Comparison of diagnostic methods for detection of BRAFV600E mutation in ameloblastoma

IF 2.7 3区医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE

Journal of Oral Pathology & Medicine Pub Date : 2024-01-07 DOI:10.1111/jop.13506

Arularasan Anbinselvam, Abdul-Warith O. Akinshipo, Akinyele O. Adisa, Olajumoke A. Effiom, Xinhe Zhu, Kehinde E. Adebiyi, Godwin T. Arotiba, Sunday O. Akintoye

{"title":"Comparison of diagnostic methods for detection of BRAFV600E mutation in ameloblastoma","authors":"Arularasan Anbinselvam, Abdul-Warith O. Akinshipo, Akinyele O. Adisa, Olajumoke A. Effiom, Xinhe Zhu, Kehinde E. Adebiyi, Godwin T. Arotiba, Sunday O. Akintoye","doi":"10.1111/jop.13506","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Background</h3>\n \n <p>Ameloblastoma is an aggressively growing, highly recurrent odontogenic jaw tumor. Its association with BRAFV600E mutation is an indication for BRAFV00E-inhibitor therapy The study objective was to identify a sensitive low-cost test for BRAFV600E-positive ameloblastoma. We hypothesized that immunohistochemical staining of formalin-fixed paraffin-embedded tissues for BRAFV600E mutation is a low-cost surrogate for BRAFV600E gene sequencing when laboratory resources are inadequate for molecular testing.</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Tissues from 40 ameloblastoma samples were retrieved from either formalin-fixed paraffin-embedded blocks, RNAlater™ stabilization solution or samples inadvertently pre-fixed in formalin before transfer to RNAlater™. BRAFV600E mutation was assessed by Direct Sanger sequencing, Amplification Refractory Mutation System-PCR and immunohistochemistry (IHC).</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>BRAFV600E mutation was detected by IHC, Amplification Refractory Mutation System-PCR and Direct Sanger sequencing in 93.33%, 52.5% and 30% of samples respectively. Considering Direct Sanger sequencing as standard BRAFV600E detection method, there was significant difference between the three detection methods (𝜒<sup>2</sup> (2) = 31.34, <i>p</i> < 0.0001). Sensitivity and specificity of IHC were 0.8 (95% CI: 0.64–0.90) and 0.9 (95% CI: 0.75–0.99) respectively, while positive predictive value and negative predictive value (NPV) were 0.9 and 0.8 (Fischer's test, <i>p</i> < 0.0001) respectively. Sensitivity and specificity of Amplification Refractory Mutation System-PCR detection method were 0.7 (95% CI: 0.53–0.80) and 0.9 (95% CI = 0.67–0.98) respectively, while PPV and NPV were 0.9 and 0.6 respectively (Fischer's test, <i>p</i> < 0.0001).</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>Low-cost and less vulnerability of IHC to tissue quality make it a viable surrogate test for BRAFV600E detection in ameloblastoma. Sequential dual IHC and molecular testing for BRAFV600E will reduce equivocal results that could exclude some patients from BRAFV600E-inhibitor therapies.</p>\n </section>\n </div>","PeriodicalId":16588,"journal":{"name":"Journal of Oral Pathology & Medicine","volume":"53 1","pages":"79-87"},"PeriodicalIF":2.7000,"publicationDate":"2024-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Oral Pathology & Medicine","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1111/jop.13506","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"DENTISTRY, ORAL SURGERY & MEDICINE","Score":null,"Total":0}

引用次数: 0

Abstract

Background

Ameloblastoma is an aggressively growing, highly recurrent odontogenic jaw tumor. Its association with BRAFV600E mutation is an indication for BRAFV00E-inhibitor therapy The study objective was to identify a sensitive low-cost test for BRAFV600E-positive ameloblastoma. We hypothesized that immunohistochemical staining of formalin-fixed paraffin-embedded tissues for BRAFV600E mutation is a low-cost surrogate for BRAFV600E gene sequencing when laboratory resources are inadequate for molecular testing.

