Real-time PCR HPV genotyping in fine needle aspirations of metastatic head and neck squamous cell carcinoma: Exposing the limitations of conventional p16 immunostaining
Real-time PCR HPV genotyping in fine needle aspirations of metastatic head and neck squamous cell carcinoma: Exposing the limitations of conventional p16 immunostaining
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DOI：
10.1016/j.oraloncology.2019.02.006
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发表时间：
2019-03-01
期刊：
影响因子：
4.8
通讯作者：
Westra, W. H.
中科院分区：
文献类型：
--
作者：
El-Salem, F.;Mansour, M.;Westra, W. H.
背景：鉴于hpv阳性的头颈部鳞状细胞癌（HPV-HNSCC）倾向于转移到颈部淋巴结，细针穿刺（FNA）在其初始检测中起着重要的诊断作用。事实上，即使没有明确的方法学指导方针和阈值标准，现在对FNAs的HPV检测也有坚定的承诺。一个特别的困难是p16染色的解释。设计：收集了210例疑似局部转移性HNSCC患者的数据，这些患者接受了FNA作为标准临床治疗的一部分。使用靶向高危型HPV L1的引物，在细胞块上用实时PCR进行初始HPV筛查。对hpv阳性病例进行额外的基因分型。将结果与p16染色和随后可用的切除进行比较。结果：207例样本中，175例（85%）为HPV阳性。HPV-16是最常见的基因型（90%）。hpv阳性病例中，原发部位为口咽部（n=154, 88.0%）、声门上喉（n=2, 1.1%）、鼻腔（n=1, 0.6%）、下咽（n=1, 0.6%）或未知（n=17, 9.7%）。与31例配对手术切除相比，所有病例的HPV状态一致（100%相关）。在142例与p16染色相匹配的hpv阳性病例中，p16染色报告为阳性（n= 85,60%）、局灶性（n= 27,19%）、阴性（n= 24,17%）或非贡献性（n= 6,4%）；只有33%达到HPV阳性的标准阈值（即70%）。结论：对于转移性HNSCC患者，FNAs的实时PCR可靠地反映HPV状态，优于传统的p16免疫染色。
Background: Given the propensity for HPV-positive head and neck squamous cell carcinoma (HPV-HNSCC) to metastasize to cervical lymph nodes, fine needle aspiration (FNA) plays an important diagnostic role in their initial detection. Indeed, there is now an unwavering commitment to HPV testing of FNAs even in the absence of clear methodologic guidelines and threshold criteria. A particular difficulty pertains to the interpretation of p16 staining.Design: Data was collected for 210 patients with suspected regionally metastatic HNSCC that had undergone FNA as part of standard clinical care. Initial HPV screening was performed on cell blocks with real-time PCR using primers targeting L1 of high-risk HPV types. Additional genotyping was performed on HPV-positive cases. The results were compared to p16 staining and subsequent excisions when available.Results: Of the 207 samples with sufficient DNA, 175 (85%) were HPV positive. HPV-16 was the most commonly detected genotype (90%). Of the HPV-positive cases, the primary site was the oropharynx (n=154, 88.0%), supraglottic larynx (n=2, 1.1%), nasal cavity (n=1, 0.6%), hypopharynx (n=1, 0.6%) or unknown (n=17, 9.7%). On comparison with 31 paired surgical excisions, HPV status was concordant in all cases (100% correlation). Of 142 HPV-positive cases with matching p16 stains, p16 staining was reported as positive (n=85, 60%), focal (n=27, 19%), negative (n=24, 17%) or non-contributory (n=6, 4%); and only 33% reached the standard threshold limit (i.e. 70%) for HPV positivity.Conclusion: For patients with metastatic HNSCC, real-time PCR of FNAs reliably reflects HPV status, and is superior to conventional p16 immunostaining.