
文献类型：
--
作者：
Allanach, K.;Mengel, M.;Halloran, P. F.
在肾移植中，评估基因表达可以为组织病理学提供相关的诊断信息。结果可以表示为单个基因或基因集，代表基于发病机制的转录物集（PBT）：细胞毒性T细胞相关的、干扰素γ诱导的或减少的肾实质转录物。有两种技术平台可供使用：RT-PCR和微阵列。我们比较了RT-PCR，U133plus2.0微阵列和组织病理学在86个活检。我们比较了13个潜在的诊断基因作为RT-PCR探针，微阵列衍生的PBTs，“迷你”PBTs（3 - 5个转录本的小集合）和组织学分类器。大多数RT-PCR探针（10/13）与相应的微阵列探针组具有良好的相关性（r> 0.8）。这些探针包括FASLG和CD8B1微阵列探针组，它们不能在微阵列上执行，但通过RT-PCR可以检测到，这很可能是由于灵敏度的差异。总的来说，RT-PCR显示出更大的动态范围，检测正常肾脏的微小变化，但RT-PCR和微阵列在异常肾脏中得到了相似的结果。通过任一平台评估的个体转录物或微型PBTs与微阵列PBTs和组织学分类器彼此相关性良好。因此，微阵列和RT-PCR评估在评估移植炎症方面彼此和组织病理学高度一致，特别是当结果表示为PBTs或mini-PBTs时。两个平台的动态范围足以检测到排斥反应的相关变化。
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.