A novel algorithm for reliable detection of human papillomavirus in paraffin embedded head and neck cancer specimen
A novel algorithm for reliable detection of human papillomavirus in paraffin embedded head and neck cancer specimen
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DOI：
10.1002/ijc.22980
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发表时间：
2007-12-01
影响因子：
6.4
通讯作者：
Brakenhoff, Ruud H.
中科院分区：
文献类型：
--
作者：
Smeets, Serge J.;Hesselink, Albertus T.;Brakenhoff, Ruud H.
人乳头瘤病毒16型（HPV 16）在头颈部鳞状细胞癌（HNSCC）的发展中起作用。然而，关于HPV在这种肿瘤类型中的真实影响存在不确定性，因为已经发表了相互矛盾的报告，患病率从0到100%。我们的目的是找到一种生物学上和临床上有意义的感染的检测算法，适用于冷冻和福尔马林固定石蜡包埋（FFPE）标本的高通量筛选。通过将冷冻活检中HPV E6癌基因表达的检测作为有意义的HPV感染的金标准，对48例HNSCC患者的FFPE肿瘤标本和血清进行了几种检测方法的价值评估。对FFPE组织样本进行了以下检测：HPV DNA通用引物（GP）5+/6+ PCR、病毒载量分析、HPV 16 DNA FISH检测、HPV 16 E6 mRNA RT-PCR、p16免疫染色，并对相应的血清样本进行了针对HPV 16蛋白L1、E6和E7的抗体检测。RT-PCR检测FFPE组织标本中E6的表达具有上级优势，但目前应用仍局限于HPV 16的检测。p16免疫组化和GP 5 +/6+ PCR对p16阳性病例的敏感性和特异性均为100%。我们表明，在FFPE HNSCC样本中，可以以标准和高通量方式更可靠地测量具有临床意义的病毒HPV感染，为预后和实验性疫苗接种研究铺平了道路，不仅涉及HNSCC，还可能涉及HPV参与的癌症类型，如阴茎癌和肛门癌。(C)2007 Wiley-Liss，Inc.
Human papillomavirus type 16 (HPV16) plays a role in the development of a subgroup of head and neck squamous cell carcinomas (HNSCC). However, uncertainty exists about the true impact of HPV in this tumor type as conflicting reports have been published with prevalence rates from 0 to 100%. We aimed to find a detection algorithm of a biologically and thus clinically meaningful infection, applicable for high-throughput screening of frozen and formalin-fixed paraffin embedded (FFPE) specimens. By considering detection of HPV E6 oncogene expression in frozen biopsies as gold standard for a meaningful HPV infection, the value of several assays was evaluated on FFPE tumor specimens and sera of 48 HNSCC patients. The following assays were evaluated on FFPE tissue samples: HPV DNA general primer (GP)5+/6+ PCR, viral load analysis, HPV16 DNA FISH detection, HPV16 E6 mRNA RT-PCR, p16 immunostaining, and on corresponding serum samples detection of antibodies against the HPV16 proteins L1, E6 and E7. Comparing single assays on FFPE tissue samples detection of E6 expression by RT-PCR was superior, but application remains at present limited to HPV16 detection. Most suitable algorithm with 100% sensitivity and specificity appeared p16 immunostaining followed by GP5+/6+ PCR on the p16-positive cases. We show that clinically meaningful viral HPV infections can be more reliably measured in FFPE HNSCC samples in a standard and high throughput manner, paving the way for prognostic and experimental vaccination studies, regarding not only HNSCC, but possibly also cancer types with HPV involvement in subgroups such as penile and anal cancer. (C) 2007 Wiley-Liss, Inc.