Molecular Diagnosis of Ewing's Sarcoma/Primitive Neuroectodermal Tumor in Formalin-Fixed Paraffin-Embedded Tissues by RT-PCR and Fluorescence In Situ Hybridization
Molecular Diagnosis of Ewing's Sarcoma/Primitive Neuroectodermal Tumor in Formalin-Fixed Paraffin-Embedded Tissues by RT-PCR and Fluorescence In Situ Hybridization
复制标题
RT-PCR 和荧光原位杂交对福尔马林固定石蜡包埋组织中尤文氏肉瘤/原始神经外胚层肿瘤的分子诊断
DOI：
10.1097/01.pdm.0000140192.27878.97
复制
发表时间：
2005
影响因子：
--
通讯作者：
R. Lloyd
中科院分区：
文献类型：
--
作者：
X. Qian;Long Jin;B. Shearer;R. Ketterling;S. Jalal;R. Lloyd
最近的研究表明，福尔马林固定石蜡包埋(FFPE)组织可用于荧光原位杂交(FISH)和RT-PCR的分子分析。我们对18例ES/PNET的t(11；22)(q24；q12)和t(21；22)(q22；q12)融合转录本进行RT-PCR分析，并用双色融合探针间期FISH分析EWS易位，以直接比较两种方法。RT-PCR检测到13例(72%)EWS-FLI-1融合(I型10例，II型3例)和2例(11%)EWS-ERG融合。有3例因看家基因磷酸甘油酸激酶(内对照)未扩增而不能进行评估。FISH诊断15例(83%)。RT-PCR法与FISH法有3例不符合，符合率为83%。使用RT-PCR和FISH相结合的方法，结果是互补的。RT-PCR分析的一个优点是可以特异性地确定EWS易位的亚型(I型、II型和ERG)。这些发现表明，由于FFPE组织分子分析的困难和局限性，RT-PCR和FISH相结合可能是一种更好的方法来提高对保存不佳的核酸的FFPE组织中ES/PNET易位的检测的敏感性和准确性。
Recent studies have suggested that formalin-fixed paraffin-embedded (FFPE) tissues can be used for molecular analyses by fluorescence in situ hybridization (FISH) and RT-PCR. We analyzed 18 cases of ES/PNET for the t(11;22)(q24;q12) and t(21;22)(q22;q12) fusion transcripts by RT-PCR and analyzed for EWS translocation by interphase FISH with a dual color fusion probe to compare these two approaches directly. RT-PCR detected 13 (72%) EWS-FLI-1 fusions (type I=10, type II=3) and 2 (11%) EWS-ERG fusions. Three cases could not be evaluated because the housekeeping gene phosphoglycerate kinase (internal mRNA control) was not amplified. FISH was diagnostic in 15 of 18 cases (83%). There were three discordant cases between RT-PCR and FISH (concordance of 83%). Using a combination of RT-PCR and FISH, the results were complementary. One advantage of RT-PCR analysis was that subtypes of EWS translocation could be determined specifically (type I, type II and ERG). These findings indicate that because of the difficulties and limitations associated with the molecular analysis of FFPE tissues, a combination of RT-PCR and FISH may be a better approach to enhance the sensitivity and accuracy of detecting ES/PNET translocations in FFPE tissues with suboptimally preserved nucleic acids.
影响因子：
45.3
作者：
de Alava, E;Kawai, A;Ladanyi, M
通讯作者：
Ladanyi, M