Validation of methods for oropharyngeal cancer HPV status determination in US cooperative group trials.
Validation of methods for oropharyngeal cancer HPV status determination in US cooperative group trials.
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DOI：
10.1097/pas.0b013e318253a2d1
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发表时间：
2012-07
期刊：
影响因子：
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通讯作者：
Gillison ML
中科院分区：
文献类型：
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作者：
Jordan RC;Lingen MW;Perez-Ordonez B;He X;Pickard R;Koluder M;Jiang B;Wakely P;Xiao W;Gillison ML
肿瘤HPV状态是口咽癌的预后因素，但分类方法尚未标准化。在这里，我们验证了美国合作组试验中使用的HPV分类方法。从2000-2009年诊断的240例石蜡包埋口咽癌中纯化的肿瘤DNA和RNA，如果通过定量PCR对DNA和mRNA对照呈阳性，则被评为可评价。通过一致性PCR检测肿瘤中的18种高危型HPV，然后通过定量逆转录酶PCR进行HR-HPV E6/7癌基因表达分析。以HR-HPV E6/7癌基因表达为对照，评价免疫组化（IHC）检测p16表达和原位杂交（ISH）检测HPV 16的敏感性（S）、特异性（SP）、阳性预测值（PPV）和阴性预测值（NPV）。三位病理学家之间的评价者一致性通过kappa统计进行评价。在235例可评估的肿瘤中，158例（6/7%，95%CI 61.2-73.3）HR-HPV E6/7癌基因表达阳性[HPV 16型（92%）、18型（3%）、33型（3%）、35型（1%）或58型（1%）]。p16 IHC检测HR-HPV癌基因表达的敏感性高[S 96.8%，SP 83.8%，PPV 92.7%，NPV 92.5%]，而HPV 16 ISH检测HR-HPV癌基因表达的特异性高[S 88.0%，SP 94.7%，PPV 97.2%，NPV 78.9%]。p16（kappa=0.95-0.98）和HPV 16 ISH（kappa 0.83-0.91）的评定者间一致性极佳。受试者工作曲线分析确定了p16强度评分和肿瘤染色百分比的叉积，以最佳区分HR-HPV E6/7阳性和阴性肿瘤。p16 IHC和HPV 16 ISH检测具有良好的性能，分别具有高灵敏度和特异性。适当的检测选择取决于假阳性或阴性检测的临床意义。
Tumor HPV status is a prognostic factor for oropharyngeal cancer, but classification methods are not standardized. Here we validate HPV classification methods used in United States cooperative group trials. Tumor DNA and RNA purified from 240 paraffin-embedded oropharyngeal cancers diagnosed from 2000–2009 were scored as evaluable if positive for DNA and mRNA controls by quantitative-PCR. Eighteen high-risk (HR)-HPV types were detected in tumors by consensus PCR followed by HR-HPV E6/7 oncogene expression analysis by quantitative reverse-transcriptase PCR. Sensitivity (S), specificity (SP), positive (PPV) and negative predictive value (NPV) of p16 expression detected by immunohistochemistry (IHC) and HPV16 detected by in situ hybridization (ISH) were evaluated in comparison to HR-HPV E6/7 oncogene expression. Inter-rater agreement among three pathologists was evaluated by kappa statistic. Of 235 evaluable tumors, 158 (67%, 95%CI 61.2–73.3) were positive for HR-HPV E6/7 oncogene expression [HPV type 16 (92%), 18 (3%), 33 (3%), 35 (1%) or 58 (1%)]. p16 IHC had high sensitivity [S 96.8%, SP 83.8%, PPV 92.7%, NPV 92.5%] whereas HPV16 ISH had high specificity [S 88.0%, SP 94.7%, PPV 97.2%, NPV 78.9%] for HR-HPV oncogene expression. Inter-rater agreement was excellent for p16 (kappa=0.95–0.98) and HPV16 ISH (kappa 0.83–0.91). Receiver-operating-curve analysis determined the cross-product of p16 intensity score and percent tumor staining to optimally discriminate HR-HPV E6/7-positive and negative tumors. p16 IHC and HPV16 ISH assays have excellent performance, with high sensitivity and specificity, respectively. Appropriate assay choice depends upon clinical implications of a false-positive or negative test.