Methods

Tissues from 40 ameloblastoma samples were retrieved from either formalin-fixed paraffin-embedded blocks, RNAlater™ stabilization solution or samples inadvertently pre-fixed in formalin before transfer to RNAlater™. BRAFV600E mutation was assessed by Direct Sanger sequencing, Amplification Refractory Mutation System-PCR and immunohistochemistry (IHC).

Results

BRAFV600E mutation was detected by IHC, Amplification Refractory Mutation System-PCR and Direct Sanger sequencing in 93.33%, 52.5% and 30% of samples respectively. Considering Direct Sanger sequencing as standard BRAFV600E detection method, there was significant difference between the three detection methods (𝜒² (2) = 31.34, p < 0.0001). Sensitivity and specificity of IHC were 0.8 (95% CI: 0.64–0.90) and 0.9 (95% CI: 0.75–0.99) respectively, while positive predictive value and negative predictive value (NPV) were 0.9 and 0.8 (Fischer's test, p < 0.0001) respectively. Sensitivity and specificity of Amplification Refractory Mutation System-PCR detection method were 0.7 (95% CI: 0.53–0.80) and 0.9 (95% CI = 0.67–0.98) respectively, while PPV and NPV were 0.9 and 0.6 respectively (Fischer's test, p < 0.0001).

Conclusion

Low-cost and less vulnerability of IHC to tissue quality make it a viable surrogate test for BRAFV600E detection in ameloblastoma. Sequential dual IHC and molecular testing for BRAFV600E will reduce equivocal results that could exclude some patients from BRAFV600E-inhibitor therapies.

查看原文本刊更多论文

检测母细胞瘤中 BRAFV600E 突变的诊断方法比较。

背景：釉母细胞瘤是一种生长迅速、高度复发的牙源性颌骨肿瘤。该研究的目的是找出一种灵敏、低成本的检测方法来检测 BRAFV600E 阳性的釉母细胞瘤。我们假设，当实验室资源不足以进行分子检测时，对福尔马林固定石蜡包埋组织进行免疫组化染色以检测 BRAFV600E 突变是一种低成本的 BRAFV600E 基因测序替代方法：从福尔马林固定的石蜡包埋块、RNAlater™稳定溶液或转移至RNAlater™前不慎预固定在福尔马林中的样本中提取40个骨髓母细胞瘤样本的组织。通过直接桑格测序、扩增难治性突变系统-PCR和免疫组织化学（IHC）评估BRAFV600E突变：结果：93.33%、52.5%和30%的样本分别通过IHC、扩增难治性突变系统-PCR和直接桑格测序检测到BRAFV600E突变。将直接桑格测序法作为标准的 BRAFV600E 检测方法，三种检测方法之间存在显著差异（𝜒2 (2) = 31.34，p 结论：直接桑格测序法检测 BRAFV600E 基因突变的成本低，且不容易被检测到：IHC成本低且不易受组织质量的影响，因此是检测母细胞瘤中BRAFV600E的可行替代检测方法。对BRAFV600E进行连续的双重IHC和分子检测将减少可能将某些患者排除在BRAFV600E抑制剂疗法之外的不明确结果。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Journal of Oral Pathology & Medicine 医学-病理学

CiteScore

5.90

自引率

6.10%

发文量

121

审稿时长

4-8 weeks

期刊介绍： The aim of the Journal of Oral Pathology & Medicine is to publish manuscripts of high scientific quality representing original clinical, diagnostic or experimental work in oral pathology and oral medicine. Papers advancing the science or practice of these disciplines will be welcomed, especially those which bring new knowledge and observations from the application of techniques within the spheres of light and electron microscopy, tissue and organ culture, immunology, histochemistry and immunocytochemistry, microbiology, genetics and biochemistry.