Head and Neck Cancer Biobank, Guy's and St Thomas' NHS Foundation Trust, London, UK.
Head and Neck Pathology, Guy's and St Thomas' NHS Foundation Trust, London, UK.
J Oral Pathol Med. 2021 Jan;50(1):68-75. doi: 10.1111/jop.13103. Epub 2020 Sep 1.
The current diagnostic standard for detection of high-risk human papillomavirus (HPV) in oropharyngeal squamous cell carcinoma is via a two-stage algorithm, namely p16 immunohistochemistry followed by HPV DNA in situ hybridization in p16 positive cases. This study evaluated the feasibility of automated RNA in situ hybridization on a clinical platform as a single-step alternative to the two-stage algorithm within a routine diagnostic histopathology setting.
Thirty-eight cases positive for both p16 and DNA in situ hybridization, 42 p16 negative cases and 20 cases positive for p16 but negative for DNA in situ hybridization were randomly selected. High-risk HPV RNA in situ hybridization was undertaken on all cases on an automated clinical platform. Manufacturer-recommended and on-slide additional p16/HPV positive and negative controls were used. Test quality assurance and diagnostic RNA in situ hybridization were independently assessed by two observers. A consensus diagnosis was reached in the presence of a third observer on discordant cases. All RNA in situ hybridization results were then correlated against p16 and DNA ISH status.
Inter-slide RNA in situ hybridization staining variation was observed in control sections. RNA in situ hybridization demonstrated a high inter-observer agreement rate (κ = .897, P < .001). Following consensus review, there was full concordance between RNA in situ hybridization and the current standard.
Human papillomavirus testing by standalone automated RNA in situ hybridization on a clinical diagnostic platform currently available in routine diagnostic histopathology laboratories is a feasible alternative to the two-step algorithm of p16 and DNA in situ hybridization. Control tissue staining procedures need to be adapted to achieve the most accurate results.
目前，检测口咽鳞状细胞癌高危型人乳头瘤病毒（HPV）的诊断标准是采用两阶段算法，即 p16 免疫组化，然后在 p16 阳性病例中进行 HPV DNA 原位杂交。本研究评估了在常规诊断组织病理学环境中，自动化 RNA 原位杂交在临床平台上作为两步算法（p16 和 DNA 原位杂交）的替代方案的可行性。
随机选择 38 例 p16 和 DNA 原位杂交均阳性、42 例 p16 阴性和 20 例 p16 阳性但 DNA 原位杂交阴性的病例。所有病例均在自动化临床平台上进行高危型 HPV RNA 原位杂交。使用制造商推荐的和载玻片上的额外 p16/HPV 阳性和阴性对照。由两名观察者独立进行测试质量保证和诊断 RNA 原位杂交评估。在存在分歧的病例时，由第三位观察者达成共识诊断。然后将所有 RNA 原位杂交结果与 p16 和 DNA ISH 状态相关联。
在对照切片中观察到载玻片之间的 RNA 原位杂交染色变化。RNA 原位杂交显示出高的观察者间一致性率（κ=0.897，P<.001）。经过共识审查，RNA 原位杂交与当前标准之间完全一致。
在常规诊断组织病理学实验室中，目前可在临床诊断平台上进行独立的自动化 RNA 原位杂交，是 p16 和 DNA 原位杂交两步算法的可行替代方案。需要调整对照组织染色程序以获得最准确的结果。
