Abstract：
BACKGROUND: The objective of this study is to evaluate the diagnostic accuracy of plasma-based liquid biopsy for the detection of the BRAF V600E mutation in circulating cell-free DNA from patients with ameloblastoma.
METHODS: This is a prospective diagnostic accuracy study conducted based on the Standards for Reporting Diagnostic Accuracy recommendations. The index test was the plasma-based liquid biopsy, whereas the reference standard was the conventional tissue biopsy. The target condition was the detection of BRAF V600E mutation. The study population consisted of individuals with ameloblastoma recruited from three tertiary hospitals from Brazil. A negative control group composed of three individuals with confirmed wild-type BRAF lesions were included. The participants underwent plasma circulating cell-free DNA and tumor tissue DNA isolation, and both were submitted to using competitive allele-specific TaqMan™ real-time polymerase chain reaction technology mutation detection assays. Sensitivity and specificity measures and positive and negative predictive values were calculated.
RESULTS: Twelve patients with conventional ameloblastoma were included. BRAF V600E mutation was detected in 11/12 (91.66%) ameloblastoma tissue samples. However, the mutation was not detected in any of the plasma-based liquid biopsy circulating cell-free DNA samples in both ameloblastomas and negative control group. The sensitivity and specificity of plasma-based liquid biopsy for the detection of the BRAF V600E mutation in circulating cell-free DNA was 0.0 and 1.0, respectively. The agreement between index test and reference standard results was 26.66%.
CONCLUSIONS: Plasma-based liquid biopsy does not seem to be an accurate method for the detection of the BRAF V600E mutation in circulating circulating cell-free DNA from patients with ameloblastoma, regardless of tumor size, anatomic location, recurrence status, and other clinicopathological features.
METHODS: This is a prospective diagnostic accuracy study conducted based on the Standards for Reporting Diagnostic Accuracy recommendations. The index test was the plasma-based liquid biopsy, whereas the reference standard was the conventional tissue biopsy. The target condition was the detection of BRAF V600E mutation. The study population consisted of individuals with ameloblastoma recruited from three tertiary hospitals from Brazil. A negative control group composed of three individuals with confirmed wild-type BRAF lesions were included. The participants underwent plasma circulating cell-free DNA and tumor tissue DNA isolation, and both were submitted to using competitive allele-specific TaqMan™ real-time polymerase chain reaction technology mutation detection assays. Sensitivity and specificity measures and positive and negative predictive values were calculated.
RESULTS: Twelve patients with conventional ameloblastoma were included. BRAF V600E mutation was detected in 11/12 (91.66%) ameloblastoma tissue samples. However, the mutation was not detected in any of the plasma-based liquid biopsy circulating cell-free DNA samples in both ameloblastomas and negative control group. The sensitivity and specificity of plasma-based liquid biopsy for the detection of the BRAF V600E mutation in circulating cell-free DNA was 0.0 and 1.0, respectively. The agreement between index test and reference standard results was 26.66%.
CONCLUSIONS: Plasma-based liquid biopsy does not seem to be an accurate method for the detection of the BRAF V600E mutation in circulating circulating cell-free DNA from patients with ameloblastoma, regardless of tumor size, anatomic location, recurrence status, and other clinicopathological features.
摘要：
背景：本研究的目的是评估基于血浆的液体活检在成釉细胞瘤患者循环无细胞DNA中检测BRAFV600E突变的诊断准确性。
方法：这是一项基于诊断准确性报告标准建议的前瞻性诊断准确性研究。指数测试是基于血浆的液体活检，而参考标准是常规组织活检。目标条件为检测BRAFV600E突变。研究人群包括从巴西三家三级医院招募的成釉细胞瘤个体。包括由具有证实的野生型BRAF病变的三个个体组成的阴性对照组。参与者接受了血浆循环无细胞DNA和肿瘤组织DNA分离，并且两者都提交使用竞争性等位基因特异性TaqMan™实时聚合酶链反应技术突变检测测定。计算敏感性和特异性测量以及阳性和阴性预测值。
结果：纳入12例常规成釉细胞瘤患者。在11/12（91.66%）成釉细胞瘤组织样本中检测到BRAFV600E突变。然而,在成釉细胞瘤和阴性对照组的任何基于血浆的液体活检循环无细胞DNA样本中均未检测到该突变.基于血浆的液体活检检测循环无细胞DNA中BRAFV600E突变的敏感性和特异性分别为0.0和1.0。指标测试与参考标准结果的一致性为26.66%。
结论：基于血浆的液体活检似乎不是检测成釉细胞瘤患者循环循环循环无细胞DNA中BRAFV600E突变的准确方法，不管肿瘤大小，解剖位置,复发状态，和其他临床病理特征。
方法：这是一项基于诊断准确性报告标准建议的前瞻性诊断准确性研究。指数测试是基于血浆的液体活检，而参考标准是常规组织活检。目标条件为检测BRAFV600E突变。研究人群包括从巴西三家三级医院招募的成釉细胞瘤个体。包括由具有证实的野生型BRAF病变的三个个体组成的阴性对照组。参与者接受了血浆循环无细胞DNA和肿瘤组织DNA分离，并且两者都提交使用竞争性等位基因特异性TaqMan™实时聚合酶链反应技术突变检测测定。计算敏感性和特异性测量以及阳性和阴性预测值。
结果：纳入12例常规成釉细胞瘤患者。在11/12（91.66%）成釉细胞瘤组织样本中检测到BRAFV600E突变。然而,在成釉细胞瘤和阴性对照组的任何基于血浆的液体活检循环无细胞DNA样本中均未检测到该突变.基于血浆的液体活检检测循环无细胞DNA中BRAFV600E突变的敏感性和特异性分别为0.0和1.0。指标测试与参考标准结果的一致性为26.66%。
结论：基于血浆的液体活检似乎不是检测成釉细胞瘤患者循环循环循环无细胞DNA中BRAFV600E突变的准确方法，不管肿瘤大小，解剖位置,复发状态，和其他临床病理特征。
