Absence of Cytomegalovirus in Glioblastoma and Other High-grade Gliomas by Real-time PCR, Immunohistochemistry, and In Situ Hybridization
Absence of Cytomegalovirus in Glioblastoma and Other High-grade Gliomas by Real-time PCR, Immunohistochemistry, and In Situ Hybridization
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DOI：
10.1158/1078-0432.ccr-16-1490
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发表时间：
2017-06-15
影响因子：
11.5
通讯作者：
Arav-Boger, Ravit
中科院分区：
文献类型：
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作者：
Holdhoff, Matthias;Guner, Gunes;Arav-Boger, Ravit
目的：高级别胶质瘤（HGG）/胶质母细胞瘤中巨细胞病毒（CMV）检测的报道一直存在争议。我们进行了一个全面的方法来确定CMV的存在或不存在的组织，血浆和血清中的HGG patients.Experimental Design：在一个回顾性的手臂，25个新鲜的冷冻组织胶质母细胞瘤患者进行了CMV实时PCR检测。通过IHC检测了70例HGG患者的组织微阵列，并通过IHC和显色原位杂交（CISH）检测了20例福尔马林固定石蜡包埋（FFPE）胶质母细胞瘤组织，靶向CMV编码的IE 1/2和pp 65。在一个前瞻性的手臂，18例新诊断的HGG提供组织和血液samples.Results：所有回顾性收集的组织均为CMV阴性的所有方法。在前瞻性队列中，18例新诊断的HGG患者在诊断时和随访期间提供了血液样本。在38份血浆标本中，基线时18份标本中有3份检出CMV DNA，20份随访标本中有1份检出CMV DNA。15例患者中8例（53%）血清CMVIgG阳性。在前瞻性研究组中检测的FFPE样本中，所有样本经IHC、CISH和PCR检测均为CMV阴性。利用6种高灵敏度检测试剂盒和3种正交技术，对多种标本和标本类型进行检测，我们的研究结果要求对从不同地理区域收集的样本进行多中心盲法分析，并商定研究设计和因果关系的确定，HGG/胶质母细胞瘤中缺乏CMV，以指导未来必要的抗病毒和/或基于CMV的治疗。(C)2016年AACR。
Purpose: Reports of cytomegalovirus (CMV) detection in high-grade gliomas (HGG)/glioblastoma have been conflicting. We undertook a comprehensive approach to determine the presence or absence of CMV in tissue, plasma, and serum of HGG patients.Experimental Design: In a retrospective arm, 25 fresh frozen tissues from glioblastoma patients were tested for CMV by real-time PCR. Tissue microarrays from 70 HGG patients were tested by IHC and 20 formalin-fixed paraffin-embedded (FFPE) glioblastoma tissues by IHC and chromogenic in situ hybridization (CISH), targeting CMV-encoded IE1/2 and pp65. In a prospective arm, 18 patients with newly diagnosed HGG provided tissue and blood samples.Results: All retrospectively collected tissues were negative for CMV by all methods. In the prospective cohort, 18 patients with newly diagnosed HGG provided blood samples at the time of diagnosis and during follow-up. Of 38 plasma specimens, CMV DNA was detected in 3 of 18 samples at baseline and 1 of 20 follow-up samples. Serum CMVIgG was positive in 8 of 15 (53%) of patients. Among the FFPE samples tested in the prospective arm, all were negative for CMV by IHC, CISH, and PCR.Conclusions: Utilizing 6 highly sensitive assays with three orthogonal technologies on multiple specimens and specimen types, no evidence for CMV in glioblastoma tissues was found. Our findings call for multicenter blinded analyses of samples collected from different geographical areas with agreed upon study designs and determination of causality or lack thereof of CMV in HGG/glioblastoma for future guidance on the necessary antiviral and/or CMV-based therapies. (C) 2016 AACR.